SCCS Notes of Guidance for the testing of cosmetic ingredients and

SCCS/1628/21

Scientific Committee on Consumer Safety

SCCS

THE SCCS NOTES OF GUIDANCE FOR THE TESTING OF

COSMETIC INGREDIENTS AND THEIR SAFETY

EVALUATION

REVISION

The SCCS adopted this guidance document

at its plenary meeting on 30-31 March 2021

 
 
 
 
 
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TABLE OF CONTENTS 
 
ACKNOWLEDGMENTS ................................................................................................ 1 
Main changes in 11th REVISION of the SCCS Notes of Guidance (NoG) ............................ 4 
1.  INTRODUCTION ................................................................................................... 5 
2.  THE SCIENTIFIC COMMITTEE ON CONSUMER SAFETY, SCCS..................................... 7 
2-1  BACKGROUND ............................................................................................... 7 
2-2  MANDATE ...................................................................................................... 7 
2-3  RULES OF PROCEDURE ................................................................................... 7 
2-4  OPINIONS ..................................................................................................... 7 
2-4.1  The "Notes of Guidance" ............................................................................. 7 
2-4.2  SCCS Cosmetic ingredient dossiers .............................................................. 8 
2-4.3  Specific issues taken up in NoG ................................................................... 9 
3.  SAFETY EVALUATION OF COSMETIC INGREDIENTS ................................................ 10 
3-1  SAFETY EVALUATION OF COSMETIC INGREDIENTS AS APPLIED BY THE SCCS ..... 10 
3-2   CHEMICAL AND PHYSICAL SPECIFICATIONS OF COSMETIC INGREDIENTS .......... 13 
3-2.1  Chemical identity ..................................................................................... 13 
3-2.2  Physical form ........................................................................................... 14 
3-2.3  Molecular weight ...................................................................................... 14 
3-2.4  Identification and purity of the chemical and isomer composition ................... 14 
3-2.5  Characterisation of the impurities or accompanying contaminants .................. 15 
3-2.6  Relevant physicochemical specifications ...................................................... 15 
3-2.7  Solubility ................................................................................................ 15 
3-2.8  Partition coefficient (Log Pow) ................................................................... 16 
3-2.9  Homogeneity and stability ......................................................................... 16 
3-3  EXPOSURE ASSESSMENT .............................................................................. 17 
3-3.1  Functions and uses of cosmetic ingredients ................................................. 17 
3-3.2  Identification of relevant exposure scenarios ............................................... 17 
3-3.3  Identification of the targeted dose for safety evaluation ................................ 17 
3-3.4  External exposure .................................................................................... 18 
3-3.4.1  Exposure models and tiered approach .................................................. 18 
3-3.4.1.1 Dermal exposure models ................................................................. 19 
3-3.4.1.2 Oral exposure models ...................................................................... 19 
3-3.4.1.3 Inhalation exposure models ............................................................. 20 
3-3.4.2  Model parameters .............................................................................. 23 
3-3.4.2.1 Daily use amounts and retention factors ............................................ 23 
3-3.4.2.2 Concentrations ............................................................................... 27 
3-3.4.2.3 Parameters specific for inhalation exposure ........................................ 27 
3-3.4.3  Aggregate exposure ........................................................................... 27 
3-3.5  Internal Exposure .................................................................................... 29 

 
 
 
 
 
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3-3.5.1  Toxicokinetics (ADME) ........................................................................ 29 
3-3.5.1.1 Dermal/percutaneous absorption ...................................................... 30 
3-3.5.1.2 Absorption after ingestion ................................................................ 33 
3-3.5.1.3 Inhalation ...................................................................................... 33 
3-3.5.2  Differences in metabolism for different routes ....................................... 34 
3-3.5.2.1 Systemic metabolism ...................................................................... 34 
3-3.5.2.2 Dermal metabolism ......................................................................... 35 
3-3.5.2.3 Lung metabolism ............................................................................ 35 
3-3.5.3  PBPK modelling ................................................................................. 36 
3-3.5.4  Calculation of the systemic exposure dose (SED) ................................... 38 
3-3.5.4.1 Calculation of the inhalation SED (SEDinh) ......................................... 39 
3-3.5.5  Aggregation of the systemic dose ........................................................ 40 
3-3.5.6  Human biomonitoring ......................................................................... 40 
3-3.5.6.1 Definition ....................................................................................... 41 
3-3.5.6.2 Fields of application ........................................................................ 41 
3-4  RELEVANT TOXICOLOGICAL TOOLS FOR THE SAFETY EVALUATION OF COSMETIC 
INGREDIENTS ...................................................................................................... 42 
3-4.1  New Approach Methodology (NAM) and Next-Generation Risk Assessment 
(NGRA) ............................................................................................................ 42 
3-4.2  Adverse Outcome Pathway (AOP)............................................................... 44 
3-4.3  In silico Assessment of Toxicological Hazard ................................................ 45 
3-4.3.1  In silico Toxicity Models ...................................................................... 45 
3-4.3.2  Read-across ...................................................................................... 46 
3-4.4  Acute toxicity .......................................................................................... 47 
3-4.4.1  Acute oral toxicity .............................................................................. 48 
3-4.4.2  Acute dermal toxicity ......................................................................... 48 
3-4.4.3  Acute inhalation toxicity ..................................................................... 49 
3-4.5  Skin corrosion and skin irritation ................................................................ 49 
3-4.5.1  Skin corrosion ................................................................................... 49 
3-4.5.2  Skin irritation .................................................................................... 49 
3-4.6  Serious eye damage and eye irritation ........................................................ 50 
3-4.7  Skin sensitisation ..................................................................................... 52 
3-4.7.1  Skin Sensitisation Quantitative risk assessment (QRA) ........................... 55 
3-4.7.2  Next-Generation Risk Assessment Approach (NGRA) .............................. 56 
3-4.8  Repeated dose toxicity.............................................................................. 57 
3-4.8.1  The use of uncertainty factors (UFs) for extrapolation for study duration .. 58 
3-4.8.2   Selection of PoD ................................................................................ 59 
3-4.9  Reproductive toxicity ................................................................................ 59 
3-4.10  Mutagenicity / Genotoxicity .................................................................... 60 
3-4.10.1 Definitions ........................................................................................ 60 
3-4.10.2 Mechanisms ...................................................................................... 61 
3-4.11  Carcinogenicity ..................................................................................... 68 

 
 
 
 
 
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3-4.12  Photo-induced toxicity ........................................................................... 70 
3-4.12.1 Photo-irritation and photo-sensitisation ................................................ 70 
3-4.12.2 Photomutagenicity / Photogenotoxicity ................................................. 71 
3-4.13  Human data in hazard assessment .......................................................... 73 
3-4.14  Other considerations ............................................................................. 73 
3-5  GENERAL PRINCIPLES FOR THE CALCULATION OF THE MARGIN OF SAFETY AND 
THRESHOLD OF TOXICOLOGICAL CONCERN ............................................................ 74 
3-5.1  Calculation of the Margin of Safety of a cosmetic ingredient .......................... 74 
3-5.1.1  The PoD value ................................................................................... 74 
3-5.1.1.1 Determination of NOAEL .................................................................. 74 
3-5.1.1.2 Determination of BMD ..................................................................... 75 
3-5.1.1.3 Choice of models ............................................................................ 75 
3-5.1.1.4 Adjustment factors to the PoD .......................................................... 76 
3-5.1.2  The PoDsys value............................................................................... 76 
3-5.1.3  MoS Analysis ..................................................................................... 76 
3-5.2  The threshold of toxicological concern (TTC) ................................................ 78 
3-5.2.1  General concept of TTC in risk assessment ............................................ 78 
3-5.2.2  TTC approach for human health risk assessment of chemical substances and 
cosmetic substances........................................................................................ 79 
3-5.2.3  iTTC APPROACH ................................................................................. 81 
3-6  SPECIAL CONSIDERATION FOR CERTAIN COSMETIC INGREDIENTS.................... 82 
3-6.1  Multi-constituent natural ingredients .......................................................... 82 
3-6.2  Identification of mineral, animal, botanical and biotechnological ingredients in a 
cosmetic product ............................................................................................... 83 
3-6.3  Animal-derived cosmetic substances .......................................................... 84 
3-6.4  Sun protection substances ........................................................................ 85 
3-6.5  Endocrine active substances (EAS) ............................................................. 85 
3-6.5.1  Definitions ........................................................................................ 85 
3-6.5.2  Identification of EDs and regulatory consequences ................................. 86 
3-6.5.3  Stepwise approach for cosmetics and their ingredients ........................... 86 
3-6.5.4  Cosmetic ingredients suspected to have ED properties ........................... 90 
3-6.6  CMR Substances ...................................................................................... 91 
3-6.7  Lifetime Cancer Risk (LCR) ........................................................................ 92 
3-6.8  Nanomaterials ......................................................................................... 92 
3-6.8.1  Definition of nanomaterial ................................................................... 92 
3-6.8.2  Potential safety issues of nanomaterials ................................................ 93 
3-6.8.3  Required information for nanomaterials ................................................ 95 
3-6.9  Hair dyes and hair dye components ............................................................ 96 
3-6.9.1  MoS calculations for hair dye formulations ............................................ 96 
3-6.9.2  Assessment of oxidative hair dye substances and reaction products ......... 96 
3-6.10  Cosmetic ingredients for baby and children products ................................. 97 
3-6.10.1 Definitions ........................................................................................ 97 

 
 
 
 
 
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3-6.10.2 Age-related susceptibilities/sensitivities ................................................ 98 
3-6.10.2.1 Dermal exposure of the newborn and early infant ............................ 98 
3-6.10.2.2 Cosmetic products used in the nappy area ..................................... 100 
3-6.11  Substances with very low dermal absorption ........................................... 100 
3-7   FURTHER REMARKS FOR APPLICANTS ............................................................ 100 
4.  REFERENCE LIST .............................................................................................. 102 
APPENDIX 1: INFORMATION ON REGULATION (EC) No 1223/2009 AND THE SCCS ......... 145 
APPENDIX 2: LISTS OF SUBSTANCES ....................................................................... 152 
APPENDIX 3: STANDARD FORMAT OF THE OPINIONS ................................................. 155 
APPENDIX 4: ANIMAL TESTING: INTERFACE BETWEEN REACH AND COSMETICS 
REGULATIONS ....................................................................................................... 164 
APPENDIX 5: CMR GUIDANCE ON SAFE USE OF CMR SUBSTANCES IN COSMETIC 
PRODUCTS ............................................................................................................ 165 
APPENDIX 6: REQUIREMENTS FOR THE CERTIFICATE OF ANALYSIS FOR A COSMETIC 
INGREDIENT .......................................................................................................... 169 
APPENDIX 7: DETAILED EXPOSURE DATA FOR COSMETIC PRODUCTS .......................... 170 
APPENDIX 8: KEY CHARACTERISTICS OF CARCINOGENS ............................................ 172 
APPENDIX 9: GUIDELINES ON MICROBIOLOGICAL QUALITY OF THE FINISHED COSMETIC 
PRODUCT .............................................................................................................. 174 
APPENDIX 10:  FREE ACCESS TO IN SILICO MUTAGENICITY/GENOTOXICITY AND 
CARCINOGENICITY DATABASES ............................................................................... 177 
APPENDIX 11: INHALATION PARAMETERIZATION ....................................................... 178 
APPENDIX 12: LIFETIME CANCER RISK (LCR) APPROACH ............................................ 179 
APPENDIX 13: PoD USED FOR TTC DERIVATION ........................................................ 180 
ABBREVIATIONS AND GLOSSARY OF TERMS ............................................................. 181 
 
 
 
   

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Nam et ipsa scientia potestas est

For knowledge itself is power

Francis Bacon (1561 - 1626) Essays

The “Notes of Guidance for the Testing of Cosmetic Ingredients and Their Safety

Evaluation by the SCCS” is a document compiled by the members of the Scientific

Committee on Consumer Safety (SCCS, replacing the former SCCP, SCCNFP and SCC).

The document contains relevant information on the different aspects of testing and

safety evaluation of cosmetic substances in Europe. The emphasis of this guidance is on

cosmetic ingredients, although some guidance is also indirectly given for the safety

assessment of finished products. It is designed to provide guidance to public authorities

and to the cosmetic industry in order to improve harmonised compliance with the

current cosmetic EU legislation. An important development in recent years was the full

implementation of the cosmetic legislation, Regulation (EC) No 1223/2009, meaning

that the animal testing and marketing bans fully apply from 2013 onwards: no in vivo

testing of finished products after 11 March 2004; no in vivo testing for local toxicity after

11 March 2009 and no in vivo testing for repeated dose toxicity (including sensitisation)

toxicokinetics and developmental toxicity from 11 March 2013 onwards for the purpose

of cosmetics. For this reason, the SCCS has closely followed the progress made with

regard to the development and validation of alternative methods, with emphasis on

replacement methodology.

The "Notes of Guidance" are regularly revised and updated in order to incorporate the

progress of scientific knowledge in general, and the experience gained, in particular in

the field of testing and safety evaluation of cosmetic ingredients.

The previous revision of the Notes of Guidance took place in 2018 (SCCS/1602/18).

Since then, several new addenda, opinions and memoranda of importance to the content

of this guidance document have been adopted and they form the basis of this new

revision. Focus is on exposure and the application of alternative methods, more

specifically on non-animal methods/new approach methodology (NAM).

As was also the case in previous revisions, individual opinions are not provided in detail

but, where relevant, are briefly summarised and clearly referred to.

The "Notes of Guidance" have been compiled to provide assistance in the complex

process of the testing and safety evaluation of cosmetic ingredients in the EU.

Input of scientists from the Scientific Committee on Health and Environmental and

Emerging Risks (SCHEER) and Cosmetics Europe (CoE) is gratefully acknowledged.

The Chairperson

ACKNOWLEDGMENTS

SCCS members listed below are acknowledged for their valuable contribution to the

finalisation of this guidance document.

SCCS members

Dr U. Bernauer

Dr L. Bodin

Prof. Q. Chaudhry (SCCS Chair)

Prof. P.J. Coenraads (SCCS Vice-Chair)

Prof. M. Dusinska

Dr J. Ezendam

Dr E. Gaffet

Prof. C. L. Galli

Dr B. Granum

Prof. E. Panteri

Prof. V. Rogiers (SCCS Vice-Chair and rapporteur)

Dr Ch. Rousselle

Dr M. Stepnik

Prof. T. Vanhaecke

Dr S. Wijnhoven

SCCS external experts

Dr A. Koutsodimou

Prof. W. Uter

Dr N. von Goetz

All Declarations of Working Group members are available on the following webpage:

The SCCS Notes of Guidance document is not open for commenting period as it

remains a living document, which is regularly updated. Any observation may be

sent to SCCS mailbox ([email protected]) for further consideration by

the SCCS.

Keywords: SCCS, SCCS Notes of Guidance for the Testing of Cosmetic Ingredients and their

Safety Evaluation, 11

revision, SCCS/1628/21.

Opinion to be cited as: SCCS (Scientific Committee on Consumer Safety), SCCS Notes of

Guidance for the Testing of Cosmetic Ingredients and their Safety Evaluation 11

revision,

30-31 March 2021, SCCS/1628/21.

_____________________________________________________________________________________________

About the Scientific Committees

Two independent non-food Scientific Committees provide the Commission with the scientific

advice it needs when preparing policy and proposals relating to consumer safety, public health

and the environment. The Committees also draw the Commission's attention to the new or

emerging problems which may pose an actual or potential threat.

They are the Scientific Committee on Consumer Safety (SCCS) and the Scientific Committee

on Health, Environmental and Emerging Risks (SCHEER), and they are made up of

independent experts.

In addition, the Commission relies upon the work of the European Food Safety Authority

(EFSA), the European Medicines Agency (EMA), the European Centre for Disease prevention

and Control (ECDC) and the European Chemicals Agency (ECHA).

SCCS

The Committee shall provide Opinions on questions concerning health and safety risks

(notably chemical, biological, mechanical and other physical risks) of non-food consumer

products (for example cosmetic products and their ingredients, toys, textiles, clothing,

personal care and household products such as detergents, etc.) and services (for example:

tattooing, artificial sun tanning, etc.).

Scientific Committee members

Ulrike Bernauer, Laurent Bodin, Qasim Chaudhry, Pieter Jan Coenraads, Maria Dusinska,

Janine Ezendam, Eric Gaffet, Corrado Lodovico Galli, Berit Granum, Eirini Panteri, Vera

Rogiers, Christophe Rousselle, Maciej Stepnik, Tamara Vanhaecke, Susan Wijnhoven

Contact:

European Commission

Health and Food Safety

Directorate C: Public Health

Unit C2: Health information and integration in all policies

L-2920 Luxembourg

[email protected]

European Union, 2022

PDF ISSN 1831-4767 ISBN 978-92-76-54826-3 doi:10.2875/273162 EW-AQ-22-017-EN-N

The opinions of the Scientific Committees present the views of the independent scientists who

are members of the committees. They do not necessarily reflect the views of the European

Commission. The opinions are published by the European Commission in their original

language only.

https://health.ec.europa.eu/scientific-committees/scientific-committee-consumer-safety-

sccs_en

_____________________________________________________________________________________________

Applicants are invited to visit the SCCS website:

https://ec.europa.eu/health/scientific_committees/consumer_safety/opinions_en

where Applicants will find a checklist

for submitting a safety dossier of a cosmetic ingredient.

Applicants are invited to visit the following website for further legislative information:

https://ec.europa.eu/growth/sectors/cosmetics/legislation_en

_____________________________________________________________________________________________

MAIN CHANGES IN 11

REVISION OF THE SCCS NOTES OF GUIDANCE (NOG)

The whole NoG have been revised and updated with a particular emphasis on the following:

- Inhalation models

- In silico methodology for genotoxicity/carcinogenicity

- General updating of NAMs

- Scientific concerns for the safety of the nanomaterials

- Update of ED section

- Discussion on uncertainty factors

- Updating of references in general

- Appendix 1: complying with the testing & marketing bans

- Appendix 9: guidelines on microbiological quality of the finished product

- Appendix 10: free access to in silico mutagenicity/genotoxicity databases

- Appendix 11: inhalation parameterisation

- Appendix 12: Lifetime Cancer Risk Approach

- Appendix 13: PoD used for TTC derivation

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1. INTRODUCTION

Since July 2013, Regulation (EC) No 1223/2009 applies for cosmetic products. Their safety-

in-use is, as was also the case for Directive 76/768/EEC, established by controlling the safety

of the ingredients.

For those ingredients for which some concern exists with respect to human health (e.g.

colourants, preservatives, UV-filters, hair dyes), safety evaluation is done at the Commission

level by the Scientific Committee on Consumer Safety (SCCS). These substances are

addressed in the Annexes of Regulation (EC) No 1223/2009.

For the safety evaluation of cosmetic ingredients, all available scientific data are considered,

taking into account the testing and marketing bans in force under Regulation (EC) No

1223/2009. This includes the physical and chemical properties of the compounds under

investigation, exposure via relevant exposure routes, in silico data such as results obtained

from (Q)SAR {(Quantitative) Structure Activity Relationship} modelling, chemical categories,

grouping, read-across, Physiologically Based PharmacoKinetics (PBPK) /ToxicoKinetics (PBTK)

modelling, in vitro and ex vivo experimental results and data obtained from animal studies

(in vivo) that have been carried out for the purpose of cosmetics before the testing and

marketing bans. The animal testing ban on finished cosmetic products applies since 11

September 2004; the testing ban on ingredients or combination of ingredients applies since

11 March 2009. The marketing ban applies since 11 March 2009 for all human health effects

with the exception of repeated-dose toxicity, reproductive toxicity, and toxicokinetics. For

these specific health effects, the marketing ban applies since 11 March 2013, irrespective of

the availability of alternative non-animal methods. In addition, clinical data, epidemiological

studies, information derived from accidents, data from Post-Marketing Surveillance (PMS) or

other human data are also taken into consideration

In the present update, the state-of-the-art with respect to the validated methods of the 3Rs

(Refinement, Reduction and Replacement) strategy of Russell et al. (1959), is incorporated

with emphasis on New Approach Methodologies (NAMs). In view of the testing and marketing

bans in the cosmetic regulation, the SCCS gives special attention to those alternative methods

that are suitable for the safety testing of cosmetic substances. New methodologies for risk

assessment of chemicals without using animal experimentation are worldwide being explored.

Attention is given here to Next-Generation Risk Assessment (NGRA) as a possible framework

for the safety evaluation of cosmetic ingredients and the NAMs that would fit into this

structure (Rogiers et al., 2020). Risk assessment of cosmetics and their ingredients is shifting

towards a strategic combination of NAMs and new technology with historical animal data, if

available, to come to a Weight of Evidence (WoE) decision making approach.

Although the "Notes of Guidance" are concerned with the testing and safety evaluation of the

cosmetic substances listed in the Annexes of Regulation (EC) No 1223/2009 and those for

which safety concerns have been expressed, they could be also of interest for all substances

intended to be incorporated in a cosmetic product. Even though the "Notes of Guidance" have

not been written for the latter purpose, they can indeed be of practical use in making a

Product Information File (PIF) for a finished cosmetic product as currently required by

Regulation (EC) No 1223/2009.

The European Chemicals Agency (ECHA) can ask for animal studies even if the substance is

foreseen only for cosmetic use (see Appendix 1, section 3). The applicant can submit these

animal data to ECHA, but cannot use these in the cosmetic product safety report (CPSR) for

the product information file (PIF) and cannot submit these to the SCCS for risk assessment

of the ingredient under cosideration. If SCCS knows about the existence of such a file at

ECHA, they can request access to these studies and consider whether the results have an

impact on the risk assessment of the substance and change their view.

The “Notes of Guidance” should not be seen as a prescriptive procedure, but rather as an

approach that may need to be adapted on a case-by-case basis when evaluating the safety

_____________________________________________________________________________________________

of the Annex substances. However, when major deviations from standardised

protocols/procedures in the safety evaluation process have been adopted, it is essential that

Applicants provide scientific justification.

The "Notes of Guidance" will be revised as scientifically required on the basis of scientific

advances in toxicology and validated alternative methods or legislative changes.

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2. THE SCIENTIFIC COMMITTEE ON CONSUMER SAFETY, SCCS

2-1 BACKGROUND

The Commission Decision C(2015)5383 of 7.8.2015

established the new Scientific

Committees in the field of public health, consumer safety and the environment. Members

were appointed

for a five-year term (2016-2021) and a reserve list

was created. The term

was extended until end of 2026 due to Covid-19. The Principles and Working Procedures of

the Scientific Committees are stated in their establishing Decision and in the Rules of

Procedure adopted by their members (April, 2016)

For more information, see Appendix 1.

2-2 MANDATE

The SCCS is an advisory body that provides the Commission with scientific advice and safety

evaluations for Annex substances and compounds for which some concern for human health

exists. Its consultation for this task is compulsory.

For more information, see Appendix 1.

2-3 RULES OF PROCEDURE

The SCCS works with 3 working groups, dealing with:

− cosmetic ingredients

− methodology

− nanomaterials.

Safety evaluations and advice are taken up in opinions, which are adopted during a plenary

meeting (or by written procedure). A commenting period of minimum four weeks (later

agreed on eight weeks) is foreseen for draft opinions before they are finalised and published.

For more information, see Appendix 1.

2-4 OPINIONS

Opinions are published on the SCCS website:

https://ec.europa.eu/health/scientific_committees/consumer_safety/opinions_en.

For more information, see Appendix 1.

2-4.1 THE "NOTES OF GUIDANCE"

One of the responsibilities of the SCCS is to recommend a set of guidelines to be taken into

consideration by the cosmetic and raw material industry in developing adequate studies to

be used in the safety evaluation of cosmetic substances.

This is done through the ‘Notes of Guidance for the Testing of Cosmetic Ingredients and Their

Safety Evaluation’ (NoG) that are regularly revised and updated in order to incorporate new

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/call_2015_5383_decision_with_annexes

_en.pdf

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/appointment_letter_2016_en.pdf

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/appointment_reserve_list_2016_en.pdf

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/rules_procedure_2016_en.pdf

_____________________________________________________________________________________________

knowledge and scientific and regulatory advances. Therefore, dossiers submitted to the SCCS

should be in accordance with the latest published version of the NoG. The 10

Revision

SCCS/1602/18 is now replaced by this 11

Revision SCCS/1628/21.

As cosmetic ingredients are chemical substances, the NoG include the toxicological test

procedures reported in Commission Regulation (EC) No 440/2008. The latter describes the

basic toxicity testing procedures needed to evaluate different human health-related

toxicological endpoints and are internationally accepted as being the result of long-term

scientific agreement. Whereas the testing procedures for chemical substances take the 3Rs-

principle into consideration, animal experiments for cosmetic purposes are excluded in the

EU. For the safety evaluation of cosmetic ingredients only validated non-animal

methods/NAMs may be applied. Furthermore, testing procedures in accordance with the

Organisation for Economic Co-operation and Development (OECD) Guidelines, and, on a case-

by-case basis, well documented scientifically justified alternative methods that may not have

been officially validated yet are also carefully considered. Data obtained from animal

experimentation for the purpose of cosmetics or other consumer products legislation and

generated before the established cosmetic deadlines of the testing and marketing bans (see

1. Introduction) still may be used in the safety evaluation of cosmetics and their ingredients.

As regards data generated after the deadlines of the testing and marketing bans, see Section

3 of Appendix 1.

For the SCCS’ safety evaluation, the systemic doses obtained (mostly) after oral

administration are used. For local toxicity endpoints, normally only hazard identification is

carried out. Safety evaluation is done for intact skin.

2-4.2 SCCS COSMETIC INGREDIENT DOSSIERS

Regulation (EC) No 1223/2009 requires Annexed cosmetic substances to be notified, safety

assessed and adequately labelled before being allowed on the EU market. These annexes lay

down clear limitations and requirements for the cosmetic substances concerned. The safety

assessment of the cosmetic ingredients in the EU is overseen by the SCCS. The evaluations

carried out by the SCCS are based on safety dossiers submitted by Applicants (individual

company/associations, Competent Authorities).

In view of the animal testing and marketing bans of cosmetic ingredients/products, two main

routes to developing safety dossiers are possible:

● In case a new ingredient is to be used exclusively in a cosmetic product, testing needs

to be in compliance with the restrictions on animal testing placed under Regulation

(EC) No 1223/2009 and safety data need to be derived from non-animal alternative

methods/NAMs.

● When an ingredient has pre-existing safety data derived from animal tests (e.g. an

existing cosmetic ingredient) that have been carried out before the regulatory

deadlines, it can still be used.

● Animal test data relating to chemical substances to be used also in products other

than cosmetics (e.g. food, medicines, biocides, etc.) can also be used for supporting

safety assessment of an ingredient intended to be used in a cosmetic product.

Further information is provided in Section 3 of Appendix 1.

In case of a negative or inconclusive opinion by the SCCS, resubmission of a dossier

is only possible when the Applicant provides sufficient (new) evidence to address

the concerns raised.

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2-4.3 SPECIFIC ISSUES TAKEN UP IN NOG

In addition to the regular revision of the NoG and the study of toxicological dossiers of

cosmetic substances for inclusion in one of the Annexes of Regulation (EC) No 1223/2009,

in the following sections some specific issues are addressed. Examples include (non-

exhaustive list):

• New Approach Methodology (NAM) in the safety assessment of cosmetic ingredients

• Introduction to Next Generation Risk Assessment (NGRA)

• Threshold of Toxicological Concern (TTC)

• Endocrine disruptors’ issues

• CMR (Carcinogenic, Mutagenic, toxic to Reproduction) issues

• Safety assessment of hair dyes and colourants

• Safety assessment of nanomaterials

• Safety of cosmetic ingredients for babies and children

• Fragrance allergy in consumers

• Risk and health effects: miscellaneous

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3. SAFETY EVALUATION OF COSMETIC INGREDIENTS

3-1 SAFETY EVALUATION OF COSMETIC INGREDIENTS AS APPLIED BY THE SCCS

- The safety of cosmetic products is based on the safety of the ingredients

The rationale behind the safety of the cosmetic product being based on the safety of its

ingredients comes from the fact that many thousands of different cosmetic products on the

EU market are all derived from a limited number of substances. Hence, toxicity testing has

been concentrated on ingredients, and particularly on those that are intended to react with

biological moieties and therefore are of potential concern for human health. This is also the

basis for the lists of authorized, banned and restricted substances (Table 1).

Annex II

List of prohibited substances

Annex III

List of restricted substances

Annex IV

List of allowed colourants

Annex V

List of allowed preservatives

Annex VI

List of allowed UV-filters

Table 1: Annexes to Regulation (EC) No 1223/2009

- For the safety evaluation of cosmetic ingredients two channels are functional

The safety of the Annex substances is evaluated by the SCCS; the safety of cosmetic

products with all their ingredients is evaluated by the industry placing them on the EU

market. Thus, the Annex substances fall under the responsibility of the SCCS (left part of

Figure 1). All the ingredients in cosmetic products are the responsibility of the “Responsible

Person, RP”, as defined by Regulation (EC) No 1223/2009, through the safety assessor (right

part of Figure 1).

Figure 1: Human health safety evaluation of cosmetic ingredients in the EU.

PIF: Product Information File

_____________________________________________________________________________________________

- This guidance, in principle, equally applies to the safety evaluations carried out

by the SCCS as by the safety assessors of the cosmetic industry.

Safety evaluation is generally performed taking into account the data provided by the industry

or in some cases by Members States authorities. The SCCS also has the opportunity to add

relevant data from the open literature or other relevant sources.

In general, the safety evaluation of cosmetic ingredients by the SCCS is based upon the

principles and practice of the risk assessment process universally applied for chemical

substances with the challenge that only validated replacement methods (or demonstrated to

be scientifically valid) should be used when testing for the purposes of the EU cosmetic

legislation.

A typical safety evaluation procedure comprises the following elements:

1) Hazard identification is carried out to identify the intrinsic toxicological properties of the

substance, i.e. whether it has the potential to damage human health. It is based on the

results of in vivo studies, in vitro and ex vivo tests, in chemico methodology, in silico

methods and read-across, clinical studies, case reports, epidemiological studies and data

from Post-Marketing Surveillance (PMS). Intrinsic physical and chemical properties of the

substance under consideration are also taken into account.

2) Exposure assessment

Human exposure is calculated based on the declared functions and uses of a substance

as a cosmetic ingredient, the amount present in the respective cosmetic product

categories and their frequency of use.

The single product exposure describes the exposure to a cosmetic ingredient in one

product category via one route.

The aggregate exposure, in the context of the NoG, is the sum of all relevant single

product exposures, so that it describes the exposure from all product categories in which

the cosmetic ingredient is used and all relevant exposure routes.

Where necessary, exposure of vulnerable consumer groups could be assessed separately

(e.g. children, pregnant woman, etc.).

Generally, only exposures from the use of a substance as cosmetic ingredient are

considered, with the exception of CMR compounds, for which non-cosmetic uses should

also be taken into account (see section 3-6.6 and Appendix 5).

3) Dose-response assessment

For the relationship between the exposure and the toxic response, a Point of Departure

(PoD) is determined. The PoD is defined as the dose-response point that marks the

beginning of a low-dose extrapolation (for threshold and non-threshold compounds). In

most Opinions a No Observed Adverse Effect Level (NOAEL) has been used as PoD.

The SCCS considers that, where usable in vivo data are available, the preferred method

for both threshold and non-threshold cosmetic ingredients is to express the dose metric

as BenchMark Dose (BMD). Both the European Food Safety Agency (EFSA) and the World

Health Organization (WHO) also recommend that the BMD approach for deriving the PoD

should be used as a starting point for human health risk assessment.

The BMD approach has a number of advantages over using NOAEL. It makes complete use of

the available dose - response data

- it takes into account the shape of the dose - response curve

- it is less dependent on dose spacing

_____________________________________________________________________________________________

- it enables quantification of the uncertainties in the dose - response data using

statistical methodology (EFSA, 2016).

For compounds with a threshold, the PoD can be a NOAEL, a Lowest Observed Adverse

Effect Level (LOAEL), or a BMD Lower limit (BMDL) (for details of the NOAEL and BMD

approach, see Sections 3-4.8, 3-5.1)

4) Risk characterisation

In risk characterisation, the focus in the NoG is on systemic effects. In the case of a

threshold effect, the Margin of Safety (MoS) is mostly calculated from oral toxicity

studies, unless robust dermal toxicity data are available

. In the case of an oral toxicity

study, the following equation (1) is used:

PoD

sys

MoS = (1)

SED

The PoD

sys

is a dose descriptor for the systemic exposure to a substance and is calculated

from the oral PoD by use of the proportion of the substance systemically absorbed. SED

represents the Systemic Exposure Dose (see also Section 3-3.5.4). In this equation, PoD

BMDL or, alternatively, NOAEL or LOAEL, where BMDL cannot be calculated.

For non-threshold effects (e.g. a non-threshold carcinogenic effect), the lifetime risk is often

based on the BMD10 (benchmark dose response for a 10% response). The risk assessment

of carcinogens is described in Section 3-4.11.

Risk characterisation is followed by risk management and risk communication, which are not

in the remit of the SCCS, but of the European Commission or the RP, the latter when a finished

cosmetic product and its ingredients are involved (Figure 1).

Besides the normal procedure when the industry or Member States or their representatives

submit a complete dossier, in some cases, either upon request of the Commission or on a

voluntary basis, industry provides additional data on cosmetic ingredients that have been

assessed in the past. An evaluation exclusively based on additional reports, together with

summaries of earlier submissions, however, may not be adequate. Therefore, complete

dossiers may be required case by case, even though a re-evaluation of only a part of a dossier

appears necessary. Dossiers and full studies should be submitted in common formats such

as pdf or Word and need to be readable and searchable.

Other common formats that allow copy/paste actions are accepted. Scanned documents that

are not readable/ searchable are not accepted.

It is beyond the scope of the NoG to discuss the whole process of risk assessment. Numerous

review articles and textbooks exist on this topic. The aim is to highlight some key aspects to

explain why certain data and test results should be provided in the dossiers on the cosmetic

substances presented to the SCCS for evaluation.

An example of the framework of a typical dossier is given in Appendix 3.

For the case that a dermal repeated dose toxicity study is used, see Section 3-4.8 and 3-5.1

_____________________________________________________________________________________________

The contact point for dossier submissions and regulatory/risk management questions is:

GROW-F2@ec.europa.eu

The SCCS address for scientific requests is: SANTE-C2-SC[email protected]pa.eu

3-2 CHEMICAL AND PHYSICAL SPECIFICATIONS OF COSMETIC INGREDIENTS

Physical and chemical properties of substances are considered as crucial information, since

they may indicate potential risks. For example, a small Molecular Weight (MW) hydrophobic

compound is more likely to penetrate through the skin than a high MW hydrophilic compound.

Physical and chemical properties also identify physical hazards of the substance (e.g.

corrosiveness as indicated by pH of aqueous solution, volatility, explosiveness, flammability).

In addition, some QSAR (Quantitative Structure Activity Relationship) programmes and

empirical models require physical and chemical property values as inputs for in silico

estimation of properties and potential biological effects.

The basic and minimal specifications for any cosmetic ingredient to be evaluated are:

1) Chemical identity;

2) Physical form;

3) MW;

4) Characterisation and purity of the chemical, including isomer composition whenever

relevant for safety assessment;

5) Characterisation of the impurities or accompanying contaminants;

6) Solubility;

7) Partition coefficient (Log P

);

8) Vapour pressure (volatile liquids);

9) Homogeneity and stability;

10) Further physical and chemical properties if relevant for safety evaluation.

For nanomaterials, special requirements for provision of physicochemical data apply (see

Section 3-6.8). Original data on all these points must be included in each toxicological dossier

and information and documentation for all analytical data should be provided.

The appropriate certificate of analysis must also be presented for the test chemical used to

generate the data as submitted in the dossier to the SCCS.

Preference is clearly given to measured parameters of relevant batches on the market over

calculated values (e.g. log P

) or literature data (where often batches are tested that differ

from the batches used in toxicological tests and therefore may have different composition /

impurity profiles).

In the following section, the methods are (where relevant) accompanied by their

corresponding reference number in Regulation (EC) No 440/2008 (2008/440/EC).

3-2.1 CHEMICAL IDENTITY

The precise identity and chemical nature of the substance under consideration and its

structural formula must be given. The Chemical Abstracts Service (CAS) number of the

chemical, the International Nomenclature of Cosmetic Ingredients (INCI) name or Common

Ingredient Nomenclature (CIN) name and the EC number (see Appendix 2 for more details)

should be provided.

With regard to substances that cannot be identified in terms of their structural formula,

sufficient information should be provided on the method of preparation (including all physical,

_____________________________________________________________________________________________

chemical, enzymatic, (bio)technological or microbiological steps) and the materials used in

their preparation to enable assessment of the probable structure and activity of the

compound(s).

For the safety evaluation of a complex mixture (e.g. an extract), complete information should

be provided on the origin of the source materials (e.g. part of a plant), extraction method

and any additional processes and/or purification steps used (see Section 3-6.1) to establish

a standardised material as representative of the extract present in commercial products.

In case of a mixture, components must be described in terms of qualitative and quantitative

formulae. These could be: main components, preservatives, antioxidants, chelators, buffering

agents, solvents, other additives, impurities and/or additional external contamination.

When a cosmetic ingredient and its derivatives (salt, ester, …) are submitted for evaluation,

this must be clearly specified in the dossier, because the chemical form can determine the

safety evaluation. The physical and chemical properties of all specific chemical forms must be

provided, and the same specific substances must be used in the toxicological studies

performed for the safety evaluation. Any deviations must be justified.

3-2.2 PHYSICAL FORM

A description of the physical form should be given: powder, paste, gel, liquid. For

nanoparticles, further information as specified in Section 3-6.8 should be given, including the

particle size and its distribution.

For polymer ingredients, the molecular weight distribution should be provided.

3-2.3 MOLECULAR WEIGHT

The MW of each substance should be given in Daltons. In the case of mixtures, the MW must

be given for the constituents.

3-2.4 IDENTIFICATION AND PURITY OF THE CHEMICAL AND ISOMER COMPOSITION

The degree of purity must be clearly indicated. The validity of the analytical methodology

used must be shown. When a reference material/standard is used for the determination of

purity, a certificate of analysis of the reference standard should be submitted (Appendix 6)

Purity of the active substance based on High Performance Liquid Chromatography (HPLC)

peak area can only be accepted when:

1) a reference material of known purity is used,

2) the HPLC recovery of the test material is clearly documented,

3) the ultraviolet (UV) detection of the active substance is performed at λ

max

, in an

appropriate mobile phase, and

4) peak purity of the active substance is clearly documented.

The experimental conditions of the techniques used for the chemical characterisation UV,

Infra Red (IR) and Nuclear Magnetic Resonance (NMR) spectroscopy, Mass Spectrometry

(MS), chromatographic techniques e.g. Gass Chromatography (GC), elemental analysis, etc.)

as well as the resulting spectra, chromatograms etc. should be provided.

The substance(s) used in physical and chemical tests, toxicity studies, etc., mentioned in the

dossier, must be either exactly the same material(s) under consideration or justifiably

representative of the substances present in commercial products.

_____________________________________________________________________________________________

When a substance is a mixture of isomers, only the relevant isomer(s) used as a cosmetic

ingredient should be included in the safety assessment. The other isomer(s) is/are considered

as an impurity or impurities. Information on isomer composition should be provided.

3-2.5 CHARACTERISATION OF THE IMPURITIES OR ACCOMPANYING CONTAMINANTS

In addition to the purity of the substance under consideration, identity in terms of the

chemical nature and concentration of impurities that may be present must also be stated.

Impurities should be characterised and quantified by an appropriate analytical method, e.g.

by HPLC-PDA (Photometric Diode Array), LC-MS/GC-MS, NMR spectroscopy etc., using

reference standards with documented purity, where appropriate. Validated analytical

procedures should be used for impurity testing. There is no specific recommendation available

to assess the limit of acceptable non-CMRs impurities for cosmetic products.

Small changes in the nature of some impurities may considerably alter the toxicity of

substances. In general, results of safety studies on a particular substance are only relevant

when they refer to that substance used, with its own specific purity and impurity profile. The

scientific validity of tests performed on batches of the substance with diverging purities

deserves careful interpretation. Therefore, it must be ensured that neither other impurities

nor an increased level of impurities are present in the representative commercial material.

For this, the stability of the synthesis process, including any purification measures, is

important. A change in these processes will need careful re-evaluation of the impurities, even

if the level of purities remains the same.

3-2.6 RELEVANT PHYSICOCHEMICAL SPECIFICATIONS

A typical physicochemical dataset consists of:

- Physical state (solid, liquid, gas)

- Organoleptic properties (colour, odour, taste if relevant)

- Solubility (EC A.6) in water and relevant solvents, including receptor fluids (at … °C)

- Partition coefficient (EC A.8) (Log P

, at … °C), if applicable

- Flash point (EC A.9)

- Physical properties depending on the physical state:

o for liquids: boiling point (EC A.2), relative density (EC A.3) (at … °C), pK

(at … °C),

viscosity (at … °C), vapour pressure [EC A.4] (at … °C), ....

o for solids: morphological form (crystal form, amorphous, ...), melting temperature

(EC A.1), pK

(…% in ..., at … °C), ...

o for gases: density (EC A.3) (at … °C and pressure), auto-ignition temperature (EC

A.15)

- In case of a UV-absorbing substance, the UV-absorption spectrum of the compound should

be included. It is self-evident that for UV filters, the UV spectrum is indispensable.

- For nanomaterials and nanoparticles special requirements apply (see Section 3-6.8).

3-2.7 SOLUBILITY

The solubility (EC A.6) of the substance in water and/or in any other relevant organic solvent

should be stated (in g/l at … °C). Some substances are sparingly soluble or insoluble in

aqueous media or other solvents. These should be clearly stated. In Table 2, different

solubility terms have been defined.

_____________________________________________________________________________________________

Where the solubility of the active substance in water is low (according to EU Method A.6), a

highly sensitive and selective analytical technique (such as LC/MS) should also be used to

document the solubility and to rule out that the soluble material may be an impurity (or

impurities) in the test material. Similarly, solubility of substances that are poorly soluble in

various solvents should be measured by highly sensitive and selective analytical technique

(such as LC/MS). In cases of low solubility of the active substance in reverse phase HPLC

mobile phases, sensitive detection systems, such as MS, should be applied, or another normal

phase chromatography should be used.

The solubility of the active substance in the solvent systems used in various studies should

also be clearly presented.

Table 2: Definition of solubility terms (adapted from USP38/ USP38–NF33*and General

Notices (Ph. Eur. 10

Ed.)

Term*

Parts of Solvent Required

for 1 Part of Solute*

Solubility defined in g/L

(deduced by SCCS)

Very soluble

Less than 1 part

>1000

Freely soluble

1 to 10 parts

100-1000

Soluble

10 to 30 parts

33.3-100

Sparingly soluble

30 to 100 parts

10-33.3

Slightly soluble

100 to 1000 parts

1-10

Very slightly soluble

1000 to 10000 parts

0.1-1

practically insoluble, or

insoluble

>10000, or equal to10 000

parts

< 0.1 or = 0.1

*Under USP38/ USP38–NF33: practically insoluble is used in USA; in EU: insoluble

3-2.8 PARTITION COEFFICIENT (LOG POW)

The n-octanol/ water partition coefficient (EC A.8) should be given, along with the pH and

temperature conditions.

In the case of a calculated value, the method used for estimation should be specified.

LogP

values often depend on the pH, especially for ionisable molecules, zwitterions, etc.

Therefore, a single calculated value of Log P

, without any reference to the respective pH,

cannot be correlated to the physiological conditions and the pH conditions of the dermal

absorption studies.

3-2.9 HOMOGENEITY AND STABILITY

Homogeneity data of the test solutions with respect to the content of the test substance,

under experimental conditions, should be provided.

Data on the stability of the test substance under the experimental conditions of the reported

studies and under conditions of use should be provided. Validated analytical procedures

should be used to determine stability of the test substance. In addition, the stability of the

test substance relating to its thermal stability and, if applicable, sensitivity to moisture or

oxygen under storage conditions and in typical cosmetic formulations should also be provided.

Any degradation products should be chemically characterised. In this regard, it is important

that the storage conditions and the lengths of studies chosen should be sufficient to cover

the storage, shipment, and subsequent use. The stability studies should also be conducted

on the test substance packaged in a container, which is the same as the container intended

for storage and distribution for marketing.

_____________________________________________________________________________________________

3-3 EXPOSURE ASSESSMENT

3-3.1 FUNCTIONS AND USES OF COSMETIC INGREDIENTS

For substances that are evaluated as cosmetic ingredients, the concentration, function and

way of achieving that function in marketed cosmetic products should be reported. In

particular, it should be explicitly mentioned whether substances are meant to be included in

sprays or aerosols since consumer exposure via inhalation is then probable and needs to be

taken into consideration in the overall risk assessment.

In addition, other uses of the substance (e.g. in consumer products, industrial products) and,

wherever possible, the concentrations involved in such uses should be described.

3-3.2 IDENTIFICATION OF RELEVANT EXPOSURE SCENARIOS

In order to assess exposure of the end users, relevant exposure scenarios have to be

identified that comprise all the important functions and uses of a cosmetic ingredient (see

Section 3-3.1). These scenarios need to describe "reasonably foreseeable exposure

conditions" (Cosmetics Regulation (EC) No 1223/2009, Article 16 f), under which these the

cosmetic product should be safe.

The following parameters describe an exposure scenario. However, the list is not exhaustive,

and further parameters may need to be taken into account. Note that all routes of exposure

(dermal, oral and inhalation) should be considered in view of the intended use of the product.

− cosmetic product type (s) in which the ingredient may be used

− method of application as detailed as possible, e.g. rubbed-on, sprayed, applied and

washed off, etc.; considerations whether the product is a rinse-off or leave-on product

and which retention factor should be applied

− concentration of the ingredient in the marketed cosmetic product

− quantity of the product used at each application

− frequency of use

− total area of skin contact

− duration of exposure

− target consumer groups (e.g. children, people with sensitive, damaged or

compromised skin) where specifically required

− application on skin areas exposed to sunlight

− location of use (indoors/outdoors) and ventilation

3-3.3 IDENTIFICATION OF THE TARGETED DOSE FOR SAFETY EVALUATION

The hazard identification can either point to systemic effects that require comparison to a

SED or local effects, like skin/eye irritation, skin sensitisation, sun-induced skin reactions or

effects on the lungs, which mostly are dependent on the amount of substance acting on the

surface tissues of the respective body part and require comparison to a Local External Dose

(LED).

In the exposure assessment, first the LEDs are calculated that are expected at the specific

body entrances and available for uptake. The most important body entrances for substances

in cosmetics are the skin, the inhalatory tract and the mouth. These correspond to the uptake

routes for internal exposure (dermal route, inhalation route and oral ingestion). For selected

_____________________________________________________________________________________________

products other entrances are possible, e.g. via the eyes (e.g. eye makeup), or via genital

regions (e.g. intimate spray, intimate creams).

As an example, the LED in the lung (the amount of compound per g of lung tissue) can be

compared to a “local” NOAEL, and a local MoS can be calculated for effects on the lungs.

The external exposure can further be used to calculate internal (or systemic) exposure which

corresponds to an internal dose (see Section 3-3.5.4). For the calculation of the SED,

absorption (or uptake) specific to the respective exposure route has to be taken into account.

For risk assessment, the MoS (see Section 3-5.1) is based on the internal dose, i.e. the SED.

3-3.4 EXTERNAL EXPOSURE

3-3.4.1 EXPOSURE MODELS AND TIERED APPROACH

Exposure is calculated based on exposure scenarios by using appropriate exposure models.

Generally, external exposure is calculated by multiplying the concentration/fraction of a

substance in a source with the amount of the source that is applied on, or reaches, a specified

site. To save time and resources, a tiered approach is normally followed that first

investigates exposure based on generic exposure scenarios with conservative point values as

model parameters (screening level).

Where necessary, these conservative exposure estimates are refined in a higher tier by using

probabilistic approaches or other means of refinement (Meek et al., 2011).

For the safety evaluation of cosmetics, such a screening level approach is the calculation of

aggregate exposure according to the NoG. The parameter values presented there can be used

as the basis for a deterministic first-tier assessment. If a refinement is necessary, a

probabilistic approach can be followed by the use of appropriate models and/or tools.

However, this needs to be clearly justified. For regulatory purposes, the probabilistic approach

needs to be conservative but realistic and transparent.

In particular, for probabilistic assessments the SCCS recommends the following:

- Habits and practices in a population regarding the use of product categories may be

treated probabilistically, under the assumption that they will not change rapidly over time.

- The target protection goal will be the 95

percentile of the European population.

Therefore, for a probabilistic assessment the relevant SED for deriving the MoS will be the

percentile of the probabilistically assessed population exposure.

- Ingredient concentrations in product categories should normally cover the worst case, i.e.

for ingredients with restrictions on concentrations and applicability domains (Annex III of

the EU Cosmetic Regulation), also in the probabilistic assessment the maximal allowed

concentrations should be used, and for other ingredients the maximal concentrations that

are realistically foreseeable in a specific product category. This is because product

formulations may be highly variable over time, so that an assessment of ingredient

concentrations at a specific point in time may not cover the use of the ingredient in the

future.

- For reasons of transparency, the model equations and the input parameters need to be

provided together with the exposure estimates, so that the exposure calculation is

reproducible. If this is not possible, because a specific tool has been used, the original

input file containing used distributions and all settings, and the original output file need

to be provided by the Applicant. The output file needs to contain the date of the

assessment, the relevant model settings and parameters for this assessment and the

associated results, ideally not only in tabular form by giving relevant percentiles of the

exposure distribution, but also by graphical visualisation.

_____________________________________________________________________________________________

3-3.4.1.1 DERMAL EXPOSURE MODELS

For cosmetics, the dermal route is often the most important one.

Apart from the general approach, the calculation of dermal exposure needs to take into

account that only a fraction of the product is retained on the skin. Therefore, a retention

factor F

ret

is used that represents the fraction available for uptake. For leave-on cosmetics

(e.g. creams, body lotion, etc.) mostly a fraction of 1 (100%) is used, while for rinse-off

cosmetics (e.g. shower gel, shampoo, etc.) a smaller fraction is used that depends on the

respective product. In Tables 3A and 3B retention factors are listed that are applied by the

SCCS.

External dermal exposure (E

dermal

) per day for a substance from a certain product category

can be calculated according to:

The daily amount (q

) and retention factor (f

ret x

) are specific to the product category under

consideration, and do not depend on the substance. When multiplied, they yield the daily

effective amount per product category, Eproduct = q

x f

ret x

, which is listed in Tables 3A and

3B for the most important product categories. Multiplied with the concentration or fraction of

a substance in a product, they yield the external dermal exposure to a substance per product

category E

dermal x

as shown in equation (3).

This external exposure can be used to calculate the SED by multiplying with the chemical-

and route-specific uptake rate and normalisation by the bodyweight (see chapter 3-3.5.4).

In cases where the amount per day q

is not given or if more detailed probabilistic

assessments should be performed, the amount per day can be calculated from the frequency

of application (Table 4) and the amount per application. In Appendix 7 (Table A.7) a

literature review can be found listing studies which provide detailed external exposure values

to different cosmetic products. These are given for specific countries. Furthermore, the

external daily exposure per product category can be used to derive a LED (2). Normally, local

dermal effects depend on the surface load, so that the total dermal exposure is normalised

by the Skin Surface Area of application (SSA).

3-3.4.1.2 ORAL EXPOSURE MODELS

The same principles as described for dermal exposure can be applied for oral exposure.

Ingestion can be calculated according to equation (3) by applying adequate retention factors.

Such oral retention factors are needed to take into account that only a fraction of the orally

applied products will be ingested. Since orally applied cosmetics such as toothpaste,

mouthwash or lipstick are normally not intended to be ingested, such retention factors will

normally be small.

dermal x

= C

X q

X f

ret x

(3)

dermal x

(mg/day): external exposure available for dermal uptake from product

Category

EC3 value (%)

Extreme

≤0.2

Strong

>0.2 - ≤ 2

Moderate

Because the guinea pig test methods often do not provide dose-response data, the

intradermal induction concentration in the GPMT and the topical induction concentration in

the Buehler test are used for subcategorisation (ECB, 2002; Basketter et al., 2005). In the

absence of LLNA data, this subcategorisation can be used as indicative for potency.

3-4.7.1 SKIN SENSITISATION QUANTITATIVE RISK ASSESSMENT (QRA)

QRA has been developed for fragrance substances, only. The basic principles of the QRA are

presented in SCCP/1153/08. It is based on the dose of a sensitising chemical, not expected

to cause induction of sensitisation (No Expected Sensitising Induction Level or NESIL), which

may be derived from animal and human data. The NESIL is adjusted by a number of

uncertainty factors (Sensitisation Assessment Factors, SAFs) in order to calculate an

Acceptable Exposure Level (AEL). In addition, a Consumer Exposure Level (CEL) is calculated.

The AEL is then compared with the CEL, whereby, for an acceptable risk, the AEL should be

greater than or equal to the CEL. Within the IDEA project (https://www.ideaproject.info/)

_____________________________________________________________________________________________

the QRA was further refined by including aggregate exposure assessment and revising the

SAFs.

A technical dossier describing the revised QRA (QRA 2) was submitted by the fragrance

industry to the SCCS. After evaluation of the methodology, SCCS concluded that a lot of

progress had been achieved since the initial publication of the QRA. Recently, a peer-reviewed

publication on the QRA2 methodology was published (Api et al., 2020), summarising the

progress made in this field so far. The SCCS considers that it is not yet possible to use the

QRA2 to establish a concentration at which induction of sensitisation of a fragrance is unlikely

to occur. Several aspects of the methodology are not clear and the scientific rationale behind

the methodology needs to be better described. With some revision, this could be a useful

methodology not only for safety evaluation of fragrance allergens, but potentially also for

other cosmetic ingredients. SCCS/1589/17).

In particular, in the case of new substances, post-marketing surveillance would be essential

(see also SCCS/1459/11) to monitor that their use in cosmetics does not lead to allergic

contact dermatitis in consumers, in line with the SCCS Memorandum on use of human data

(SCCS/1576/15).

3-4.7.2 NEXT-GENERATION RISK ASSESSMENT APPROACH (NGRA)

NGRA developed for systemic toxicity of cosmetic ingredients (Berggren et al., 2017, see also

Fig 5 under 3-4.1) has been used as a framework for skin sensitisation safety assessment

(Gilmour et al., 2020), taking the same principles into consideration. A tiered workflow is

applied as illustrated in Fig 7 for skin sensitisation.

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Fig 7: Framework for skin sensitisation safety assessment (Gilmour et al., 2020). S/NS:

safe/not safe; DA: defined approach; WoE; weight of evidence; PoD: point of departure;

RA+/RA-: risk assessment positive/negative outcome. Taken from Regulatory Toxicology and

Pharmacology, 2020, Volume 116, Gilmour et al., with permission from Elsevier.

For the SCCS, NGRA is a novel conceptual approach that offers the possibility to integrate

existing data, read-across information and NAM information in a systematic iterative

approach. Until the NGRA is assessed and accepted to be valid by the SCCS, submissions

including NGRA for skin sensitisation will be evaluated by the SCCS on a case-by-case basis.

The key aspects of NGRA for skin sensitisation can be found in the publication by Gilmour et

al., 2020.

3-4.8 REPEATED DOSE TOXICITY

Repeated dose toxicity studies are performed to investigate toxicological effects (excluding

reproductive, genotoxic and carcinogenic effects) occurring as a result of repeated daily

dosing with, or exposure to, a substance for a specific part of the expected lifespan of the

test species.

A. NAMs

No validated alternative methods are available yet for determining the repeated dose toxicity

of a substance, which poses a problem for the introduction of new compounds e.g.

preservatives on the EU market as this assay usually provides the PoD of the compound under

investigation (necessary for MoS calculation). Efforts are being made by the cosmetic industry

to develop an NGRA strategy as an alternative for not having a PoD generated via in vivo

methodology (see 3-4 and 3-4.1). The topic was extensively discussed in the February 2019

SCCS methodology workshop with the aim to progress from concept to the practical use of

NGRA with a focus on systemic toxicity (Rogiers et al., 2020). Several case studies were

presented and progress has clearly been made, but more case studies and validated NAMs

are needed to create the necessary confidence that NGRA is protective for new compounds

and that unexpected side effects are not occurring.

B. In vivo methods

The following in vivo repeated dose toxicity studies with OECD guidelines are available:

− Sub-acute oral toxicity (28 days) (EC B.7, OECD 407)

− Sub-acute dermal toxicity study (28 days) (EC B.9, OECD 410)

− Sub-acute inhalation toxicity study (28 days) (EC B.8, OECD 412)

− Sub-chronic oral toxicity study: repeated dose 90-day

oral toxicity study in rodents (EC B.26, OECD 408)

− Sub-chronic oral toxicity study: repeated dose 90-day

oral toxicity study in non-rodents (EC B.27, OECD 409)

− Sub-chronic dermal toxicity study: repeated dose 90-day

dermal toxicity study using rodent species (EC B.28, OECD 411)

− Sub-chronic inhalation toxicity study: repeated dose 90-day

inhalation toxicity study using rodent species (EC B.29, OECD 413)

_____________________________________________________________________________________________

− Chronic toxicity studies (primarily rodents) (EC B.30, OECD 452)

− Combined chronic toxicity/carcinogenicity studies (EC B.33, OECD 453)

(primarily rodents)

In the case of the development of cosmetic ingredients that will be in contact with human

skin and mucosae repeatedly, the SCCS is convinced that evaluation of the systemic toxicity

is a key element in safety assessment.

3-4.8.1 THE USE OF UNCERTAINTY FACTORS (UFS) FOR EXTRAPOLATION FOR STUDY DURATION

This type of UF is used to take account of probable differences between the experimental

setting from which the PoD is taken and the human real life situation (use scenario) in case

substance-specific information is lacking.

For some cosmetic ingredients, dermal repeated dose toxicity studies are submitted. These

studies, if of good quality, are taken into consideration by the SCCS as it is the most

commonly used application route for cosmetics. In practice, however, oral route studies are

often used for the MoS calculation to consider (worst case) systemic exposure. Oral repeated

dose toxicity studies can be either subacute (28 days), subchronic (90 days) or chronic

(taking 85% of lifetime).

The 90-day oral toxicity test in rodents was, historically speaking, the most commonly used

repeated dose toxicity test for cosmetic ingredients. Based on the exposure and the short

lifetime of cosmetic products (regularly changing ingredients and concentrations), the 90-day

test provides a good indication of the target organs and the type of systemic toxicity.

In case only a 28-day study is available, the SCCS recommends applying a factor to take

uncertainty into consideration to extrapolate from subacute (28 days) to subchronic (90 days)

toxicity. Different values are being proposed and the choice is made on a case-by-case basis

taking the strengths and weaknesses of the available studies into consideration. The SCCS

commonly uses for such extrapolation a conservative UF of 3. Recently, Escher et al. (2020)

provided data showing that in such a case a factor of 1.5 would be sufficient.

When a scientifically sound 90-day study is available which allows for the determination of a

clear PoD, the SCCS takes this study into account for calculating the MoS. An uncertainty

factor is only included when some doubt exists with respect to the quality of the subchronic

toxicity study and/or in the absence of further information supporting the PoD from the 90-

day study (e.g. availability of a chronic study). Escher et al. (2020) proposed a factor of 1.5.

In other domains (environmental, food, …) higher factors have been proposed, but these may

contribute to a higher variance. In any case, the use of additional UFs needs to be carefully

considered. Indeed, many authors warn that a composite UF may lead to over conservatism

(Simon et al., 2016; Escher et al., 2020). In particular, in the case of aggregate exposure,

using a deterministic exposure assessment multiplication of single UFs may lead to possibly

overly conservative estimates (EFSA, 2012a).

The inhalation route was only rarely used in repeated dose toxicity testing of cosmetic

ingredients due to the lack of relevance for the majority of cosmetic products. This exposure

route is, however, important where a cosmetic ingredient is volatile or a product is intended

to be used in an aerosolised, sprayable or powdered form that could lead to exposure of the

consumer via inhalation. Because of the likelihood of high uncertainty in regard to different

inhalable products and their modes of delivery, the SCCS recommends analysis of uncertainty

on a case-by-case basis).

When reproductive toxicity studies are used to determine the PoD, the uncertainty factors for

extrapolation for study duration are not used.

In sections 3.5.1.1 and 3.5.1.2 a number of default factors are discussed.

_____________________________________________________________________________________________

3-4.8.2 SELECTION OF POD

In repeated dose toxicity studies, the target(s) organ(s) and critical endpoint(s) may be

identified. The critical endpoint is defined as the first (in terms of dose level) adverse effect

associated with the substance. This effect should be biologically relevant for human health

and also in the context of cosmetic exposure. For example, local effects on the gastrointestinal

tract, sometimes observed with irritants after oral exposure, are not considered relevant by

the SCCS to be used for the MoS calculation. A BMD, NOAEL or LOAEL (PoD) is then derived

for each study and the most relevant study in terms of quality, duration of exposure, and

available PoD is then selected by the SCCS to be used for the safety evaluation. If the dose

regimen of a study was limited to 5 days treatment per week, the derived PoD will be

corrected by a factor of 5/7. In analogy, a correction will also be made for longer use

periods.

3-4.9 REPRODUCTIVE TOXICITY

The term "reproductive toxicity" is used to describe the adverse effects induced (by a

substance) on any aspect of mammalian reproduction. It covers all phases of the reproductive

cycle, including impairment of male or female reproductive function or capacity and the

induction of non-heritable adverse effects in the progeny such as death, growth retardation,

structural and functional effects.

A. NAMs

No validated alternative method is yet available for reproductive toxicity that covers all

different phases of the reproductive cycle (JRC 2019, JRC 2020).

Since the field of reproductive toxicity is very complex, it is expected that the various phases

cannot be mimicked using one alternative method and that a battery of tests is needed. Three

alternative methods, restricted to the embryotoxicity area, have been developed:

The Whole Embryo Culture test (WEC)

The MicroMass test (MM)

The Embryonic Stem Cell Test (EST)

The last two tests were considered scientifically valid by the ECVAM Scientific Advisory

Committee (ESAC) for placing a substance into one of the three following categories: non-

embryotoxic, weak/moderate-embryotoxic or strong-embryotoxic. The WEC test is still an

animal test and is considered scientifically valid only for identifying strong embryotoxic

substances (ESAC, 2001).

These three tests might be useful in the CMR strategy for screening out embryotoxic

substances. However, they cannot be used for quantitative risk assessment (Marx-Stoelting

et al., 2009).

The complex endpoint of reproduction toxicity is not covered by the above systems.

Several in vitro methodologies, each covering one of the three biological components of the

reproductive cycle (male and female fertility, implantation and pre- and postnatal

development), were developed under the EU project ReProTect.

The tests reflect various toxicological mechanisms such as effects on Leydig and Sertoli cells,

folliculogenesis, germ cell maturation, motility of sperm cells, steroidogenesis, the endocrine

system, fertilisation, and on the pre-implantation embryo. Neverthless, more information and

research are needed until regulatory acceptance can be envisaged (Schenk et al., 2010).

An extensive review of the actual situation can be found in a JRC report (JRC 2019, JRC

2020). In view of the utmost importance of consumer safety, toxicological evaluation against

some complex endpoints, such as reproductive toxicity, still necessitate the use of animals.

_____________________________________________________________________________________________

B. In vivo methods

The most commonly performed in vivo reproductive toxicity studies are:

1) Two-generation reproductive toxicity study (EC B.35, OECD 416)

2) Prenatal developmental toxicity study

- rodent and non-rodent (EC B.31, OECD 414)

A "Reproduction/Developmental Toxicity Screening Test" (OECD 421) also exsits, as well as

a "Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity

Screening Test" (OECD 422).

The Extended One-Generation Reproductive Toxicity Study (EOGRTS) has been adopted by

the OECD (OECD 443) and a Guidance Document has been established (OECD 2018). It offers

several advantages compared to older OECD TGs and is extensively used:

Compared to OECD TG 416 a significant number of animals can be saved.

More parameters are addressed (e.g. clinical-chemical parameters as in repeated dose

studies; developmental immunotoxicity and neurotoxicity in case such cohorts are

included). Endocrine disruption endpoints are included- (e.g., nipple retention,

anogenital distance at birth, vaginal patency and balanopreputial separation)

Increased statistical power with respect to parameters for reproductive toxicity

Possibility for modification e.g., to include new endpoints for the assessment of

endocrine active chemicals disrupting the Hypothalamus-Pituitary-Gonad (HPG) axis,

the somatotropic axis, the retinoid signalling pathway, the Hypothalamus-Pituitary-

Thyroid (HPT) axis, the vitamin D signalling pathway and the Peroxisome Proliferator-

Activated Receptor (PPAR) signalling pathway

A study report on reproductive toxicity or on prenatal developmental toxicity is in general

only acceptable when it is based on tests that have been carried out before the animal testing

ban or when generated for compliance with other (non-cosmetic) legislative frameworks; see

Appendix 1, section 3 and Appendix 4).

3-4.10 MUTAGENICITY / GENOTOXICITY

3-4.10.1 DEFINITIONS

Mutagenicity: a mutation is defined as a permanent change in the amount or structure of

the genetic material. The terms ‘mutagenic’ and ‘mutagen’ are used for agents giving rise to

an increased occurrence of mutations in populations of cells and/or organisms and applies

both to heritable genetic changes that may be manifested at the phenotypic level and to the

underlying DNA modifications (including specific base pair changes and chromosomal

aberrations).

Germ cell mutations are those that occur during spermatogenesis/ovogenesis and appear in

the egg or sperm (germ cells) and therefore can be passed on to the organism's offspring.

Somatic mutations are those that occur in cells other than the germ cells, and they cannot

be transmitted to the next generation.

Genotoxicity: the more general terms ‘genotoxic’ and ‘genotoxicity’ apply to agents or

conditions that alter the structure, information content, or segregation of DNA, including

those which cause DNA damage by interfering with normal replication processes, or which

alter its replication in a non-physiological manner (temporarily)..

Often also named teratogenicity test

_____________________________________________________________________________________________

3-4.10.2 MECHANISMS

There are several mechanisms that lead to genotoxicity. In general, DNA damage can arise

through either primary or secondary mechanisms (Schins et al., 2007, Magdolenova et al.,

2014, Evans et al., 2017). Primary genotoxicity can be either direct, where there is a direct

interaction of genotoxic agent with DNA, or indirect, where the genotoxic effect is exerted via

intermediate molecules (such as free radicals, as in oxidative stress). Secondary genotoxicity

is driven by oxidative stress arising from inflammation caused by activation/recruitment of

immune cells such as macrophages or neutrophils. Where the evidence suggests indirect

mechanism (oxidative stress), or secondary mechanisms (e.g. inflammation and oxidative

stress caused by overexpression of the immune cells) - a threshold may be derived from the

toxicological data for use in safety assessment.

Based on recommendations of international groups of scientific experts (Dearfield et al.,

2011), and in accord with EFSA (EFSA, 2011a) and the UK Committee on Mutagenicity (COM,

2011), the evaluation of the potential for mutagenicity of a cosmetic substance should include

information on 1) mutagenicity at the gene level, 2) chromosome breakage and/or

rearrangements (clastogenicity), and 3) numerical chromosome aberrations (aneuploidy). For

this task, genotoxicity tests, which measure irreversible mutation endpoints (gene or

chromosome mutations) should be used. Genotoxicity Indicator tests, which measure DNA

damage without taking into account the consequences of this primary damage, can contribute

to the weight of evidence approach but should not be used as stand-alone tests. Finally,

before undertaking any testing, a thorough review should be carried out of all available data

on the substance under assessment.

A. NAMs

(i) In silico methods for genotoxicity and carcinogenicity

- Genotoxic carcinogens (DNA reactive)

As explained in the testing strategy for mutagenicity/genotoxicity (Figure 8, section 3-4.10),

the use of structure-activity relationship based in silico models and read-across can provide

a useful indication of the mutagenic/genotoxic and carcinogenic potential of a cosmetic

ingredient.

The regulatory requirements for testing certain categories of chemicals have led to a large

database on genotoxicity over the past decades, in particular on bacterial reverse mutation

(Ames) test, as well as on in vitro and in vivo micronucleus tests, and chromosomal

aberration. As a result, there is a better understanding of the mechanisms of

mutagenicity/genotoxicity via direct or indirect interaction of chemical substances with the

genetic material, compared to certain other complex toxicity endpoints.

The knowledge deciphered from the available information has indicated that the chemicals

that can cause mutagenic/genotoxic effects through direct interaction with DNA are either

intrinsically electrophilic, or they can be transformed to electrophilic intermediates. However,

some non-electrophilic chemicals may also cause genotoxic effects directly through reaction

with nucleophilic moieties of proteins and nucleic acids or through direct or indirect DNA

alkylation, acylation or adduct formation, or indirectly through generation of reactive

oxyradicals. On the other hand, some chemicals may contain one or more structural alerts

for genotoxicity but may not cause genotoxic effects because of their (higher) molecular

weight, solubility, chemical reactivity, structural geometry, etc. (Plošnik et al., 2016).

The OECD QSAR Toolbox incorporates a number of databases on mutagenicity/genotoxicity

and carcinogenicity that provide a valuable resource for read-across.

- The toolbox on in vitro genotoxicity includes bacterial mutagenicity ISSSTY; ECHA REACH;

OASIS genotoxicity; EFSA pesticide genotoxicity.

- The databases on in vivo genotoxicity include ECHA REACH; ECVAM genotoxicity and

carcinogenicity; EFSA pesticide genotoxicity; ISSMIC Micronucleus; OASIS Micronucleus,

_____________________________________________________________________________________________

Carcinogenic Potency Database (CPDB) (http://toxnet.nlm.nih.gov/cpdb/cpdb.html)

containing data on substances derived from long-term carcinogenicity tests on chemicals in

rats, mice, dogs, hamsters and non-human primates.

- in silico methods (structure-activity based) for the prediction of carcinogenicity of chemical

substances include the open-source tools LAZAR (https://openrisknet.org/e-

infrastructure/services/110/) and (Q)SAR models such Vega (www.vegahub.eu/).

The availability of large amount of data on mutagenicity/genotoxicity and carcinogenicity has

also enabled the identification of key molecular descriptors and structural alerts associated

with mutagenicity/genotoxicity (e.g. Ashby and Tennant, 1988; Benigni and Bossa, 2008;

Plošnik et al., 2016), and the development of several structure-activity based in silico (Q)SAR

models and read-across systems. A number of these systems have been developed using

high quality data and in accordance with the OECD (2014) criteria and were subjected to

stringent assessments to verify their reliability for use in regulatory risk assessments.

- Non-genotoxic carcinogens (DNA-non reactive)

In comparison to genotoxic carcinogens, the identification of Non-Genotoxic Carcinogens

(NGCs) is much more difficult because, unlike the direct or indirect interaction of genotoxic

substances with DNA, the carcinogenic effects of NGCs may manifest from a variety of

different mechanisms, not always relevant to humans, such as:

 Peroxisomal proliferation that may lead to increased cell proliferation or decreased

apoptosis; inhibition of the gap junction intercellular communication, or DNA methylation

 Induction of oxidative stress that may lead to induction of oxidative stress - either due to

increased production of oxyradicals, or decreased cellular antioxidant defences resulting

in DNA damage

 Induction of hormonal imbalance

 Agonistic and antagonistic interaction with Aryl hydrocarbon Receptor (AhR).

NGC are thought to have a safe exposure threshold or dose; thus, their use in society is

permitted provided that the exposure or intake levels do not exceed the threshold. For these

reasons, the in silico methods for the identification of NGCs are based on a limited number of

structural alerts that have so far been identified.

Examples of available in silico systems

As already mentioned in section 3-4.2, the EU project ANTARES has listed the available free-

access and commercial in silico models and tools on the project website (www.antares-

life.eu/).

The notable free access in silico systems for the assessment of mutagenicity/genotoxicity and

carcinogenicity (for which more information is present in APPENDIX 10) include:

 The Danish QSAR database (http://qsar.food.dtu.dk/)

 The OECD QSAR Toolbox (https://qsartoolbox.org/),

 VEGA QSAR platform (www.vegahub.eu/)

 The US-EPA’s Toxicity Estimation Software Tool (T.E.S.T.)

(www.epa.gov/nrmrl/std/qsar/qsar.html)

 Toxtree (http://toxtree.sourceforge.net/) OpenTox for carcinogenicity

(http://apps.ideaconsult.net:8080/ToxPredict)

 Lazar (https://lazar.in-silico.ch/predict)

 OncoLogic (US EPA) (www.epa.gov/oppt/sf/pubs/oncologic.htm)

The list of in silico models/systems is not exhaustive, and the mention of any model/system here does not

constitute an approval of its quality, or recommendation for use by the SCCS.

_____________________________________________________________________________________________

A number of commercial systems are also available for the assessment of potential

mutagenicity/genotoxicity and carcinogenicity. These include QSAR based systems such as

SciQSAR

(SciMatics, Inc.) and TopKat

(Toxicity Prediction by Komputer Assisted

Technology); molecular fragment-based QSAR expert systems such as CASE-Ultra

(Multicase Inc.) and Leadscope

(Leadscope, Inc.); and expert knowledge-based systems

such as Derek Nexus

(Lhasa Ltd.).

Protocols for in silico assessment of genetic toxicity have been described by Hasselgren et al.

(2019) and a number of studies have assessed the reliability of the in silico methods for the

prediction of genotoxicity and carcinogenicity. The results have generally confirmed that a

number of the in silico systems can provide a high degree of reliability for the estimation of

genotoxic potential of chemicals (Serafimova et al., 2010; Bakhtyari et al., 2013;

www.antares-life.eu/files/antares_mutagenicity_qsar2012.pdf). More recently, Honma et al.

(2019) have tested 17 QSAR tools using a proprietary Ames mutagenicity database containing

12140 new chemicals, at least 85% of which were not included in publicly available or

commercial databases and had not been used in QSAR modelling under the Ames/QSAR

International Challenge Project. Their findings indicate that most tools achieved >50%

sensitivity (positive prediction among all Ames positives) and predictive power (accuracy) as

high as 80%, which is almost equivalent to the inter-laboratory reproducibility of the Ames

tests.

These assessments point out to the potential of in silico methods and models to generate

supporting evidence on the potential mutagenicity/genotoxicity of cosmetic ingredients to

support the WoE on their safety in conjunction with other (in vitro) data. As indicated in

section 3.4.2, the estimates derived from in silico models and read-across can provide useful

additional supporting evidence for hazard assessment, especially when the results are

integrated with other sources of evidence (e.g. in vitro data) into an overall weight of evidence

(WoE) for use in risk assessment of cosmetic ingredients.

(ii) From a 3-test in vitro battery to a 2-test in vitro battery:

Evaluation of several databases has demonstrated that an increase in the number of in vitro

tests performed results in an increase of the number of ‘unexpected positives’ while the

number of ‘unexpected negatives’ decreases (Kirkland et al., 2005). The sensitivities of the

2- and 3-test batteries seem quite comparable (Kirkland et al., 2011). Moreover, the

combination of the bacterial reverse mutation test and the in vitro micronucleus test allowed

the detection of all relevant genotoxic carcinogens and in vivo genotoxicants for which data

existed in the databases that were used (Kirkland et al., 2011). Consequently, EFSA and COM

(2011) recommended the use of these 2 tests as a first step in genotoxicity testing. According

to the REACH Regulation and ECHA Guidance (2017), in order to ensure that the necessary

minimum level of information is provided, at least one further test is required in addition to

the gene mutation test in bacteria, namely: an in vitro chromosome aberration test (OECD

TG 473), or an in vitro micronucleus test (OECD TG 487) using mammalian cells. Although in

vitro chromosome aberration test is considered as a possible alternative option to the in vitro

micronucleus test under REACH, it is now generally agreed that these tests are not equivalent

since the in vitro chromosome aberration test is not optimal formeasuring numerical

chromosome aberrations.

In line with this, the SCCS recommends two tests for the base level testing of cosmetic

substances, represented by the following test systems:

Bacterial Reverse Mutation Test (OECD 471) as a test covering gene mutations. Recently,

OECD TG 471 has been revised with CAS reference numbers of strain-specific positive

controls.

In vitro Micronucleus Test (OECD 487) as a test for both structural (clastogenicity) and

numerical (aneugenicity) chromosome aberrations.

The tests should be performed according to the OECD test guidelines.

_____________________________________________________________________________________________

Cells should be exposed to the test substance both in the presence and absence of an

appropriate metabolic activation system. The most commonly used system is a cofactor

supplemented S9-fraction prepared from the livers of rodents (usually rat) treated with

enzyme-inducing agents such as Aroclor 1254 or a combination of phenobarbital and β-

naphthoflavone. The choice and concentration of a metabolic activation system may depend

on the class of chemical being tested. In some cases, it may be appropriate to utilise more

than one activation system. For azo dyes and diazo compounds in the gene mutation test in

bacteria, the use of a reductive metabolic activation system is recommended

(SCCS/1532/14).

In cases where the bacterial reverse mutation test is not optimal for the measurement of

nanoparticles, biocidal compounds and antibiotics, a scientific justification should be given

and a gene mutation test in mammalian cells the Hprt/Xprt (OECD 476), or the thymidine

kinase Tk (OECD 490)} should be performed.

Additionally, when testing nanomaterials, evidence is needed to show that the nanoparticles

were in contact or internalized by the test system and entered in contact with DNA. For further

considerations of particle-related behavior of substances, the Applicants should refer to

SCCS/1611/19: Guidance on the Safety Assessment of Nanomaterials in Cosmetics.

(iii) Novel in vitro approaches in genotoxicity models:

The recommendations and conclusions from the International Workshops on Genotoxicity

Testing (IWGT) (Martus et al., 2020) concerning different methods are supported by the

SCCS:

- The Ames Test:

o critical issues to be considered to bring TG 471 up to date and make it

consistent with other OECD TGs have been identified (Williams et al., 2019;

Levy et al., 2019a and 2019b).

- The Mammalian Cell Gene Mutation Assays:

o In vitro TransGenic Rodent (TGR) mutagenicity assays, once validated, could

be employed for routine mutagenicity assessment, as they have endogenous

metabolic capacity and consequent ability to generate DNA-reactive

metabolites - properties lacking in cell lines frequently employed for in vitro

testing (White et al., 2019);

o In vitro mutagenicity assays based on immortalised cell lines or primary

hepatocytes from the MutaMouse or lacZ Plasmid Mouse are at an advanced

stage of validation;

o The Phosphatidylnositol glycan class A gene (Pig-a) mutagenicity assay is at

an early stage in terms of safety testing and hazard identification (Bemis and

Heflich, 2019);

o The sensitivity of the Mammalian Cell Gene Mutation Assay can be improved

by the use of XRCC1

−/−

/XPA

−/−

TK6 cells (Ibrahim et al., 2020).

- Novel & Emerging in vitro Mammalian Cell Mutagenicity Test Systems:

o genome-wide loss-of-function screening, mutation characterisation by next

generation sequencing, and fluorescence-based mutation detection can be

promising methods (Evans et al., 2019a).

- The 3D Tissues in Genotoxicity Testing (Pfuhler et al., 2020):

o 3D tissue models simulate in vivo-like conditions regarding cell viability,

proliferation, differentiation, morphology, gene and protein expression. They

can complement classical 2D cell culture-based assays;

o 3D tissue-based genotoxicity assays can be used as 2

tier assays to follow-

up on positive results from standard in vitro assays;

o For adoption of a tissue model as a 2

tier assay, ability to detect the full range

of genotoxic damage (leading to mutagenicity, clastogenicity, aneugenicity)

should be demonstrated;

o The 72-hour protocol for the 3D Reconstructed human Skin MicroNucleus assay

(RSMN) has higher sensitivity than the 48-hour protocol;

o The 3D skin comet and MN assays are now sufficiently validated to move

towards individual OECD Test Guidelines, but an independent peer review of

the validation study is still needed.

_____________________________________________________________________________________________

- High Information Content assays:

o adductomics, global transcriptional profiling, error-reduced single-molecule

sequencing, and multiplexed phenotypic profiling are promising tools for

regulatory purposes (Dertinger et al., 2019).

(iv) In vitro models for secondary genotoxicity:

A significant knowledge gap exists in regard to which in vitro system(s) might be appropriate

for assessing secondary (inflammation-driven) genotoxicity (OECD, 2014). Several in vivo-

like in vitro models addressing inflammation driven genotoxicity have been developed,

ranging from a simple conditioned medium approach (e.g. exposing THP-1 derived

macrophages and then transferring the conditioned medium to bronchial cells) to more

complex co-culture models (Evans et al., 2017, 2019b; Åkerlund et al., 2019). The most

advanced models comprising either two or more different cell types co-cultured with immune

cells have been reviewed (Evans et al., 2017) and discussed during the 7th IWGT in Japan

2017 (Pfuhler et al., 2020; Martus et al., 2020). They encompass cell-to-cell interplay, which

promotes intracellular signalling and molecular crosstalk, representing more in vivo-like

conditions.

(v) Outcome of in vitro tests

If the results from both tests are clearly negative in adequately performed tests, it is very

likely that the substance has no mutagenic potential. Likewise, if the results from both tests

are clearly positive, it is very likely that the substance has mutagenic potential. In both

cases further testing is not necessary.

● If one of the two tests is positive, the substance is considered an in vitro mutagen.

Further testing is needed to exclude potential in vivo mutagenicity (and/or

clastogenicity) of the substance under investigation.

A general scheme of mutagenicity testing of cosmetic ingredients is presented in Figure 8.

Additional information on the in vitro testing can be found in COM2011.

Different and potentially contradicting results may be available from the same test when

performed with non-standardized protocols and carried out by different laboratories. In such

cases, expert judgement should be used to evaluate and interpret the data. Further tests

may be necessary to reach an overall conclusion.

Special attention should be given for poorly soluble chemicals. The determination of solubility

in the culture medium prior to the experiment is mandatory. For such substances that are

not cytotoxic at concentrations lower than the lowest insoluble concentration, the highest

concentration analysed in culture medium should produce turbidity or a precipitate visible by

eye or with the aid of an inverted microscope at the end of the treatment with the test

chemical. Even if cytotoxicity occurs above the lowest insoluble concentration, it is advisable

to test at only one concentration producing turbidity or a visible precipitate because

inaccurate effects may result from the precipitate. At the concentration producing a

precipitate, care should be taken to ensure that the precipitate does not interfere with the

conduct of the test (e.g. staining or scoring).

(vi) Toolbox for further evaluation in a WoE approach

● The comet assay in mammalian cells or on 3D reconstructed human skin can support

a WoE approach in the case of a positive or equivocal bacterial or mammalian gene

mutation test. In June 2020, the 3D reconstructed human skin comet assay has been

presubmitted to EURL ECVAM for assessment. The enzyme-linked comet assay for

detection of oxidized DNA bases can be useful for identification of a genotoxicity

involving oxidative stress. Standardisation and pre-validation of the method have been

conducted recently by the hCOMET consortium (Møller et al., 2020) and the application

for an OECD test guideline is in preparation.

_____________________________________________________________________________________________

● To evaluate a positive or equivocal result, RSMN could be considered for dermally

applied compounds. The experimental phase of the validation has been finalised

(Phfuhler et al., 2020) and the RSMN has been pre-submitted to EURL ECVAM for

assessment. Another tool is the Hen’s Egg test for Micronucleus Induction (HET-MN)

which is currently under evaluation (JRC 2019, 2020; Reisinger et al., 2019).

Negative results from these alternative tests alone might not be sufficient to overrule the

positive results from a recommended test.

● Mechanistic investigations (e.g. toxicogenomics) or internal exposure (toxicokinetics)

are tools that may be helpful in a WoE evaluation. Reporter gene assays based on

human, animal or bacterial cells are tools supporting a WoE approach. Among such

tests are the Green Screen HC™ used to screen the genotoxic and cytotoxic potential

of chemicals and ToxTracker

, which when combined with Vitotox (a mutagenicity

test that can be used as a surrogate for an Ames test) showed a better performance

than observed in the official 2-test battery (Ates et al., 2016). ToxTracker

was able

to accurately classify compounds as genotoxic or non-genotoxic, and could

discriminate between DNA-reactive compounds, aneugens and indirect genotoxicity

caused by oxidative stress (Brandsma et al., 2020).

● The results obtained using a reporter gene assay provide mechanistic information at

the molecular level but cannot alone overrule a positive result from an in vitro battery

as the assay is based on a limited number of genes.

● Another tool to potentially address a positive result in a 2-test battery (in one of the

two assays) is transcriptomics analysis in TK6 cells (Li et al., 2015), HepG2 cells

(Maghoufopoukou et al., 2012) or HepaRG™ cells (Ates et al., 2018), in which a higher

number of genes provide mechanistic information (Dertinger et al., 2019). The level

of phosphorylated form of H2AX histone (γH2AX) in cells exposed to a chemical can

indicate its potential for induction of DNA damage (Kopp et al., 2019). Assays that

simultaneously analyse different biomarkers (e.g., p53, γH2AX, phospho-histone H3

or polyploidy) are being developed to provide mechanistic information on the types of

biological damage induced by different classes of substances. Such promising assays

are MultiFlow and the Multi-Endpoint Genotoxicity Assay (MEGA-Screen system)

(Dertinger et al., 2019).

Despite the possibilities offered by the toolbox, expert judgement may be necessary to be

able to come to an overall conclusion.

Intensive work is being carried out on adapting current tests to high-throughput technologies

(e.g., micronucleus test, Comet assay, yH2AX assay, high content analysis and other assays)

(Collins et al., 2017).

Alternative tests for which no OECD test guideline is currently available should be performed

according to the general principles laid down in OECD test Guidelines (OECD 211).

In cases where a clear positive result cannot be overruled in a WoE approach even with

additional testing, the substance has to be considered a mutagen. A positive in vitro result in

genotoxicity testing is also seen as indicative for the carcinogenic potential of substances.

The SCCS has published an Addendum to the NoG (SCCS/1501/12), in which details such as

definitions, critical steps, crucial experimental conditions to be followed, etc. are described

(SCCS/1532/14).

_____________________________________________________________________________________________

Figure 8. Scheme of testing strategy for genotoxicity/mutagenicity of cosmetic ingredients

_____________________________________________________________________________________________

B. In vivo methods

Animal studies on mutagenicity or genotoxicity are acceptable when data are already

available from tests that have been carried out before the animal testing ban or when

generated for compliance with other legislative (non-cosmetic) frameworks (see Section 1).

When there is a positive result from an in vitro gene mutation test, adequate somatic cell in

vivo tests are:

-a Transgenic Rodent and Germ cell gene mutation assay (TGR) (OECD TG 488),

-an in vivo mammalian alkaline comet assay (OECD TG 489).

It is no longer recommended to perform an Unscheduled DNA Synthesis (UDS) test with

mammalian liver cells in vivo (OECD TG 486) (EFSA, 2017b).

Adequate somatic cell in vivo tests to investigate structural or numerical chromosome

aberrations are:

-a mammalian erythrocyte micronucleus test (OECD TG 474),

-a mammalian bone marrow chromosome aberration test (OECD TG 475)

-an in vivo alkaline comet assay (OECD TG 489).

An OECD guideline for the Pig-a in vivo assay is in progress. According to the experts from

the 7

IWGT, the assay can be valuable as a follow-up to in vitro positive results (Kirkland et

al., 2019).

EFSA concluded that target tissue exposure in in vivo studies should be demonstrated,

particularly in the bone marrow (e.g., mammalian erythrocyte micronucleus assay). Toxicity

to the bone marrow in itself provides sufficient evidence to allow concluding on the validity

of a negative outcome of a study. All other direct or indirect evidences of target tissue

exposure should be assessed within a weight-of-evidence approach.

The SCCS is aware of work being conducted in the development of new generation framework

for assessment of genomic damage (Steiblen et al., 2020; Luijten et al., 2020), however this

work is at preliminary stage and no guidance can be delineated at the moment.

3-4.11 CARCINOGENICITY

Substances are defined as carcinogenic if, after inhalation, ingestion, dermal application or

injection, they induce or increase the incidence of tumours, induce malignancy or shorten the

time before tumour occurrence (ECHA 2017).

Carcinogens are often differentiated as "genotoxic carcinogens" (DNA-reactive substances),

for which the most plausible mode of carcinogenic action is via genotoxic effects (i.e. point

mutations and structural chromosomal aberrations), and "non-genotoxic carcinogens", or

non-DNA reactive substances that are carcinogenic due to mechanisms other than direct

interactions with DNA (ECHA 2017).

A. NAMs

(i) In silico methods for carcinogenicity:

See under 3-4.10.2 (i): in silico methods for genotoxicty and carcinogenicity

_____________________________________________________________________________________________

(ii) In vitro methods

- Genotoxic carcinogens (DNA reactive)

At present validated alternative in vitro methods to determine the carcinogenic potential of

substances are not available as OECD test Guidelines. However, there are new in vitro

approaches which may be helpful in an overall WoE approach to indicate potential genotoxic

as well as NGC substances.

For genotoxic substances, in vitro mutagenicity tests are well developed. Due to the relation

between mutations and cancer, these genotoxicity tests can also be seen as a pre-screening

for carcinogenicity. A positive result in one of the in vitro mutagenicity/ genotoxicity testing

battery may be indicative for considering a substance as a putative carcinogen. This indication

may be further supported by a positive result in Cell Transformation Assays (CTAs, Guidance

documents No 214 and No 231).

Worldwide research is ongoing with regard to in vitro toxicogenomics for the detection of

mutagens, genotoxic carcinogens, and particularly NGC. By global gene expression profiling

via microarray technology, gene patterns covering diverse mechanisms of substance-induced

genotoxicity can be identified (Schmitz-Spanke, 2019).

These gene patterns/biomarkers can be further used as a follow-up of positive findings of the

standard in vitro mutagenicity/genotoxicity testing battery (Goodsaid et al., 2010; Doktorova

et al., 2012; Magkoufopoulou et al., 2012; Ates et al. 2018). In addition to in vitro

mutagenicity/genotoxicity tests (see above), data from in vitro tests combined with

toxicogenomics may also be considered in a WoE approach. A multiple-endpoint approach is

most probably a more reliable means of assessing carcinogenicity in vitro than traditional,

single-endpoint tests (Wilde et al., 2018).

- Non-genotoxic carcinogens (DNA-non reactive)

Genotoxic carcinogens either induce mutations in (short term) eukaryotic and prokaryotic

mutation assays or induce direct DNA damage in the target organ. Although it has been

estimated that 10-20% of recognised human carcinogens classified as Class 1 by IARC act

through NGC mechanisms (Hernandez et al., 2009), there are no specific requirements to

obtain information on NGC mechanisms of carcinogenicity. As such many NGC will remain

unidentified, and as a consequence their risks to human health will not be managed. The

overview of NGC mechanisms presented by Jacobs et al. (Jacobs et al., 2016) indicates that

assays with endpoints capturing early key event mechanisms may provide an individual

contribution to the WoE approach of NGC.

(iii) Development of integrated approach to testing and assessment (IATA) for NGC

Using the AOP concept, an OECD expert working group has elaborated a preliminary panel of

key hallmarks of NGC and representative international standardised tests that can address

IATA for NGC (Jacobs et al., 2020). Using a systematic review approach combined with assay

database mining, overall more than 100 in vitro assays have been identified so far, within 13

cancer hallmark assay blocks that address early, mid and later key events with consequent

increasing associations with adverse outcome. The assays are currently undergoing

evaluation by the group including assessment of their readiness for validation in the short,

medium and long term.

(iv) Cell Transformation Assays (CTA) as a possible alternative to animal models of

carcinogenicity testing

CTA can detect both genotoxic and NGC (Sasaki et al., 2014) and are able to highlight various

stages from early (initiation) to late (promotion) phases (OECD 2017, Serra et al., 2019,

Jacobs et al., 2020). They address several endpoints. They measure cell transformation,

which includes early key events such as transdifferentiation, acquisition of a peculiar

morphology, etc., reflecting stages in the multistep cancer process (for more information,

see Appendix 8, Table A.8). CTAs thus can be used as phenotypic anchoring for mechanistic

_____________________________________________________________________________________________

studies (Callegaro et al, 2017). They may provide additional information and may be used as

a follow-up for confirmation of in vitro positive results from genotoxicity assays, typically as

part of a WoE approach (Doktorova et al., 2012, Creton et al., 2012). When employed in

combination with other information, such as genotoxicity data, structure–activity analysis

and pharmaco/toxicokinetic information, CTAs could facilitate a relatively comprehensive

assessment of carcinogenic potential (Creton et al., 2012, Corvi et al., 2017, Mascolo et al.,

2018). Toxicogenomics in combination with in vitro CTAs allow the identification of the

transcriptionally activated pathways (Mascolo et al., 2018). This integrated approach has the

potential to be considered as part of an IATA for non-genotoxic carcinogenesis (Corvi et al.,

2017).

Validated CTAs are the BALB/c 3T3 CTA (EURL ECVAM, 2012), the Syrian Hamster Embryo

(SHE) CTA OECD Guidance Document No. 214 (OECD, 2015, Corvi et al., 2017) and the Bhas

42 CTA OECD Guidance Document No. 231 (OECD, 2017, Jacobs et al., 2020). These can be

used in a WoE approach in the testing of substances for carcinogenic potential. At present,

the carcinogenic potential of a substance cannot be derived from a stand-alone CTA.

B. In vivo methods

An in vivo carcinogenicity study is only acceptable by SCCS when based on tests that have

been carried out before the animal testing ban or when carried out for the purpose of

compliance with other (non-cosmetic) legislative frameworks.

Usually the carcinogenic potential of a substance is assessed using a 2-year bioassay (OECD

451: carcinogenicity studies). A combined chronic toxicity/carcinogenicity study can also be

performed to identify carcinogenic and the majority of chronic effects, and to determine dose-

response relationships following prolonged and repeated exposure (OECD 453: C-combined

chronic toxicity/carcinogenicity studies). It is now well recognised by the scientific and

regulatory community that the use of the rodent cancer bioassay has many limitations in

terms of reliability and relevance (Jacobs et al., 2020).

Utilising a mode of action analysis instead of performing the long-term rodent carcinogenicity

studies offers a more direct and rational basis for human cancer risk assessment. Such

analysis should be performed whenever possible, rather than simple hazard identification

(Berry 2018, Goodman 2018).

3-4.12 PHOTO-INDUCED TOXICITY

3-4.12.1 PHOTO-IRRITATION AND PHOTO-SENSITISATION

A. NAMs

The "3T3 Neutral Red Uptake Photo-toxicity Test (3T3 NRU PT)" is a validated in vitro method

(EC B.41, OECD 432), based on a comparison of the cytotoxicity of a chemical when tested

in the presence and in the absence of exposure to a non-cytotoxic dose of UV/VIS radiation.

Its use is mandatory for testing for phototoxic potential. It is not designed to predict other

adverse effects that may arise from combined actions of a chemical and light, e.g. it does not

address photoclastogenicity/ photomutagenicity, photo-allergy or photocarcinogenicity.

In the OECD 432 GD it is indicated that if the Molar Extinction/absorption Coefficient (MEC)

is less than 1000 L mol⁻¹ cm⁻¹ (measured in methanol), the chemical is unlikely to be

photoreactive and that such chemicals may not need to be tested.

EFSA (2016) concluded that for a light source emitting wavelengths mainly below 320 nm,

more guidance is needed on how to interpret the data and on how to perform the test with a

light source emitting between 290 and 320 nm. In the OECD TG, it is mentioned that

cytotoxicity increases 1000-fold as the wavelength ranges from 313 to 280 nm. Although the

data requirement in Reg. (EU) No. 283/2013 are for substances absorbing electromagnetic

radiation in the wavelength range 290-700 nm, there are difficulties in testing below 320 nm.

_____________________________________________________________________________________________

EFSA proposed that the phototoxicity test should not be performed if it has been

demonstrated that the test material only absorbs at wavelengths lower than 313 nm and if

there is insufficient absorption at longer wavelengths.

As a second tier, the biological effects can be further evaluated on a reconstructed human

skin model with some barrier properties (Kandarova, 2011). A positive control should always

be included. A negative result for the compound under consideration is usually accepted. To

enhance the chance of achieving correctly predicted results of phototoxic potential of

chemicals, a more complex screening using UV/VIS radiation spectral analysis and Reactive

Oxygen Species (ROS)/micellar ROS (mROS) assays could be used according to Nishida et

al., 2015.

Presently, no validated in vitro methods for the detection of photo-sensitisation are available.

Nevertheless, it is expected that chemicals showing photo-allergic properties are likely to give

positive reactions in the 3T3 NRU PT test. There is also work being conducted on some other

in vitro tests for photo-allergenic potential such as: photo-hCLAT, NCTC2455 assay, dendritic

cell-based assay, or photo-SH/NH2 test (Onoue et al., 2017).

For pharmaceuticals applied to the skin, it is stated in EMA 2012 (updated 2015) that

reconstructed human skin models can be used. Under adequate test conditions, a negative

result in a reconstructed human skin assay indicates that the direct phototoxicity potential of

the formulation can be regarded as low. In that case, generally no further phototoxicity

testing is recommended.

B. In vivo methods

At present, no official guideline-based protocols for photo-irritation and photo-sensitisation

testing in animals have been evaluated. Several industry reports describe test protocols. For

pharmaceuticals, guidance on such testing is available (FDA, 2015; EMA, 2012). These

documents do not, however, specify protocols for the testing of adverse effects of orally or

topically applied agents, nor do they give recommendations about the species to be used.

The SCCS guidance is as follows:

UV-VIS spectra of the compound along with the MEC, determined according to a harmonised

procedure, should be provided.

There is no requirement for phototoxicity testing of compounds with a MEC below 1000 L

mol−1 cm−1.

There is no requirement for in vitro phototoxicity testing if the test material only absorbs at

wavelengths lower than 313 nm and if there is insufficient absorption at longer wavelengths.

3-4.12.2 PHOTOMUTAGENICITY / PHOTOGENOTOXICITY

Photomutagenic or photogenotoxic chemicals are chemicals that absorb visible (VIS) light or

UV radiation and, through activation to a more reactive state or release of free radicals, cause

damage to DNA and induce gene mutations or chromosome aberrations.

The terms ‘‘photomutagenesis’’ or ‘‘photogenotoxicity’’ are used to describe the ‘indirect’

induction of gene mutations or chromosomal aberrations after transfer of energy or charge

from a light absorbing molecule other than DNA (Müller and Gocke, 2013). This includes the

genotoxic effects elicited by degradation products and/or radicals generated by VIS and UV

wavelengths.

(i) Current status of tests available for photogenotoxicity/photomutagenicity

assessment

A previous version of the Notes of Guidance (SCCNFP/0690/03) already mentioned that for

the detection of photochemical clastogenicity/mutagenicity, several assays had been adapted

_____________________________________________________________________________________________

to a combined treatment of chemicals with UV-VIS radiation (Averbeck et al., 1979; Dean et

al., 1991; Chetelat et al., 1993a,b, 1996; Gocke et al., 1998; Pflaum et al., 1998; Kersten et

al., 2002).

The existing principles and test methods in the field of photomutagenicity/photogenotoxicity

was summarised in the report of the Gesellschaft für Umweltmutationsforschung (GUM) Task

Force on photochemical genotoxicity (Brendler-Schwaab et al., 2004). The methods described

include the photo-Ames test, the photo HPRT/photomouse lymphoma assay, the photo-

micronucleus test, the photochromosome aberration test and the photo-Comet assay. In

many cases, the concurrent use of irradiation, while performing a standard

mutagenicity/genotoxicity study, does not significantly alter the existing OECD protocol

without irradiation. Therefore, the majority of the described photomutagenicity/photo-

genotoxicity tests are considered as being valid.

In addition to the conclusions of an international workshop (Lynch et al., 2011), a

comprehensive review (Müller and Gocke, 2013) concluded that “photomutagenicity is not

suitable for a general testing framework within cosmetic or pharmaceutical testing guidelines”

and suggested a case-by-case approach

(ii) Guidances for photogenotoxicity/photomutagenicity testing

The COM (COM 2013) recommended that photogenotoxicity testing does not need to be

undertaken routinely as part of a photosafety assessment and that photogenotoxicity testing

had a negligible impact in the overall assessment for potential of photocarcinogenicity.

Moreover, if there is a negative response from the phototoxicity test, no photomutagenicity

test is required. However, if the test is positive, no specific guidance is provided.

The International Conference on Harmonisation (ICH) guideline on photosafety evaluation of

pharmaceuticals (Step 4 of the ICH Process dated 13 November 2013) stated: ‘Note 2. Testing

for photogenotoxicity is not recommended as a part of the standard photosafety testing

program as in most cases, the mechanism by which compounds induce photogenotoxic effects

is identical to those that produce phototoxicity, and thus separate testing of both endpoints

is not warranted.’

The ICH guideline has been adopted in EU by the Committee for Medicinal Products for Human

use (CHMP) in December 2015 and issued as EMA/CHMP/ICH/752211/2012 (EMA, 2015) as

well as in the USA by the FDA and issued as FDA/2013/D/0068 (FDA, 2015).

In 2016 the EFSA (2016) agreed that photomutagenicity testing is not required for the time

being, unless further guidance is provided. Additionally, they concluded that the concern

regarding positive results in the phototoxicity test should be raised to the risk managers in

the conclusion of the peer review.

In this regard, taking also into consideration the general recommendations regarding the

experimental conduct of tests for photogenotoxicity (Gocke et al., 2000), the SCCS

guidance is as follows:

- although the validity of photomutagenicity/photogenotoxicity testing is being questioned,

in specific cases when the structure of a molecule, its light absorbing potential or its

potential to be photo-activated may indicate a photomutagenic/photogenotoxic hazard,

then photomutagenicity tests should be provided, including gene mutations and

clastogenicity/aneugenicity endpoints; especially when the substance is liable to reach the

eyes or light-exposed areas of skin, either by direct contact or through systemic

distribution. Additionally, available alternative methods, for example scientifically

validated comet assay for detection of oxidized DNA lesions, or in silico methods, can be

considered.

- UV-VIS spectra of the compound along with the MEC, determined according to a

harmonised procedure, should be provided.

- the phototoxicity test should not be performed if the test material only absorbs at

wavelengths lower than 313 nm and if there is insufficient absorption at longer

wavelengths.

- no photomutagenicity tests are needed when the phototoxicity tests are negative.

_____________________________________________________________________________________________

- there is no requirement for a photomutagenicity test if the test material only absorbs at

wavelengths lower than 313 nm and if there is insufficient absorption at longer

wavelengths.

3-4.13 HUMAN DATA IN HAZARD ASSESSMENT

Tests in animals and alternative methods may have limited predictive value with respect to

the human situation. Therefore, when human data is available, this information is very

valuable. Human data can be obtained via various sources. For bioavailability and systemic

toxicology information, sources could be: post-marketing surveillance data, results from

biomonitoring programs (see Section 3-3.5.6), case reports, occupational surveillance data

and occupational disease registries (e.g. from production of the ingredient or when the

cosmetic ingredient is also used in non-cosmetic areas), poison centre information,

epidemiological studies, clinical studies, tests with human volunteers.

Tests with human volunteers (e.g. skin compatibility tests) confirm that there are no harmful

effects when applying a cosmetic product for the first time to human skin or mucous

membranes. If considered scientifically and ethically necessary, human tests can only be

envisaged, provided that the toxicological profiles of the components are available and no

concern is raised. A high degree of safety needs to be ensured. Finished cosmetic products

are usually tested in a small group of human volunteers to confirm skin and mucous

membrane compatibility, as well as cosmetic acceptability (fulfilment of in-use expectations).

Human studies might also become necessary to build up and validate PBPK models (see

Section 3-3.5.3).

The general ethical and practical aspects related to human volunteer compatibility studies on

finished cosmetic products, are described in SCCNFP/0068/98 (for skin irritancy) and

SCCNFP/0245/99 (for skin sensitisation). For skin sensitisation, human patch test data, if

available, have to be taken into account (SCCS/1567/15).

Predictive testing of potentially skin sensitising cosmetic (mixtures of) substances

(SCCNFP/0120/99) is more controversial than the irritancy testing, since these tests carry

the risk of inducing a long-lasting or permanent immunological sensitisation in the individual.

Therefore, serious ethical questions arise.

Despite many years of experience with human sensitisation tests, limited scientific

information is available regarding the consequences involved for human volunteers who have

developed sensitisation as a result of such testing.

Due to the uncertainties mentioned, the SCCS is of the opinion that predictive human

sensitisation tests should not be carried out.

The same ethical restrictions apply to human predictive tests on photosensitisation. For

photosensitisation, information can be obtained from published clinical studies and case

reports. There are no officially adopted guidelines or protocols, but in general the test

procedures are quite similar to those used in photo-patch testing in clinical settings

(Bruynzeel, 2004). Normally a UV-A dose of 5 – 10 J (and occasionally UV-B in appropriate

non-erythemogenic dose) is applied to a skin area that has been exposed to the product or

substance during the preceding 24 hours. Adequate control test areas, including a vehicle

exposed and an unexposed UV irradiated area, are essential. Readings must be performed at

least at 4, 24 and 48 hours after irradiation.

3-4.14 OTHER CONSIDERATIONS

When HBM in used in the safety evaluation of consumer product ingredients, the following

limitations apply:

_____________________________________________________________________________________________

− HBM is applicable to substances that are systemically taken up and where the half-life

of the biomarker enables sampling and analytical determination.

− HBM is not appropriate when the relevant biomarker is an endogenously formed

substance, present in much higher concentrations than those caused by the uptake of

a substance from the environment or consumer products.

− HBM is not appropriate when the relevant biomarker is non-specific (e.g., can be

formed by different parent compounds such as hippuric acid).

− Various factors influence HBM results, including age, gender, lifestyle, consumer

habits, diet, place of residence, etc., as they modify the amounts of chemical

substances taken up. Inter-individual differences in the metabolism of chemical

substances, excretion of metabolites, health status as well as different compositions

of biological materials like varying dilutions of urine etc., even under identical

conditions of exposure, may provide different HBM results.

− Other error sources are contamination of samples during collection and handling of

the biological samples (Calafat and Needham, 2009).

3-5 GENERAL PRINCIPLES FOR THE CALCULATION OF THE MARGIN OF SAFETY AND

THRESHOLD OF TOXICOLOGICAL CONCERN

3-5.1 CALCULATION OF THE MARGIN OF SAFETY OF A COSMETIC INGREDIENT

The last step in the safety evaluation of a cosmetic ingredient is the calculation of the MoS,

which is the ratio between a PoD

sys

(usually historical NOAEL or BMD values from oral studies)

and an estimate of the exposure (11).

Mostly, only a repeated dose toxicity study with oral exposure is available as a surrogate for

a study with dermal exposure. For comparison with the PoD

sys

, usually an SED for the dermal

route is derived as the exposure estimate. For calculation of SED, see 3-3.5.4.

Where possible, a BMD is used as PoD

sys

{see also 3-1 (3)}.

PoD

sys

MoS = (11)

SED

3-5.1.1 THE POD VALUE

As far as the determination of critical effects in repeated dose toxicity studies is concerned,

the available repeated dose toxicity data should be evaluated in detail for characterisation of

the health hazards upon repeated exposure. In this process, an assessment of all toxicological

effect(s), their dose-response relationships and possible thresholds should be taken into

account. The evaluation should include an assessment of the severity of the effect(s), whether

the observed effect(s) are adverse or adaptive, irreversible or not - and whether they are

precursors or not of significant effects or secondary to general toxicity. Correlations between

changes in several parameters (e.g. between clinical or biochemical measurements, organ

weights and (histo)pathological effects) will be helpful in the evaluation of the nature of the

effects. Further guidance on this issue can be found in several publications (WHO, 1994;

WHO, 1999; ECETOC, 2002; ECHA, 2012a).

3-5.1.1.1 DETERMINATION OF NOAEL

The NOAEL is defined as the highest dose or exposure level where no (adverse)

treatment-related findings are observed. For cosmetic ingredients, the NOAEL is mainly

_____________________________________________________________________________________________

derived from a 90-day repeated dose animal study or from a reproductive toxicity

animal study.

The BMD approach should preferentially be used as the dose descriptor for the PoD and the

MoS calculations (EFSA, 2009). When no BMD can be calculated, usually historical NOAEL

values are applied.

If a BMD or a NOAEL cannot be identified from the available data, other dose descriptors

such as the Lowest Observed (Adverse) Effect Level (LOAEL) may be used in the MoS

calculation.

See Section 3-1(3)(4).

3-5.1.1.2 DETERMINATION OF BMD

Although not limited to in vivo data, it involves first fitting a dose-response model to the data

and then interpolating to find the lowest dose that causes a statistically significant response

(or alternatively: the dose that corresponds to a low but measurable change in response

over the entire dose interval). That dose is defined as the BMD. To account for uncertainty,

a two-sided 90% confidence interval for the BMD interval, the BMDU (upper confidence limit

of BMD), is sometimes used to calculate the BMDU/BMDL (lower confidence limit of BMD)

ratio which provides an estimate of the uncertainty in the BMD value. The BMD/BMDL ratio

can also be used for this purpose but is less suitable as it is does not take the full uncertainty

in the BMD estimation into account (EFSA guidance, 2017c).

With quantal data, also referred to as dose-response data, the outcomes are incidences, e.g.

number and gender of animals with signs of toxicity. With such data the BMD is defined as

the dose associated with a specific change in the response, the Benchmark Response (BMR)

most often defined as either an increased additional risk or extra risk. An extra risk of 10%

is recommended as default for the BMR by both EFSA (EFSA, 2016) and US EPA (US EPA,

2010).

Body weight, organ weights and enzyme levels are typical continuous data, also referred to

as dose-effect data. For such data each animal has its own magnitude of effect and the

arithmetic or geometric means of the different dose groups are usually compared.

EFSA has proposed a preferred default 5% as a BMR, with modifications if required by

toxicological or statistical considerations (EFSA, 2017c).

3-5.1.1.3 CHOICE OF MODELS

The most well-known BMD software (BMDS) has been developed by the US EPA

(www.epa.gov/bmds) and the National Institute for Public Health and the Environment

(RIVM) (the PROAST software, www.rivm.nl/proast). Application of different models to the

same data will yield different values for the BMD and BMDL. As a consequence, there are

different methods that guide the choice of which BMD and BMDL to use. Current EFSA

guidelines suggest that the lowest BMDL among the models that pass a goodness-of-fit test

should be used as the PoD (EFSA, 2017c). EPA’s guidelines are less conservative, suggesting

that the model with the lowest Akaike Information Criterion (AIC) should be used as the PoD,

unless there is a large difference between the BMDL values obtained with the different models

(US EPA, 2012).

The AIC takes the likelihood of the model fit into account, but penalizes models with many

parameters:

The SCCS considers that there are still practical considerations regarding the use of this

approach when evaluating cosmetic ingredients and its application requires a level of expert

judgement and modelling expertise.

_____________________________________________________________________________________________

3-5.1.1.4 ADJUSTMENT FACTORS TO THE POD

Dependent on dosing regimen, adjustment to daily exposure should be performed. For

example, if the dose regimen in such a study was only 5 days treatment per week, a PoD

corrected by a factor of 5/7 should be used for the MoS calculation (ECHA, 2012a).

When the PoD is based on a LOAEL, often an additional assessment factor of 3 is

added in the calculation of the MoS. However, a higher assessment factor of up to 10

may be decided on a case-by-case basis, taking into account the dose spacing in the

performed repeated dose toxicity test, the shape and slope of the dose-response curve (and

in some cases the extent and severity of the effect(s) seen when LOAEL values are used). In

some cases, the study cannot be used for safety assessment.

In case a 90-day repeated dose toxicity study is not available, a NOAEL or BMDL from a 28-

day repeated dose toxicity study can be used in the MoS calculation for a cosmetic ingredient.

In this case, a default assessment factor of 3 for exposure duration may be used in the

calculation of the MoS.

3-5.1.2 THE PODSYS VALUE

If the absorption by the oral route is 100%, then the external and internal doses of the oral

route are the same. If the absorption by the oral route is less than 100%, which is often the

case, the procedure may underestimate the risk of the exposure of the non-oral route.

It is considered that not more than 50% of an orally administered dose is systemically

available. Thus, in the absence of data, 50% of the administered dose is used as the

default oral absorption value for a cosmetic ingredient and the PoDsys is derived from the

PoD by dividing with a factor 2. If there is information to suggest poor oral bioavailability,

a default value of 10% oral absorption could be considered. However, whenever oral

absorption data are available, these should be used, also when using other dose descriptors.

Also, any other available kinetic data should be considered.

For chemicals with a high first-pass metabolism in the gut or liver, the situation is even more

complex and, in addition, the target organ for toxicity has to be taken into consideration and

route-to-route extrapolation may not be adequate.

In the case of oral to inhalation extrapolation, a default factor of 2

is also proposed

(default absorption oral route: 50%; inhalation 100%; ECHA, 2012a).

3-5.1.3 MOS ANALYSIS

The calculated MoS is compared with a reference MoS, which is comparable to the

uncertainty/assessment factor used in risk and safety assessments to extrapolate from a

group of test animals to an average human being, and subsequently from average humans

to sensitive subpopulations (see Figure 9). A default value of 100 (10x10) accounting for

inter- and intraspecies differences is generally accepted and a MoS of at least 100 therefore

indicates that a cosmetic ingredient is considered safe for use.

Besides the default value of 50% for oral absorption, in this guidance, another default value of 50% for dermal

absorption should be distinguished if no adequate dermal absorption data is available {see Section 3-3.5.2}.

_____________________________________________________________________________________________

Figure 9: Schematic representation of the extrapolation from animal to man

(Renwick, 1998).

As shown in Figure 9, the default value of 100 consists of a factor of 10 for the extrapolation

from test animals to an average human being (interspecies extrapolation) and another factor

of 10 taking into account the variations within the human population (intra-species

extrapolation). These factors can be further subdivided as indicated in Figure 10.

When considerable qualitative/quantitative toxicokinetic differences are observed between

test animals and humans, as well as within human individuals, e.g. from relevant toxicokinetic

data for rat and/or humans (SCCS/1443/11, SCCS/1479/12), the interspecies and/or intra-

species toxicokinetic default factor (see Figure 10) can be increased/decreased (case-by-

case evaluation).

Regarding substance-specific information for variations in toxicodynamics, deviation from the

default value is possible if sufficiently justified. For instance, in case of different susceptibility

to HPT-axis disturbances in rats and humans, a change of the interspecies toxicodynamic

default factor of 2.5 may be required (SCCS/1481/12).

Figure 10: Further subdivision of the uncertainty/assessment factor, taking toxicokinetics

and toxicodynamics into account (based on WHO, 1994).

* including historical NOAEL values

_____________________________________________________________________________________________

Additional considerations:

i. Some cosmetic substances are not used on a daily basis, although their NOAEL values

have been obtained in studies after daily administration of the substances. Combining

these NOAEL values with daily exposures therefore results in a clear overestimation of the

risk. The comparison of a NOAEL resulting from a daily exposure study with the SED of a

certain cosmetic ingredient is therefore accepted as a conservative estimate, even if it is

only applied e.g. once per week or once per month. However, the daily amount for product

categories with low frequencies of application may not be adjusted by the frequency (i.e.

not divided by 30, if applied once per month), as justified by: "The actual daily dose is

independent of the exposure frequency. This means that if, for a certain scenario, worker

or consumer exposure is only for a number of days per year, the exposure value is the

actual dose on the exposure days, and not the daily dose averaged out (and thus divided!)

over the whole year" (ECHA, 2012a). This reasoning, however, may be changed for

example in the case of hair dyes (e.g. oxidative hair dyes only applied once per month)

and a MoS slightly below 100. One could consider a substance as being safe due to the

occasional use and the built-in conservatism of assessment but only after expert

judgement.

ii. When there is sufficient evidence that the dermal absorption of a cosmetic ingredient is

very low, systemic exposure may be negligible and the calculation of a MoS may not be

justified or applicable (see Sections 3-6.11 and 3-5.2). See also for example UV filter

HAA299 SCCS/1533/14.

iii. The SCCS will decide upon the relevance of MoS calculations on a case-by-case basis,

taking into account the general toxicological profile of the substance under consideration,

its toxicokinetic properties and its intended use.

iv. With regard to rounding and number of digits given for the MoS, this should be based on

the precision of the underlying data. The biological variability of toxicity data in vivo

generally is > 10%. The indication of more than decimal digits in the final MoS is therefore

not recommended.

3-5.2 THE THRESHOLD OF TOXICOLOGICAL CONCERN (TTC)

3-5.2.1 GENERAL CONCEPT OF TTC IN RISK ASSESSMENT

The use of the TTC approach as a risk assessment tool for cosmetics and consumer products

has been evaluated by the SCCS/SCHER/SCENHIR (SCCP/1171/08) as it is a pragmatic tool

that is based on the principle of establishing human exposure threshold values for all

chemicals below which there is a very low probability of an appreciable risk of systemic

adverse effects to human health.

Use of the TTC concept for chemicals with specific data requirements for their regulatory

approval under a specific European regulation is currently not acceptable as an alternative to

a chemical-specific evaluation.

Nevertheless, the TTC concept has been acknowledged to be a science-based prioritisation

and risk assessment tool by different organisations such as WHO IPCS, EFSA, SCCS, SCHER,

Health Canada (Joint FAO/WHO Expert Committee on Food Additives, 1996; SCCS, SCHER,

2012; EFSA, 2016a & 2019a; SCCS NoG 2016; Health Canada, 2016).

EFSA (EFSA, 2012 & 2019a) concluded that the TTC approach should not be used for the

following (categories of) chemicals: high potency carcinogens (i.e. aflatoxin-like, azoxy- or

N-nitroso-compounds, benzidines and also hydrazines); inorganic chemicals; metals and

organometallics; proteins; steroids; chemicals that are known or predicted to bioaccumulate;

nanomaterials; radioactive chemicals and mixtures of chemicals containing unknown chemical

structures.

_____________________________________________________________________________________________

So far, this approach has been used in a regulatory context for food contact material migrants,

food flavourings, fragrances, genotoxic constituents in herbal preparations and for pesticide

metabolites in groundwater.

The TTC approach aims to screen and prioritise chemical compounds for which the chemical

structure and exposure data are known, but for which no or limited toxicity data is available,

using an algorithm developed by Cramer (Cramer, 1978) where the substances, depending

upon their chemical structure, are grouped into three structural classes (low, medium, high

safety concern) in comparison with the toxicity data from available databases.

A database containing carcinogenicity data from animal studies for more than 3500

carcinogenicity experiments (Carcinogen Potency Database) (Gold et al., 1984) and a

database containing 613 chemicals based on toxicity other than carcinogenicity (Munro

database) (Munro et al., 1996) were available when the TTC approach was developed. Both

are based on systemic effects after oral exposure.

As with any risk assessment tool, application of the TTC approach requires a high level of

confidence in: 1) the quality and completeness of the databases; 2) the reliability of the

exposure data for the intended uses of the compound under study; and 3) the

appropriateness of any extrapolations.

3-5.2.2 TTC APPROACH FOR HUMAN HEALTH RISK ASSESSMENT OF CHEMICAL SUBSTANCES AND

COSMETIC SUBSTANCES

a) Systemic toxicity

The Scientific Committees (SCs) consider the TTC approach, in principle, scientifically

acceptable for human health risk assessment of systemic toxic effects caused by chemicals

present at very low levels. The application of the TTC should, however, be done on a case-

by-case basis and requires expert judgement. The TTC approach is also not applicable for a

number of chemical classes, which are indicated in detail in SCCP/1171/08 (adopted in 2012).

Practical application of the TTC approach to chemicals with no genotoxicity alert is usually

done by analysing the chemical structure and using Cramer classification as an indicator of

systemic toxicity. A small number of misclassifications of compounds when using the Cramer

decision tree in its present form have been revealed. Misclassification may also result in a

classification to a higher toxicity class. (Bhatia et al., 2015; Yang et al., 2017) and hence still

be conservative for safety evaluation.

The SCs concluded that the TTC value of Cramer Class II is not supported by the available

databases and these substances should be treated as Class III substances. The SCs also

accepted in principle the division of substances into Cramer Classes I or III (EFSA, 2016a).

When assigning a chemical to the lowest toxicity Class I, 1800 μg/person/d

corresponding to 30 μg/kg bw/d the classification should be carefully considered and

justified. If classification in Class I cannot be justified, the SCs recommended a general default

value equivalent to Class III compounds, being 90 μg/person/d, corresponding to

1.5 μg/kg bw/d for substances without genotoxicity alerts.

All the scientific information available today should be used to define the various toxicity

classes before expanding their number, i.e. the classification scheme should be modified

based on up-to-date toxicological knowledge (Boobis et al., 2017).

The SCCS agreed that, the default value of 0.15 μg/person/d, corresponding to

0.0025 µg/kg bw/d can be used for chemicals with genotoxicity alerts and hence

possible DNA reactive carcinogens but recommends its scientific basis to be strengthened.

This could be achieved by e.g., extending the database, analysing all available carcinogenicity

_____________________________________________________________________________________________

studies, using allometric adjustment factors and/or using the BMD

or BMD

as PoD for linear

extrapolation.

Usually, TTC values are expressed as an amount per person per day. In order to be applicable

to the entire population, including all age groups, it is advised to express TTC values in an

amount per kg body weight per day and give special consideration to infants under the age

of 6 months because of the potentially immature metabolism for some chemicals structures,

in particular when the estimated exposure is close to tolerable exposures defined by the TTC

values.

In the EU SEURAT-1 project COSMOS, work has been done on the TTC substances with non-

genotoxic alerts that are used for cosmetic purposes. The COSMOS TTC dataset, which was

quality controlled, contained 552 chemicals (495 cosmetic ingredients) with 219, 40, and 293

chemicals in Cramer Classes I, II, and III, respectively, to expand the chemical space and to

provide more robust thresholds for cosmetic-related chemicals. A TTC of 7.9 µg/kg bw/d was

suggested for Cramer Class III (which is 5-fold higher than the corresponding TTC value

derived by Munro et al., 1996). Cramer Class II was insufficient for derivation of a robust TTC

value. For Cramer Class I, a moderately increased TTC of 42 µg/kg bw/d was proposed. When

considering the COSMOS-plus-Munro i.e. “federated” dataset, values of 2.3 µg/kg bw/d

and 46 µg/kg bw/d were derived for Cramer Class III and I, respectively (Yang et

al., 2017). Although the TTC values are based on general toxicity data, it seems that datasets

specific for reproductive-developmental endpoints (Laufersweiler et al., 2012; van

Ravenzwaay et al., 2017) are adequately covered (Rogiers et al., 2020). Furthermore, work

of Patel et al. (2020) whereby 238, 76 and 162 fragrance chemicals in Cramer class I, II and

III of the RIFM TTC-database were integrated in the federated dataset, resulted in TTC values

for Cramer class I, II and III of 49.1, 12.7 and 2.9 µg/kg bw/ day, respectively. The different

values reported are taken up in Table 9 (PoDs used for derivation of TTCs are taken up in

APPENDIX 13).

It is important to note that an appropriate exposure assessment is essential for the

application of the TTC approach.

TTC thresholds are external dose-based values referring to oral systemic toxicity. For

cosmetics, the main exposure route is dermal. In the proposal from Kroes et al. (2007), an

external exposure value was converted to an internal exposure value by use of an adjustment

factor for percutaneous absorption. The latter value was then compared to the TTC value as

if the TTC value is also an internal exposure value. This is the case under the assumption of

100% oral bioavailability, which in many cases is an overestimation. For proper route-to-

route extrapolation, the NOAELs from the Munro database need to be corrected for oral

absorption. It should, however, be mentioned that in only few cases quantitative information

on absorption after oral administration is available.

_____________________________________________________________________________________________

Table 9: Overview of Threshold of toxicological concern (TTC) values (μg/kg bw/day).

Cramer

class

SCCP/1171/08

(Munro et al.

1996)

Cosmos-TTC

(SEURAT-1;

European

commission,

2009)

Cosmos/Munro/

Federated DB

(Yang et al., 2017)

RIFM/Munro/Cosm

os/Federated DB

(Patel et al., 2020)

Genotoxic

compounds

0.0025

49.1

II*

Not supported**

12.7

III*

1.5

7.9

2.3

2.9

*Non-genotoxic compounds

**Chemicals of Cramer class II should be treated as Class III substances.

Values in bold are those currently recommended by the SCCS for use for cosmetics-related

substances.

For botanical extracts, Kawamoto et al. (2019) reported that the Cramer class III TTC value

of 90 µg/person/d might be adequately conservative. For potentially genotoxic substances a

TTC value of 10 µg of plant material on a dry weight basis/person per day has been proposed

(Mahony et al., 2020). These values are not taken up in the Table 9 as plant materials are

composed of mixtures.

TTC thresholds are external dose-based values referring to oral systemic toxicity. For

cosmetics, the main exposure route is dermal. In the proposal from Kroes et al. (2007), an

external exposure value was converted to an internal exposure value by use of an adjustment

factor for percutaneous absorption. The latter value was then compared to the TTC value as

if the TTC value was also an internal exposure value. This is the case under the assumption

of 100% oral bioavailability, which in many cases is an overestimation. For proper route-to-

route extrapolation, the NOAELs from the Munro database need to be corrected for oral

absorption. It should, however, be mentioned that only in a few cases is quantitative

information on absorption after oral administration available.

The SCCS considers that at present the thresholds proposed by the ‘federated Yang et

al. (2017)’ data set of 2.3 µg/kg bw/d and 46 µg/kg bw/d for Cramer classes III

and I respectively, are appropriate for use in relation to cosmetics-related

substances.

b) Inhalation toxicity

For inhalation exposure TTC, only limited information is available (Carthew et al., 2009;

Escher et al., 2010; Schüürmann et al., 2016). Compared to the existing oral database, the

pool of available repeated dose inhalation exposure studies is scarce (about 400 rodent

studies and even fewer with accompanying local respiratory effects observations) (RIFM

database). The development of inhalation TTC is not yet mature enough to be considered as

a valid risk assessment tool.

3-5.2.3 ITTC APPROACH

For cosmetic ingredients any risk assessment as well as the TTC approach should be based

on internal doses (Partosch et al., 2014). Therefore, when the TTC approach is applied for

cosmetic ingredients, an adjusted internal TTC value has to be defined considering both

dermal and oral absorption. As such, several attempts have been made to arrive to an

_____________________________________________________________________________________________

internal TTC (iTTC) by adjusting the external NOAEL values of substances by in silico

estimates of oral bioavailabilty (Partosch et al., 2015, Reilly et al., 2019). However, the

estimates were still based on external dose and not an internal exposure metric such as

plasma concentration.

Within the framework of a multi-stakeholder project, further work is currently ongoing

towards the development of a set of robust iTTC values that could be utilised in human safety

assessment. It is, however, clear that developing an iTTC database is complex and more

research is required beyond current attempts where NOAELs were only adjusted for by

applying in silico tools (Ellison et al., 2019; Rogiers et al., 2020). While work is ongoing to

develop robust iTTC thresholds, an interim conservative iTTC of 1 μM plasma concentration

for chemicals in consumer products has been proposed, which is supported by the published

experience of the pharmaceutical industry, a literature review of non-drug chemical/receptor

interactions, and analysis of ToxCast™ data. This is, however, with the additional exclusion

to the original TTC exclusion criteria of the estrogen and androgen receptors as targets of

concern for low dose exposures.

Efforts are still ongoing to further extend/ refine the TTC framework for inhalation TTC and

internal TTC. From the point of view of NAMs, it is clear that the TTC and/or iTTC concepts

will be of great value in the future.

3-6 SPECIAL CONSIDERATION FOR CERTAIN COSMETIC INGREDIENTS

3-6.1 MULTI-CONSTITUENT NATURAL INGREDIENTS

Many cosmetic ingredients can be mixtures of multiple substances of natural origin, e.g.

essential oils and fragrances; they often can considerably vary in their composition depending

on their geographical origin, conditions of harvest, storage, further technical processing etc.

In such cases, the cosmetic ingredient should contain the following information:

− qualitative identification and semi-quantitative concentrations of the substances in the

mixture (e.g. <5%) using the preferred terminology as indicated in Section II of the

Inventory of Cosmetic Ingredients and the INCI/CIN name if available;

− for mixtures of variable composition, an indication of the range and the maximum

levels of components which may be present in the mixture, taking into account batch

to batch variation;

− a clear indication of the cosmetic product category in which the mixture may be used

and at what maximum concentration.

− Case by case, in the final safety evaluation, reference should be made to the semi-

quantitative composition of the multi-constituent ingredient and the toxic potential of

components should be considered.

− Specific labelling to reduce the incidence of contact-allergic reactions in fragrance-

sensitive consumers has been foreseen by the inclusion of 26 potentially sensitising

fragrance substances in Commission Regulation (EU) 2019/831 amended Annex III to

Regulation (EC) No 1223/2009.

More specifically, the presence of these substances must be indicated in the list of substances

on the label when their concentrations in the final product exceed 0.001% in leave-on

products or 0.01% in rinse-off products (2003/15/EC).

The SCCS has adopted an Opinion on fragrance allergens in cosmetic products which enlarges

the list of fragrance allergens considered relevant for consumers and which makes it possible

to derive a general threshold for substances with a higher number of recorded cases

(SCCS/1459/11).

_____________________________________________________________________________________________

3-6.2 IDENTIFICATION OF MINERAL, ANIMAL, BOTANICAL AND BIOTECHNOLOGICAL INGREDIENTS

IN A COSMETIC PRODUCT

The nature and preparation of some substances may affect the type and amount of data

necessary for their identification. The following points indicate the advised requirements for:

a) Complex substances of mineral origin

starting material

description of:

- the preparation process: physical processing, chemical modifications, possible

purification,

- characteristic elements of the composition: characteristic components, known toxic

components (%).

physical and chemical specifications

microbiological quality

preservatives and/or other additives added.

b) Complex substances of animal origin

When animal-derived cosmetic substances are used, this should be clearly mentioned (see

3.6.3)

species (bovine, ovine, crustacean, …)

organs, tissues, biological liquids (placenta, serum, cartilage, ...)

country of origin

description of:

- the preparation process: conditions of extraction (solvent, pH, temperature, …); type

of hydrolysis (acidic, enzymatic, …); other chemical modifications; possible purification;

- commercial form: powder, solution, suspension, freeze-dried, …

- characteristic elements of the composition: characteristic amino acids, total nitrogen,

proteins, polysaccharides, molecular mass, …

physical and chemical specifications

microbiological quality including relevant viral contamination

additional external contamination

preservatives and/or other additives added.

c) Complex substances of botanical origin

common or usual names of the plant, alga or macroscopic fungus

name of variety, species, genus, and family

in case more than one variety of source of a given species is used, each should be specified

organoleptic, macroscopic and microscopic evaluation

_____________________________________________________________________________________________

morphological and anatomical description (including gender, if applicable) and a

photograph of the plant or plant part, alga, or macroscopic fungus used

natural habitat and geographical distribution of the plant, alga, or macroscopic fungus

current sources of the plant, alga, or macroscopic fungus, including its geographical

location and whether it is cultivated or harvested from the wild

description of:

- preparation process: collection, washing, drying, extraction, distillation, destructive

distillation, possible purification, preservation procedures, …;

- handling, transportation, storage;

- commercial form: powder, solution, suspension, …;

- characteristic elements of the composition: identification of characteristic components,

known toxic components (%);

physical and chemical specifications

microbiological quality including relevant fungi

additional external contamination

preservatives and/or other additives added.

d) Complex substances derived from biotechnology

For special biotechnologically derived substances, where a modified microorganism or a

potential toxic substance has not been fully removed, specific data must be available, which

can comprise:

description of organisms involved: donor organisms, recipient organisms, modified

microorganisms

host pathogenicity

toxicity, and when possible, identity of metabolites, toxins produced by the organisms

fate of viable organisms in the environment-survival-potential for transfer of

characteristics to e.g. natural bacteria

physical and chemical specifications

microbiological quality

additional external contamination

preservatives and/or other additives added.

3-6.3 ANIMAL-DERIVED COSMETIC SUBSTANCES

When animal derived cosmetic substances are used, this should be clearly mentioned.

Entry no. 419 in Commission Reg. (EU) 2019/831 amended Annex II of Reg. 1223/2009/EU

specifies several substances for which some concern exists for human health with respect to

Transmissible Spongiform Encephalopathy (TSE).

“419. Category 1 material and Category 2 material as defined in Articles 8 and 9, respectively

of Regulation (EC) No 1069/2009 of the European Parliament and of the Council of 21

October 2009 and substances derived therefrom

.”

OJ L 300, 14.11.2009, p. 1

_____________________________________________________________________________________________

As indicated, tallow derivatives of bovine origin are considered as an exception and are

accepted as cosmetic substances provided they undergo a number of specific treatments.

At present, there is no evidence that TSE may be transmitted by topical exposure.

Finally, taking into account EC Regulation No 1069/2009 laying down health rules concerning

animal by-products not intended for human consumption, the SCCP was of the opinion that

substances derived from category 1 (inter alia specific risk material) and category 2 (inter

alia 'fallen stock') material raise concern in terms of biological risk for human health and

therefore must not be present in cosmetic products (SCCP/0933/05). Category 3 material is

not intended for human consumption, but it may be used as cosmetic substance in accordance

with Regulation 1069/2009, Article 33.

Non-animal derived supplements for in vitro testing should be used wherever possible. The

chemically defined/serum-free cell culture media can be found in several in vitro test methods

for skin corrosion, skin irritation and eye irritation testing (OECD 431, 439 and 492) (van der

Valk et al., 2017).

3-6.4 SUN PROTECTION SUBSTANCES

For sunscreen lotion, an amount of 18.0 g/day is used in the MoS calculation. It is used

as a standard exposure value in the safety evaluation carried out by the SCCS but is not

meant as a recommended amount to be applied by the consumer (SCCNFP/0321/02).

To reach a comparable level as indicated by the Sun Protection Factor (SPF), sunscreen

products have to be applied in quantities similar to the ones used for SPF testing, i.e. 2

mg/cm

(total amount of approx. 36 grams) for the body of an average adult person

(2006/647/EC). The quantity of 2 mg/cm

, however, is the amount necessary to obtain

reproducible SPF results under laboratory conditions. It is higher than the amount usually

applied by consumers.

This observation has been reported frequently: when consumers use their own sun products

(lotions, alcoholic solutions, gels, creams, sprays,…) and apply the products on the whole

body surface, values for use of products between 0.5 - 1.3 mg/cm² have been found

(Stenberg et al., 1985; Bech-Thomsen et al., 1993; Diffey, 1996; Gottlieb et al., 1997; Autier

et al., 2001 and 2007). The values seem to depend on the study protocol used, the location

on the body measured and several other factors. More recent publications still come to

comparable values in the range of 0.39-1 mg/cm² (Danish Protection Agency No. 151, 2016,

Ficheux et al., 2016a, Gomez-Berrada et al., 2017). When the product is applied only to the

face, then the amount applied might be higher than 2 mg/cm

(Gomez-Berrada et al., 2017).

The amount used by the SCCS in safety calculations reflects actual consumer use and takes

the whole body area (17500 cm

) into account. The average exposed skin area of sunscreen

users according to the recent report of the Danish authorities is 14,700 cm

The use of 18g/d sunscreen corresponds with the values reported by Biesterbos et al.

(Biesterbos et al., 2013), who found a mean use amount of 9.2 g/application, derived on the

basis of pictures. If two applications are considered, this is about 18 g/d. Unpublished data

by von Goetz (von Goetz, 2018) from a small-scale pilot study with weighing also provided a

mean of 9 g for whole-body application (5 applications by 2 persons).

If a sun protection substance is applied in a sprayable product that may give consumer lung

exposure by inhalation, other considerations should be taken into account (see 3-3.4.1.3).

For lipcare products, 100% absorption of the substance should be considered for safety

assessment.

3-6.5 ENDOCRINE ACTIVE SUBSTANCES (EAS)

3-6.5.1 DEFINITIONS

Some natural and synthetic chemical substances can interact, interfere or disrupt the function

of the endocrine system that regulates various metabolic and developmental functions in the

_____________________________________________________________________________________________

body (WHO/IPCS, 2002; UNEP/WHO, 2012). The endocrine system comprises a complex

array of signalling and feedback mechanisms, the disruption of which has been linked to

various adverse health effects, such as reproductive effects, metabolic disorders, cognitive

deficits and cancers. However, the endocrine system also involves numerous cycles and

feedback loop mechanisms and adaptive responses that together regulate the secretion of

hormones and maintain homeostasis. A substance interfering with the endocrine system may

affect hormone secretion or other cellular factors, but it is possible that such perturbations

remain within the homeostatic or metabolic detoxification capacity and therefore do not result

in adverse effects in the intact organism. Some effects linked to endocrine disruption have

also been shown to have critical window(s) of susceptibility, e.g. increased susceptibility of

an organism within a certain developmental period.

-The definition of Endocrine Disrupters (EDs) endorsed at the European level

is the same as

proposed by WHO/IPCS (WHO/IPCS, 2002) and is as follows: “An endocrine disruptor is an

exogenous substance or mixture that alters function(s) of the endocrine system and

consequently causes adverse health effects in an intact organism, or its progeny, or

(sub)populations”.

-The OECD's revised conceptual framework (OECD TG 150) also has a prerequisite to identify

the adverse effect in an intact organism for regarding a substance an endocrine disruptor.

Thus, whilst a chemical may be regarded an EAS on the basis of activity/interaction towards

one or more components of the endocrine system (e.g., a hormone receptor), it can only be

regarded as an ED if there is evidence for a biologically-plausible causal relationship between

the endocrine perturbation/activity and the adverse effect(s) in an intact organism.

-The joint EFSA/ECHA/JRC draft guidance (EFSA and ECHA, JRC, 2018) has defined endocrine

activity as 'Interaction with the endocrine system which can potentially result in an effect on

the endocrine system, target organs and tissues'

3-6.5.2 IDENTIFICATION OF EDS AND REGULATORY CONSEQUENCES

A number of chemicals have been identified, or are suspected, as EDs. However, "only a small

fraction of these chemicals has been investigated in tests capable of identifying overt

endocrine effects in intact organisms" (WHO-UNEP report, 2012).

Under REACH, EDs can be identified as Substances of Very High Concern (SVHC) alongside

chemicals known to cause cancer, mutations and toxicity to reproduction. There are several

substances identified as SVHC for their endocrine disrupting properties in the Candidate List

of SVHC for authorisation (https://echa.europa.eu/candidate-list-table).

Amongst other actions, the Commission launched the Fitness Check: https://eur-

lex.europa.eu/legal-content/EN/TXT/?qid=1553617067256&uri=CELEX:52018DC0734 and

regulated ED substances in specific areas, including chemicals (Regulation EC 1907/2006),

Pesticides (Regulation EC 1107/2009), Biocides (Regulation EU 528/2012), Water quality

(Water Framework Directive 2000/60/EC).

3-6.5.3 STEPWISE APPROACH FOR COSMETICS AND THEIR INGREDIENTS

For cosmetics, the Commission adopted a review of the Cosmetics Regulation regarding

substances with endocrine disrupting properties

. It was concluded that adequate tools are

available to regulate the use of cosmetic substances that present a potential risk for human

Commission Regulation (EU) 2018/605 of 19 April 2018 amending Annex II to Regulation (EC) No 1107/2009 by

setting out scientific criteria for the determination of endocrine disrupting properties. OJ L 101, 20.4.2018, p. 33–

36.

_____________________________________________________________________________________________

health, including when displaying ED properties. For environmental concerns, application of

the REACH Regulation’ is considered.

The SCCS is following this process closely and is actively engaged in the safety assessment

of potential ED substances used in cosmetics.

Due to the animal testing ban under the Cosmetics Regulation, it is now out of scope to test

cosmetic ingredients in vivo for endocrine disruption. Cosmetic ingredients therefore can be

assessed for endocrine activity in a stepwise approach using data generated outside the

cosmetic field or for a new cosmetic ingredient, using NAMs (in silico models, read across, in

vitro assays, other mechanistic techniques such as 'omics').

Among the various endocrine modalities, Estrogen (E), Androgen (A), Thyroid (T) and

Steroidogenic (S) - (EATS) pathways are the best characterised, whereas retinoid signalling

and hypothalamo-pituitary-thyroid axis are poorly investigated (Kortenkamp et al., 2011;

UNEP/WHO, 2012).

The OECD 150 guideline provides tools on how to assess endocrine properties of a substance.

The general approach taken by this GD is primarily to consider the possible results that might

be obtained from each endocrine disruption-responsive assay and to provide guidance about

how these results might be interpreted in light of data that may or may not already be

available from other in vitro or in vivo assays. This should include all available data such as

publications in the peer-reviewed literature as well as TGs. In order to inform this

interpretation, background data on the assays addressed, non-testing approaches and other

considerations relevant to the assays are discussed. These include cross-species

extrapolations, read-across and multiple Modes of Action (MoA). The nature, quantity and

quality of the existing and new data in each of the scenarios for the endocrine disruption-

responsive assays should be evaluated systematically in a WoE approach. There is generally

no single “right” answer. Use of other technologies (e.g. “omics” data) may help in

understanding the link between endocrine-related mechanisms and a WoE approach. This GD

should therefore be used flexibly in light of local regulatory needs. The key questions

addressed concern likely mechanisms of endocrine action and any resulting apical effects that

can be attributed to such action. In Table 10 the conceptual framework for testing and

assessment of EDs as provided in OECD guideline 150 is shown.

_____________________________________________________________________________________________

Table 10: OECD conceptual framework for testing and assessment of EDs

QSARS = Quantitative Structure Activity Relationship;

For a new cosmetic ingredient, due to the animal testing ban, characterisation will, however,

be limited to the study of endocrine activity at level 1 (existing data and using in vivo data

if they have been generated before the animal ban or for another regulatory purpose than

cosmetics) and level 2 (in vitro assays) of the OECD's revised Conceptual Framework as

described below.

● Lines of evidence level-1 (existing data and non-test information):

The first level of evidence for endocrine activity of a substance may be provided by: physical

and chemical properties (e.g., MW, reactivity, volatility, biodegradability), all available

(eco)toxicological data from standardised or non-standardised tests, read-across, chemical

categories, QSARs and other in silico predictions, and ADME model predictions for a new

compound intended for use in a cosmetic product, the use of in silico models and read-across

tools, together with physicochemical data.

A number of in silico models and tools are available for the estimation of a substance's

potential for binding with hormone receptors, such as the Estrogen Receptor (ER), the

Androgen Receptor (AR), and the Pregnane X Receptor (PXR). These include commercial

programmess such as ADMET Predictor™ and MetaDrug™, as well as publicly available tools

such as VEGA and Online Chemical Modeling Environment (OCHEM). Another open source

docking tool, Endocrine Disruptome, is also available for virtual screening of EDs (see EFSA

and ECHA, JRC, 2018).

In addition, databases are available that provide some information on endocrine properties

of chemical substances

. These may be subject to some criticism (e.g., inaccurate

information, some entries not enough documented). Endocrine Disruptor Screening Program

(EDSP) Tier 1 screening assay results and the dataset from Collaborative Estrogen Receptor

Endocrine active substances information system (EASIS) (EC JRC); ToxCast (US EPA); ToxCast ER prediction

model (US EPA); SIN List (International chemical secretariat); The endocrine disruption exchange (TEDX);

Endocrine disruptor screening program, EDSP21 (US EPA); Endocrine disruptor knowledge base, EDKB database

(US FDA); Estrogenic activity database, EADB (US FDA); Toxicology data network (Toxnet); Developmental and

Reproductive Toxicology database (DART); NURSA (nuclear receptor signalling atlas); OECD (Q)SAR toolbox

(OECD, ECHA); AOP knowledge base (OECD); ToxRefDB (US EPA); eChem portal (OECD); COSMOS database -

cosmetic ingredients; Danish (Q)SAR Database; (Q)SAR Data Bank

_____________________________________________________________________________________________

Activity Prediction Project (CERAPP) are also reported in Mansouri et al., 2016. These

databases may also enable read-across for endocrine activity and provide a basis for further

development of structure-activity based predictive models. Some of these databases also

contain in vivo experimental data.

Amongst the available in silico tools, the OECD QSAR Toolbox offers a major software platform

that incorporates several databases comprising chemical data, experimental

(eco)toxicological data, and estimated values from QSAR tools, together with incorporated

QSAR modelling tools and Expert Systems. For example, it contains:

- The OASIS Estrogen Binding Database, consisting of diverse compounds with relative

Endocrine Receptor Binding Assay (ERBA) data. The Toolbox allows in silico screening of

a compound’s endocrine activity through Danish EPA's Relative ERBA (Q)SAR, which is

based on ER binding in vitro.

- QSAR models, including MultiCASE ERBA QSAR, which is based on a hierarchical statistical

analysis of a training set composed of ER binding data on a variety of chemical structures

that are inactive, weak, or powerful ER binders.

- Structural-alert based ER-binding profiler to classify chemicals as non-binders or binders

(weak, moderate, strong and very strong binders) depending on their MW and structural

characteristics.

- Structural-alert based expert systems, such as the US EPA's rtnER expert system based

on binding to the rainbow trout estrogen receptor.

The OECD QSAR Toolbox also provides a major platform for read-across between chemicals

that share structural and/or functional similarities, using a substantial set of high quality

databases. If compounds in the database are identified with the required structural and alert

profile similarities to the target compound, they may be used as read-across candidates for

the prediction of the ER binding of the target compound.

Other in silico systems based on molecular docking tools and 3D-(Q)SAR models are also

available that allow virtual screening of chemical substances for affinity/binding with hormone

receptors (Jacobs, 2004; Vedani et al., 2012; Galli, 2014). The identification of

affinity/binding to a hormone receptor by virtual screening, however, needs to be seen in the

context of the scoring function used for each target, because a universally applicable scoring

function is not yet available (Vuorinen et al., 2013). Also, whilst in silico models can reliably

predict simple endpoints, such as the binding free energy toward the receptor binding, they

have a limitation for the prediction of more complex endocrine related in vivo endpoints, such

as reproductive and developmental toxicity.

The available experimental data are still too scarce to allow comparison between the success

rates of the results from different in silico methods (Vuorinen et al., 2013). The topic has

been recently reviewed by Schneider et al. (2019), who highlighted that whilst in silico

prediction approaches provide first stage indication of ED properties, further modeling of

intermolecular interactions and cellular behavior is also essential to understand the potential

effects on the endocrine system.

● Lines of evidence level-2 (in vitro assays providing data about selected

endocrine mechanism(s)/ pathways(s) (mammalian and non-mammalian

methods).

The currently available in vitro methods include estrogen, androgen, or steroidogenic receptor

binding assays, whilst methods relevant to thyroid hormone are not sufficiently sensitive to

completely exclude effects due to disruption of thyroid-related functions. A validation study

on 17 methods for the detection of thyroid disruptors was launched by EURL ECVAM (JRC

2017). The available in vitro methods are listed below:

_____________________________________________________________________________________________

- Estrogen (OECD TG 493) or androgen receptor binding affinity (US EPA TG OPPTS

890.1150) (OPPTS stands for Test guidelines for pesticides and toxic substances).

- Estrogen receptor transactivation (OECD TG 455),

- Yeast estrogen screen (ISO 19040-1,2&3)

- Androgen receptor transcriptional activation (OECD TG 458)

- Steroidogenesis in vitro (OECD TG 456)

- Aromatase Assay (US EPA TG OPPT 890.1200)

- Thyroid disruption assays (e.g., thyroperoxidase inhibition, transthyretin binding)

- Retinoid receptor transactivation assays

- Other hormone receptors assays as appropriate

- High-throughput screens (See OECD GD No. 211 describing Non-Guideline In vitro Test

Methods: OECD 2014c)

Whilst the results from Levels 1 and 2 approaches can be indicative of endocrine activity of a

cosmetic ingredient, they will not definitively inform whether the substance will cause adverse

effect(s) in the intact organism to be regarded an ED. In view of this limitation, it is important

that all the evidence from physicochemical properties, available literature, in silico models,

read-across, in vitro assays, and other techniques (such as “-omics”) is integrated in a

systematic manner to generate sufficient WoE to exclude the potential toxicity of a cosmetic

ingredient through the endocrine related effects. The integration of in silico methods and

computational systems biology has been proposed as a means to more critically assess the

endocrine activity of chemical substances (Ruiz et al., 2017). Some key characteristics of EDs

have also been proposed following an expert consensus statement as a basis for hazard

identification (Merill et al., 2020).

3-6.5.4 COSMETIC INGREDIENTS SUSPECTED TO HAVE ED PROPERTIES

As yet there is no harmonised approach towards health risk assessment procedures for EDs

within the different regulatory frameworks in the EU. The SCCS has issued a memorandum

(SCCS/1544/14) to clarify its position on substances with potential ED properties when used

as cosmetic ingredients. In the context of the animal testing ban, it is not possible for the

SCCS to fulfill the criteria as laid out under the OECD Conceptual Framework for the

identification of EDs for cosmetic ingredients in the context of the animal testing ban.

In the view of the SCCS, these substances should be treated like other substances of concern

for human health and therefore be subject to risk assessment and not only hazard

assessment.

This is in agreement with the past and current evaluations by the SCCS in regard to the safety

assessment of cosmetic ingredients with suspected ED properties e.g., parabens

(SCCP/1017/06, SCCP/1183/08, SCCS/1348/10, SCCS/1446/11, SCCS/1514/13), triclosan

(SCCP/1192/08, SCCS/1414/11), homosalate (SCCP/1086/07), benzophenones, 4-

methylbenzylidene camphor and 3-benzylidene camphor (SCCNFP/0483/01, SCCP/1183/08,

SCCS/1513/13), melatonin (SCCS/1315/10), resorcinol (SCCS/1270/09), cyclomethicone

(SCCS/1241/10), decamethylcyclopentasiloxane (cyclopentasiloxane) (SCCS/1549/15).

Ingredients with potential endocrine disrupting properties used in cosmetic products are taken

up in a list of 28 compounds to be considered by the SCCS for safety evaluation. 14

substances of this list are considered high priority and are currently being assessed by the

SCCS. These are benzophenone-3, kojic acid, 4-methylbenzylidene camphor, propylparaben,

triclosan, resorcinol, octocrylene, triclocarban, butylated hydroxytoluene (BHT),

benzophenone, homosalate, benzyl salicylate, genistein and daidzein,

Another way forward could be to demonstrate what could be considered as biologically

irrelevant exposure. For instance, in the case of melatonin, topical application (in real use

conditions) did not perturb endogenous hormone levels in humans due to low systemic

exposure (SCCS/1315/10). Toxicokinetic studies and PBPK modelling could help to bridge the

_____________________________________________________________________________________________

gap between in vivo and in vitro evidence by providing data on (internal) exposure in relation

to concentrations that were found to be active in in vitro assays (Coecke et al., 2013; Bessems

et al., 2014).

It also needs to be highlighted that the SCCS only assesses cosmetic ingredients in relation

to safety of consumers' health, and as such they are not assessed for effects on the

environment. Data generated on the environmental effects may, however, be also useful to

support EA/ED mode of action but not their potency. For example, some ecotox tests may be

informative for the assessment of endocrine activity of a compound in humans or thyroid

effects (e.g. Xenopus Eleutheroembryonic Thyroid Assay (XETA) (OECD 248), Amphibian

Metamorphosis Assay (AMA) (OECD Test N° 231), Larval Amphibian Growth and Development

Assay (LAGDA) (OECD Test N° 241).

A recent review has indicated a high degree of confidence in the conservation of the HPG-

axis between fish and mammals, and the HPT-axis between amphibians and mammals

(McArdle et al., 2020).

An ongoing EU project ERGO (https://ergo-project.eu/) is looking into the scientific basis that

could bridge the current divide between human health and the environment in terms of non-

mammalian testing for the identification of EDs (with a focus on the thyroid system) for the

chemicals that affect endocrine axes across vertebrate classes.

3-6.6 CMR SUBSTANCES

Based on their inherent properties, hazardous chemicals are classified accordingly on a world-

wide (Globally Harmonised System) and European level (Regulation 1272/2008). Special

attention is given to substances that are carcinogenic, germ and somatic cell mutagenic or

toxic for reproduction for which three hazard classes exist according to these frameworks,

i.e. Category 1A, 1B and 2. Cat 1A means that the substance is known to have the respective

potential in humans, Cat 1B means that the substance is presumed to have the respective

potential in humans, and Cat. 2 means that the substance is suspected to have the respective

potential in humans.

CMR 1A, 1B and 2 substances are prohibited for use in cosmetics, unless the specific criteria

set in Cosmetics Regulation (EC) No 1223/2009 are fulfilled, whereby criteria are stronger

for CMR 1A and 1B substances compared to CMR 2 substances

CMR 2 substances may be used in cosmetics where they have been evaluated by the SCCS

and found safe. These substances could be allowed to be used as cosmetic substances within

Europe under specific conditions. Examples for CMR2 substances include trisodium

nitriloacetate (SCCS/1391/10), trimethylbenzoyldiphenylphosphine oxide (TPO)

(SCCS/1528/14) polyaminopropyl biguanide (PHMB) (SCCS/1581/16), lysmeral

(SCCS/1591/17), salicylic acid (SCCS/1601/18), pigmentary TiO

(SCCS/1617/20).

CMR 1A or 1B substances may be used in cosmetics exceptionally where (1) they comply

with the European food safety requirements

, (2) they cannot be replaced by suitable

alternatives, (3) the application is made for a particular use of the product category with a

known exposure and (4) the substances were evaluated and found safe by the SCCS for use

in cosmetic products, in particular in view of exposure to these products and taking into

consideration the overall exposure from other sources, taking particular account of vulnerable

population subgroups (2009/1223/EC). Examples for CMR 1B substances include boron

compounds (SCCS/1523/13), formaldehyde in nail hardener (SCCS/1538/14) and zinc

pyrithione (SCCS/1614/19).

A guidance document has been developed by the EU Commission with the aim of enabling a

harmonised approach to the development and use of aggregate exposure estimates in

assessing the safe use of CMR substances as cosmetic ingredients (see Appendix 5).

Regulation (EC) No. 178/2002

_____________________________________________________________________________________________

However, as clarification and as agreed by the Commission, whereas the applicant is

responsible for providing the exposure data on CMR substances, the procedure described in

No. 16-19, 21 and 22 of the Guidance, is only foreseen in case that the applicant for any

reason cannot obtain the data from the owner of the data required.

3-6.7 LIFETIME CANCER RISK (LCR)

In the safety assessment of carcinogenic substances, an appropriate dose descriptor, BMDL10

or T25, should be identified, whenever sufficient information is available (ECHA, 2019; EFSA,

2019b; COC, 2020). The SCCS recommends that, where possible, the BMD approach should

be used for deriving a POD, as a starting point for human health risk assessment, including

carcinogenicity by a genotoxic or non-genotoxic mode of action. This view is also supported

by other bodies including the EFSA and the Committee on Carcinogenicity of Chemicals in

Food, Consumer Products and the Environment (COC). In the absence of dose-response data

allowing for the application of the BMD approach, the T25 is a simplified method to estimate

the carcinogenic potency of a given substance.

The T25 (expressed as mg/kg bw/d) is defined as the dose which leads to the development

of tumours at a specific tissue site in 25% of the animals after correction for spontaneous

incidence and within the standard lifetime of the species (Dybing et al., 1997). The

determination of BMDL10 (expressed as mg/kg bw/d) uses mathematical curve fitting

techniques to calculate the lower 95% confidence level at a 10% benchmark response. Both

BMDL10 and T25 can be used as starting points to determine an additional LCR or to calculate

a MoE, which represents the ratio between a dose descriptor and the estimated human

exposure dose. Basic steps in LCR calculations based on T25 are provided in Appendix 12.

Some countries and international organisations have considered that the LCR in the general

population of less than 10

-5

is considered tolerable (SCCS/1486/12). Under REACH, the

"indicative tolerable cancer risk level" for the general population is 10

-6

(ECHA 2012a). It

should be noted that the tolerable LCR is a risk management issue and outside the scope of

the mandate of the SCCS.

3-6.8 NANOMATERIALS

3-6.8.1 DEFINITION OF NANOMATERIAL

Regulation (EC) No 1223/2009 specifically covers the use of nanomaterials in cosmetic

products. The Regulation provides a definition of nanomaterial, as well as a mechanism for

notification, labelling, and safety evaluation of cosmetic products containing nanomaterials.

Under Article 2 (1) (k), “nanomaterial” means an insoluble or bio-persistent and intentionally

manufactured material with one or more external dimensions, or an internal structure, on the

scale from 1 to 100 nm”.

The Regulation therefore covers mainly those nanomaterials that are intentionally produced

and are insoluble/poorly-soluble or biopersistent (e.g., metals, metal oxides, carbon

materials, etc.), and not those that are either completely soluble or degraded and are not

persistent in biological systems (e.g., liposomes, oil/water emulsions, etc.).

When dealing with the question of solubility, as provided in the current definition, it is

important to note that any nano-specific risk may change (even diminish) when a

nanomaterial is dissolved. But it is the time period during which the dissolution happens that

determines the considerations for risk assessment based on either particle risk or soluble

substance risk. Partial dissolution over a long period of time may lead to the mistaken claim

_____________________________________________________________________________________________

that the material is 'soluble', and therefore not a nanomaterial under the scope of the current

definition provided in the Cosmetic Regulation (EC) No 1223/2009.

3-6.8.2 POTENTIAL SAFETY ISSUES OF NANOMATERIALS

The use of nanomaterials in cosmetics is subject to a high level of protection of human health

under the EU Cosmetics Regulation. This is because nano forms of some substances may

differ from their conventional (bulk) forms in terms of physicochemical properties, biokinetic

behaviour, and/or biological effects. Any intended use of nanomaterials (other than

colourants, preservatives and UV filters and not otherwise restricted by the EU Cosmetics

Regulation) in cosmetic products must be notified to the Commission by the RP through the

Cosmetic Product Notification Portal (CPNP) at least six months prior to placing them on the

market, except if they were already on the market before 11 January 2013. In case of a

safety concern over a nanomaterial, the Commission shall request the SCCS for a scientific

Opinion on the safety of the nanomaterial for use in relevant categories of cosmetic products

in consideration of the reasonably foreseeable consumer exposure.

The SCCS was recently mandated by the Commission to provide scientific advice to facilitate

the identification of any safety concerns relating to the nanomaterials intended for use in

cosmetic products, so that they can be prioritised for safety assessment. The advice has

recently been published (SCCS/1618/2020), which provides the key scientific aspects of a

nanomaterial that should trigger consumer safety concerns, and therefore the need for further

evidence-based safety assessment.

Although there are currently no hard and fast rules for identifying the safety concerns for

nanomaterials, as a general principle, each of the following attributes should add a further

degree of safety concern. For example, where:

1. The nanomaterial has constituent particles that have sizes in the lower range of the

nanoscale.

2. The nanomaterial is insoluble, or only partially soluble.

3. The chemical nature of the nanomaterial suggests the potential for a toxicological hazard.

4. The nanomaterial has certain physical/morphological features (e.g. needle shape, rigid

long fibres) that point to the potential for harmful effects.

5. The nanomaterial has surface reactivity in terms of catalytic (including photocatalytic)

activity, potential for radical formation, or other surface properties (e.g. potential

allergenicity due to proteinaceous surface).

6. The nanomaterial has a different biokinetic behaviour than the conventional equivalent.

For example, a surface modification/coating (e.g. hydrophobic coatings, encapsulation)

has been applied to core nanoparticles to alter their ADME properties and as a result make

them more accessible systemically, compared to the neat nanoparticles and/or their

conventional chemical forms.

7. The nanomaterial is used as vehicle to carry other substances that have not been assessed

for safety as individual components, or together in the form of nano-scale entity.

8. There is a likelihood of systemic exposure of the consumer to nanoparticles through the

use of final products. The frequency of use, and/or the amounts of the relevant consumer

product are relatively high.

9. There is evidence for persistence/accumulation of nanoparticles in the body.

10. Nanoparticles have other distinctive properties not present in conventional form of the

same material, or have a new activity/function (e.g. a smart/functional nanomaterial).

_____________________________________________________________________________________________

11. The nanomaterial is so novel that it does not have a conventional comparator to allow

assessment of changes in properties, behaviour or effects.

12. The nanomaterial is used in a product that is inhalable (taken up by inhalation into

respiratory tract and lung), and the particles are respirable (can reach respiratory

epithelium i.e. alveoli).

13. The assessment of genotoxicity is performed inadequately, e.g. in vitro studies are without

information on stability of the test suspension, or evidence of cell exposure

(internalisation).

Whilst this section only provides a brief guidance on nanomaterials in cosmetics, the SCCS

has published a more detailed specific Guidance on Risk Assessment of Nanomaterials

(SCCS/1611/19), which is an update of a previous guidance published in 2012

(SCCS/1484/12), a Memorandum on the Relevance, Adequacy and Quality of the Data

Expected in Safety Dossiers on Nanomaterials (SCCS/1524/13, Revision of 27 March 2014),

and a checklist for the applicants submitting dossiers on nanomaterials as cosmetic

ingredients (SCCS/1588/17).

Safety assessors need to consult these documents to ensure that any testing to generate

evidence on the safety of nanomaterials is carried out with special considerations of the nano-

size related characteristics of the materials, and in compliance with the ban on animal testing

of cosmetic ingredients. In this regard, it is important to note that, as indicated in the

memorandum (SCCS/1524/13, Revision of 27 March 2014), the SCCS will only consider data

that are relevant to the nanomaterial(s) under evaluation, are sufficiently complete, and are

of appropriate quality to support the safety assessment.

The SCCS has also published a number of scientific opinions in the past few years on the

nano-form of different materials. These include 1,3,5-triazine, 2,4,6-tris[1,1’-biphenyl]-4-yl-

(ETH50)

(SCCS/1429/11, revision of 13/14 December 2011); zinc oxide (SCCS/1489/12

revision of 11 December 2012); titanium dioxide

(SCCS/1516/13, revision of 22 April 2014);

carbon black

(SCCS/1515/13, revision of 15 December 2015), 2,2’-methylene-bis-(6-(2H-

benzotriazol-2-yl)-4-(1,1,3,3-tetramethylbutyl)phenol), MBBT (SCCS/1460/11), silica

(SCCS/1545/15), hydroxyapatite (SCCS/1566/15); additional coatings for titanium dioxide

(SCCS/1580/16); and titanium dioxide in sprays (SCCS/1583/17). Styrene/Acrylates

copolymer (nano) and Sodium styrene/Acrylates copolymer (nano) (SCCS/1595/18);

Colloidal Silver (SCCS/1596/18); Solubility of Synthetic Amorphous Silica (SAS)

(SCCS/1606/19). These opinions can provide further information on the type of scientific

evidence needed in a safety dossier on nanomaterials intended for use as cosmetic

ingredients.

In general, a number of reviews have concluded that the existing risk assessment paradigm,

in use for conventional chemicals, should in principle be also applicable to engineered

nanomaterials. However, it has also been pointed out that the current testing methods may

need certain adaptations to take account of the special features of nanomaterials (Rocks et

al., 2008; SCENIHR, 2009; OECD, 2009c; SCCS, 2012; EC, 2012; ECHA, 2017; EFSA, 2018).

Special features of nanomaterials:

i. Due to high surface energies, nanoparticles have a tendency to stick together to form

agglomerates and aggregates, and/or bind with other moieties on the particle surface.

This particle behaviour can change in the presence of certain stabilising/dispersing agents.

Characterisation of nanomaterials, prior to and during a test, is therefore a key to ensuring

that results obtained are valid.

ii. Most of the currently available test methods were developed for conventional substances

that can be solubilised. In contrast, nanomaterials generally comprise insoluble or poorly

soluble nanoparticles that are dispersed in a test medium in the form of a nano-suspension

rather than a solution. The applied concentration of a nanomaterial may therefore drop

during the test due to particle agglomeration, sedimentation, binding with other moieties

_____________________________________________________________________________________________

in the medium, or sticking to the sides of the glass/plastic ware. This could lead to only a

partial or no exposure of the test systems during the test. Nanomaterials are known to

adsorb or bind different substances on their surfaces, including proteins (Šimon and Joner,

2008; Lynch and Dawson, 2008; Monopoli et al., 2012; Moore et al., 2015). They may also

bind other substances in the test medium and carry them into the exposed test systems,

leading to artefacts in the results.

iii. The toxicological hazards of chemical substances are currently measured and expressed in

terms of weight or volume units (such as mg/kg, or mg/l). These conventional metrics

may not be fully adequate to account for nanomaterial toxicity. It is therefore important

that tests on nanomaterials are not only evaluated in terms of weight/volume

concentration, but that results are also expressed in other dose-describing metrics, such

as particle number concentration, surface area etc.

iv. Due to the insoluble particulate nature, and the nano-dimensions, nanomaterials may show

an altered uptake and biokinetic profile in a biological system compared to equivalent

conventional forms e.g. transport of insoluble particles across biological membrane

barriers is not driven by concentration-gradient based diffusion partitioning, but by other

mechanisms such as endocytosis and/or active (energy-driven) uptake and transport.

v. Currently, there are uncertainties in regard to whether the endpoints identified by the

current testing methods will be sufficient to identify and characterise all the hazards that

may be associated with a nanomaterial.

3-6.8.3 REQUIRED INFORMATION FOR NANOMATERIALS

The information required by the SCCS for the evaluation of nanomaterials as cosmetic

ingredients is described in SCCS/1588/17 and SCCS/1611/19.

The following aspects deserve special attention:

i. Although most analytical methods used routinely for chemical substances have not yet

been validated for nanomaterials, a careful choice of mainstream method(s) should provide

sufficient means to gather adequate characterisation data for nanomaterials. The use of

more than one method generally adds more confidence to the measured values e.g. for

the measurement of particle size distribution, additional imaging by electron microscopy

has been recommended by both SCCS (SCCS/1611/19) and EFSA (EFSA, 2011b; EFSA,

2018).

ii. Where there is evidence for systemic absorption, further investigations are required to

confirm whether the absorbed material was in a nanoparticle form or in

solubilised/ionic/metabolised form. Where the absorption of nanoparticles cannot be ruled

out either by experimental measurements or justified on the basis of solubility/degradation

of the nanomaterial, the SCCS may apply a default approach and assume that 100% of

the absorbed material was in nano form.

iii. Surface modification/surface coating may bring about profound changes in a nanomaterial

in regard to certain physicochemical properties and potentially the toxic effects.

iv. Therefore, a full dataset would be preferable. As a minimum, in addition to safety data on

the core nanomaterial, the SCCS would require the following:

● Information/data on each material used for surface modification/coating of the

nanomaterial to indicate that it is safe for use in the intended cosmetic product.

● Data on physicochemical properties of the surface-modified/coated nanomaterial

to show that they have not significantly changed compared to either the same

material when uncoated, or with a different surface modification/coating that has

already been assessed safe by the SCCS.

● Data on dermal penetration, stability of the surface modification/coating, and

(photo)catalytic activity, where relevant.

_____________________________________________________________________________________________

3-6.9 HAIR DYES AND HAIR DYE COMPONENTS

In April 2003 the Commission, together with the Member States, agreed on a step-by-step

strategy to regulate all hair dyes listed as substances in cosmetic products. The main element

of the strategy was a tiered, modular approach, requiring industry to submit by certain

deadlines safety dossiers for hair dye components and possible mixtures. This strategy was

supported by SCCNFP (SCCNFP/0807/04) through its "Opinion on hair dyes without file

submitted", in which the experts clearly expressed the demand for a safety dossier for all hair

dyes, irrespective whether they had already been taken up in one of the annexes of the

cosmetic legislation. Differentiation was made between temporary, semi-permanent and

permanent hair dyes (SCCP/0959/05).

To ensure the safety of hair dye products, the Commission decided to ban all permanent,

semi-permanent and temporary hair dyes for which industry did not submit any safety files

and those for which the SCCP had given a negative opinion (IP/06/1047).

In 2013, the SCCS confirmed the views expressed in an earlier Memorandum (SCCP, 2006),

that hair dye substances which fulfil the criteria for classification as Skin Sens 1, H317

(according to CLP) may not be safe for consumers and that this is particularly so for hair dye

substances categorised as extreme and strong sensitisers (SCCS/1509/13).

3-6.9.1 MOS CALCULATIONS FOR HAIR DYE FORMULATIONS

Intermittent exposure and MoS calculations: hair dyes are not intended to be applied on a

daily basis. However, the MoS is calculated by dividing the PoD for daily application by the

SED for a single application. Although this approach can be debated, this is used as a

conservative approach.

Thus, the daily dose should not be averaged over the whole year (ECHA, 2012a).

3-6.9.2 ASSESSMENT OF OXIDATIVE HAIR DYE SUBSTANCES AND REACTION PRODUCTS

The SCCS is focused on the overall consumer health risk caused by ingredients as well as

products and intermediates of oxidative hair dyes formed during hair dyeing processes

(including their potential mutagenic/genotoxic/carcinogenic properties). The following

conclusions were drawn in the SCCS’s opinion on reaction products of oxidative hair dye

ingredients formed during hair dyeing processes (SCCS/1311/10):

- Precursors and couplers with a variety of substituents such as hydroxy, amino, imino

carbonyl, hydroxyethyl, hydroxyethoxy and alkyl groups were included.

- The use of oxidative hair dye formulations results in consumer exposure to precursors and

couplers as well as to their reaction products. Exposure to these reaction products is

considered generally lower compared to that from precursors and couplers since dimers

and trimers are formed with higher molecular weight. No exposure to intermediates or

self-coupling products was detected under experimental conditions. Therefore, in the risk

assessment of reaction products, toxicity is not considered a concern due to the low and

intermittent exposure (on average once per month).

- The dermal absorption rates in the in vitro skin penetration studies of the 14 representative

reaction products evaluated ranged from 3.27 to 717.79 ng/cm

(mean + 1 SD). This

corresponds to 1.9 to 416 µg absorbed dose (i.e. dose potentially bioavailable) per hair

dye application (i.e. 0.03 to 6.9 µg/kg bw).

- As no data were made available for sensitisation risk of the reaction products, this endpoint

was not specifically addressed.

_____________________________________________________________________________________________

- The use of (Q)SAR in the case of reaction products is of limited value so far since the

arylamine structure, a structural element of many hair dye precursors and reaction

products, is automatically identified as an alert. It is desirable to use or to develop in the

future SAR for in vivo genotoxicity which satisfies the OECD principles and has a known

applicability domain.

- Although for precursors, couplers and reaction products, positive results are commonly

observed in in vitro genotoxicity assays there is no clear evidence of genotoxicity in vivo

(in case in vivo data are available). It is possible that genotoxic effects can only be found

at concentrations where the N-acetylation (detoxifying) capacity of the cells is

overwhelmed, indicating that a ‘first-pass’ effect in skin could be taken into account for

risk assessment of the topically applied aromatic amines (Zeller and Pfuhler, 2014;

Nohynek et al., 2015).

- The structures of the primary intermediates and trimer molecules reveal that they contain

an aromatic secondary amino group, which if exposed to a nitrosating agent may form an

N-nitroso derivative (Lewis et al., 2013). Although, such transformation is theoretically

possible, no evidence was provided under real exposure conditions.

For all the above reasons, the SCCS performs the safety assessment of oxidative hair dyes

based on the toxicological evaluation of the ingredients (i.e. precursors and couplers) and not

the reaction products.

With regard to the animal testing ban for cosmetic ingredients, see Section 3-1 and the

scheme in Appendix 4.

3-6.10 COSMETIC INGREDIENTS FOR BABY AND CHILDREN PRODUCTS

In certain cases it may be necessary to calculate the MoS of cosmetic ingredients for a specific

subpopulation such as babies and children, e.g. exposure to leave-on cosmetic products

designed for application on the nappy area or products intended for children with a higher

sensitivity for certain endpoints. Also, differentiation between premature babies and full-

term neonates must be made since important structural and functional skin differences are

present. In particular, the barrier function in premature babies is impaired (Visscher et al.,

2015, 2020a, 2020b). Also, pH differences play a prominent role (Fluhr and Darlenski 2018;

Proksch 2018) which may be important for baby care products that are used often, such as

wet wipes (Rodriguez 2020; Gustin et al., 2020).

Here, only intact skin of full-term babies has been considered.

In light of potential differences in metabolism between newborns/infants up to six months

and adults, the question may be raised whether cosmetic ingredients would require a MoS

higher than 100 to cover exposure for these groups.

3-6.10.1 DEFINITIONS

“Children” are defined as developing human beings who are at various stages of immaturity

and maturation for up to nearly two decades, with age-dependent different susceptibilities

and sensitivities (Makri et al., 2004; Lemper et al., 2009) compared to adults.

Terms usually covered by the word “children” include:

- full-term neonate < 1 week

- newborn 1 week – 2 months

- early infant 2 – 6 months

- crawler/toddler 6 months – 2 years

- child/pre-adolescent 2 – 12 years

- adolescent 12 – 18 years

_____________________________________________________________________________________________

3-6.10.2 AGE-RELATED SUSCEPTIBILITIES/SENSITIVITIES

The calculation of the MoS for children was discussed when the question was raised whether

it would be advisable to adjust the default assessment factor of 100 for children by multiplying

this factor by the difference in Skin Surface Area over Body Weight ratio (SSA/BW) between

adults and children (SCCNFP/0557/02). In these calculations, the bodyweight values available

at that time were used. Afterwards, updated values became available (EFSA, 2012b).

The ratio between the SSA/BW ratios of children and adults changes from 0 to 10 years and

is as follows (Renwick, 1998):

2.3 at birth,

1.8 at 6 months,

1.6 at 12 months,

1.5 at 5 years,

1.3 at 10 years.

The ratio between the SSA/BW children of 0 to 1 year of age and that of adults is at maximum

2.3. A factor of 3.2 is generally applied by the WHO and also covers variability in human

kinetics (see Section 3-5.1.3). Consequently, the inter-individual variation in SSA/BW is

covered by the generally accepted default value of 100 for intact skin (Figure 10 in Section

3-5.1.3). However, for specific compounds under consideration the potential differences in

metabolism between newborns/infants up to six months and adults could require extra

consideration.

In general, the SCCS is of the opinion that there is no need for an additional UF for children

when intact skin is considered (SCCNFP/0557/02).

For more information on UFs, see 3-4.8.1.

Risk assessment in the specific case of “children” has been discussed for parabens as

preservatives in cosmetic products (SCCS/1446/11) and for phenoxyethanol

(SCCS/1575/16).

The rationale of additional UFs for different age groups beyond the usual factor of 100 has

been discussed in the scientific literature (e.g., Renwick et al., 1998 and 2000; Nielsen et al.,

2001; Makri et al., 2004; ECHA, 2012a).

A number of potential risk factors may exist for newborns and early infants. They are reviewed

in Annexes 2 and 4 of SCCS/1446/11. As dermal exposure in children is a topic of high

importance for several cosmetic substances, the most important points are summarised here.

An overview of potential risk factors for baby care products and their ingredients is also

available in Desmedt et al., 2014).

3-6.10.2.1 DERMAL EXPOSURE OF THE NEWBORN AND EARLY INFANT

- When born at full-term, the skin possesses all skin structures of adult skin, and

anatomically these structures do not undergo dramatic changes after birth. The dermal

absorption in skin of newborns is similar to that observed in adult skin, when the skin

is intact (see SCCS/1446/11) (Visscher et al., 2009 and 2015).

- Differences between newborns during their first weeks and months and adults are

described below:

(I) The surface area/body weight ratio (mentioned above) is 2.3-fold higher in newborns than

in adults, changing to 1.8- and 1.6-fold at 6 and 12 months, respectively. This is in general

covered by the intra-species factor of 10 (3.2 x 3.2) used in the calculation of MoS.

(II) Toxicokinetic parameters may differ between various age groups of children and adults

and can result in reduced metabolism, clearance and/or longer half-life that might either

The considerations in this section refer to neonates born at full-term and not to premature babies still under

medical care.

_____________________________________________________________________________________________

increase or decrease the potential risk of an adverse reaction in newborns, depending on the

substance (Renwick et al., 2000; Nielsen et al., 2001, Felter et al., 2015). For the CYP450s

in the liver, lower activities in newborns/early infants as compared to adults have been

described (Johnson, 2003). These data suggest that the extent of bioactivation or metabolic

detoxification in children between one and ten years will in generally be lower than that in

adults. It is also known that detoxification of xenobiotic substances or metabolites by phase

II enzymes may be lower in newborns and infants compared to adults due to yet incomplete

development of Xenobiotic Metabolising Enzymes (XME) in the liver (e.g., UDP

GlucuronosylTransferase-1 (UGT1A1) and some esterases; see SCCS/1446/11). Therefore,

depending on the cosmetic ingredient in question, the balance between activating and

inactivating XME activities may be crucial for systemic exposure and should be considered

case by case. In general, however, it is assumed that a specific assessment factor for age-

related differences in toxicokinetics is not required (SCCS/1446/11). With respect to skin

metabolism, it is recognised that some metabolic enzymes seem to be less expressed in the

skin of children, in particular under the age of 1 year. Hence, neonates, newborns and early

infants might have higher internal exposure to certain cosmetic ingredients after dermal

application than adults. For a sound risk assessment, relevant human data regarding

metabolism are necessary. These data could for instance be gained by an approach combining

in vitro data on the metabolism of the cosmetic ingredient under investigation and PBPK/PBTK

modelling. For such toxicokinetic modelling of the biotransformation in humans of different

age groups, relevant in vitro data regarding phase I and phase II biotransformation are

needed both in human skin and liver (SCCS/1446/11).

(III) In-use conditions of topical products should be considered in exposure-based risk

assessment of the finished product. It should be noted that no comprehensive exposure data

for newborns and early infants, representative for Europe, are available in the open literature.

CoE is preparing aggregate exposure data for babies and children for different baby care

cosmetics used in Europe (more information, see Table A.7, Appendix 7).

Some information is available for the Netherlands at the RIVM, ConsExpo Fact Sheet (2006).

Data for French children have been published by Ficheux et al., 2017, 2019. Exposure data

for wipes used for Korean babies are available (Lee et al., 2017); also for the USA, DE and

UK,deterministic as well as probabilistic modelling has been carried out to determine the

transfer of wipes in babies and children (Dey et al., 2016a). Data for disposable diapers are

available from the same authors (Dey et al., 2016b).

(IV) The nappy area: the skin barrier function in the nappy area and non-nappy regions are

indistinguishable at birth but show differential behaviour over the first 14 days, with the

nappy region having a higher pH and increased hydration. With respect to skin hydration in

the nappy zone, newborns tend to have a somewhat higher water content in the horny layer

than observed for early infants and crawlers/toddlers up to one year. Also, the variations in

water content are higher. Skin pH is usually between 5-6, which is similar to the skin pH

measured for adult skin. However, the nappy area is susceptible to inflammation and the

buffering capacity is compromised (nappy dermatitis). This results in episodic acute skin

inflammation (mean duration 2 to 3 days) caused as well by physical, chemical andenzymatic

microbial factors in the nappy environment, for example acute skin inflammation of the nappy

zone occurs during changes in diet (breast feeding, bottle feeding, solid food) and may occur

in particular between 6-12 months of age.

See below for cosmetic products used in the nappy area.

(V) Susceptibility against microorganisms: this is in particular the case in the nappy area and

is a consequence of changes in the barrier function when the skin is damaged. Therefore,

baby cosmetics should be adequately preserved (as is the case for all cosmetics) and

formulated with an appropriate buffered pH.

With respect to points (I) to (III), there is generally no need for an additional assessment

factor for children when intact skin is involved. However, an additional assessment

factor might be relevant when the skin in the nappy area is damaged and substance-

_____________________________________________________________________________________________

100

specific data clearly demonstrate that inter-individual variability would result in a value higher

than the default value of 10.

3-6.10.2.2 COSMETIC PRODUCTS USED IN THE NAPPY AREA

In the nappy area, special circumstances are present resulting from the close confining

clothes and nappies, uncontrolled urination and defecation and resulting problems with

potential damage of the skin in the nappy zone. Modern nappy technology has shown to

provide increasingly good skin compatibility, leading to a decline in the frequency and severity

of nappy dermatitis. In silico modeling of skin under the diaper has shown that healthy

diapering practices will ensure there is no significant impact on skin health and barrier

properties (Staadatmand et al., 2017). However, irritant nappy dermatitis cannot be

completely avoided and might have an impact on dermal absorption of substances.

As cosmetic products are meant to be used on intact skin, medical consultation is necessary

in the case of real skin damage and pharmaceutical products (and not cosmetics!) should be

used.

For the development of baby cosmetic products and the safety evaluation of the products

intended to be used in the nappy area, the potential impact of irritation on dermal absorption

of the ingredients needs to be considered by the safety assessor. It is known that the physico-

chemical properties of the substances under consideration also play a role.

A tiered quantitative approach to take the potential for diaper rash into consideration when

doing a safety evaluation for products used in the nappy area has been proposed by Felter et

al. (Felter et al., 2017).

3-6.11 SUBSTANCES WITH VERY LOW DERMAL ABSORPTION

In the case where a cosmetic ingredient is a substance with a very low dermal absorption

{see Section 3-3.5.1.1(c)}, some studies could be waived since systemic exposure via dermal

absorption is expected to be minimal. In such a case, the following minimum set of data

should be made available in order to assess the safety of cosmetic ingredients with very low

bioavailability:

- Experimentally determined physicochemical data

- Local toxicity

- Mutagenicity/Genotoxicity

- High quality in vitro dermal absorption study, according to the SCCS Basic Criteria {3-

3.5.1.1 (b}.

In these cases, the experimental mean value will be used for decision making.

3-7 FURTHER REMARKS FOR APPLICANTS

- When preparing a safety dossier, it would be useful if Applicants follow the same format

as adopted in the SCCS opinions (example given in Appendix 3).

- Whenever study results are submitted, a declaration should be made that the tests

involved were conducted using a cosmetic ingredient with a comparable purity/impurity

profile and physical and chemical characteristics of that to be included in the finished

cosmetic product.

- For multi-constituent natural ingredients, with variable composition, it is essential that

Applicants provide clearly defined specifications in view of the range of variability of the

components e.g. batch-to-batch.

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101

- Stability of the test substance under experimental conditions is of prime importance for

the interpretation of test results.

- The stability of the test material under conditions of use should also be reported.

- The Applicant should ensure that files submitted for evaluation are complete and signed.

Data should be obtained by means of studies conducted in accordance with test guidelines

reported in Regulation (EC) No 440/2008 (2008/440/EC) and amending ATP (Adaptation

to Technical and scientific Progress) Regulations, as well as the OECD test guidelines, and

complying with the principles of Good Laboratory Practice (GLP). All possible deviations

from validated methods or from GLP must be indicated, explained and scientifically

justified. There may be cases for which it is either not necessary or technically not possible

to provide some of the information mentioned above: in such cases a scientific justification

must be given by industry and/or relevant agencies.

- Together with the relevant experimental investigations, the following information should be

provided:

- for in vivo studies: the study date (whether in line with the Cosmetic

Regulation) and/or the regulatory context for which the study has been

performed;

- any report on epidemiological and/or observational experiences

(cosmetovigilance data);

- an appraisal of all relevant published literature, along with a description of the

bibliographical methods used; any information from "grey material" available.

Any other relevant findings by the Applicant and/or other industry/agencies,

should also be transmitted to the Commission for review.

- In their dossiers, the Applicants should indicate whether they consider any of the

data/tables/substances names, etc. confidential (typically impurities etc.) for commercial

reasons and provide relevant codes that can be used by the SCCS to anonymise the

confidential information.

- Safety data must relate to the same form of ingredients as present in a product for final

use keeping in mind that the formulation or preparation of the final product may change

the nature of the ingredients (e.g. permanent hair dye preparation).

- In case there is a negative SCCS Opinion, the Applicant must consider whether sufficient

new and relevant information is available to justify a resubmission. When a dossier is

resubmitted, it is mandatory to provide it in the form of a full dossier (including references)

and clearly indicate what is new compared to the previous submission(s).

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102

4. REFERENCE LIST

Regulations and Decisions from the Commission are ordered by year.

67/548/EEC - Council Directive 67/548/EEC of 27 June 1967 on the approximation of laws,

regulations and administrative provisions relating to the classification, packaging and labelling

of dangerous substances. Official Journal P 196, 16/08/1967 p.1.

76/768/EEC - Council Directive 76/768/EEC of 27 July 1976 on the approximation of the

laws of the Member States relating to cosmetic products. Official Journal L 262, 27/09/1976

p.169.

78/45/EEC - Commission Decision 78/45/EEC of 19 December 1977 establishing a Scientific

Committee on Cosmetology. Official Journal L 13, 17/01/1978 p.24.

87/18/EEC - Council Directive 87/18/EEC of 18 December 1986 on the harmonisation of

laws, regulations and administrative provisions relating to the application of the principles of

good laboratory practice and the verification of their applications for tests on chemical

substances. Official Journal L 15, 17/01/1987 p.29.

93/35/EEC - Council Directive 93/35/EEC of 14 June 1993 amending for the sixth time

Directive 76/768/EEC on the approximation of the laws of the Member States relating to

cosmetic products. Official Journal L 151, 23/06/1993 p.32.

96/335/EC - Commission Decision of 8 May 1996 establishing an inventory and a common

nomenclature of ingredients employed in cosmetic products. Official Journal L 132,

01/06/1996 p.1

2000/60/EC – Directive 2000/60/EC of the European Parliament and of the Council

establishing a framework for the Community action in the field of water Official Journal L 327,

22/12/ 2000pp.1-73.

2002/178/EC - Regulation (EC) No 178/2002 of the European Parliament and of the Council

of 28 January 2002 laying down the general principles and requirements of food law,

establishing the European Food Safety Authority and laying down procedures in matters of

food safety, Official Journal L 31, 1.2.2002, p. 1

2003/15/EC - Directive 2003/15/EC of the European Parliament and of the Council of 27

February 2003 amending Council Directive 76/768/EEC on the approximation of the laws of

the Member States relating to cosmetic products. Official Journal L66, 11/03/2003 p.26.

2004/210/EC - Commission Decision of 3 March 2004 setting up Scientific Committees in

the field of consumer safety, public health and the environment. Official Journal L 66,

04/03/2004 p.45.

2006/257/EC - Commission Decision 2006/257/EC of 9 February 2006 amending Decision

96/335/EC establishing an inventory and a common nomenclature of ingredients employed

in cosmetic products. Official Journal L 97, 05/04/2006 p.1.

2006/647/EC - Commission Recommendation of 22 September 2006 on the efficacy of

sunscreen products and the claims made relating thereto (notified under document number

C(2006) 4089), Official Journal L 265, 26.9.2006, pp. 39–43

2006/1907/EC - Regulation (EC) No 1907/2006 of the European Parliament and of the

Council of 18 December 2006 concerning the Registration, Evaluation, Authorisation and

_____________________________________________________________________________________________

103

Restriction of Chemicals (REACH), establishing a European Chemicals Agency, amending

Directive 1999/45/EC and repealing Council Regulation (EEC) No 793/93 and Commission

Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC and Commission

Directives 91/155/EEC, 93/67/EEC, 93/105/EC and 2000/21/EC. Official Journal L 396,

30/12/2006, p.1. Corrigendum in Official Journal L 136, 29/05/2007, p.3.

2008/440/EC - Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test

methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the

Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

Official Journal L 142, 31/05/2008 p.1.

2008/721/EC - Commission Decision 2008/721/EC of 5 September 2008 setting up an

advisory structure of Scientific Committees and experts in the field of consumer safety, public

health and the environment and repealing Decision 2004/210/EC. Official Journal L 241,

10/09/2008 p.21.

2008/771/EC - Commission regulation (EC) No 771/2008 of 1 August 2008 laying down the

rules of organisation and procedure of the Board of Appeal of the European. Official Journal L

206, 2/08/2008 p.5.

2008/1272/EC - Regulation (EC) No 1272/2008 of the European Parliament and the Council

of 16 December 2008 on classification, labelling and packaging of substances and mixtures,

amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation

(EC) No 1907/2006. Official Journal L 353, 31/12/2008 p.1.

2009/146/EC - Commission Decision 2009/146/EC of 19 February 2009 on the appointment

of the members and advisors of the Scientific Committees and the Pool set up by Decision

2008/721/EC. Official Journal L 49, 20/02/2009 p.33.

2009/1107/EC - Regulation (EC) 1107/2009 of the European Parliament and of the Council

of 21 October 2009 concerning the placing of plant protection products on the market and

repealing Council Directives 79/117/EEC and 91/414/EEC. Official Journal L 309, 24

November 2009 pp.1-50

2009/1223/EC - Regulation (EC) No 1223/2009 of the European Parliament and of the

Council of 30 November 2009 on cosmetic products (recast). Official Journal L 342,

22/12/2009 p.59.

2011/696/EU – Commission Recommendation of 18 October 2011 on the definition of

nanomaterials. Official Journal L 275, 18/10/2011 p.38.

2012/528/EC - Regulation (EC) 528/2012 of the European Parliament and of the Council of

22 May 2012 concerning the making available on the market and use of biocidal products.

Official Journal L 167, 27/6/2012 pp.1-123.

2015/5383/EC - Commission Decision C(2015) 5383 final of 7 August 2015 on establishing

Scientific Committees in the field of public health, consumer safety and the environment.

https://ec.europa.eu/health/scientific_committees/docs/call_2015_5383_decision_with_ann

exes_en.pdf

Aberdam E, Petit I, Sangari L, Aberdam D, Induced pluripotent stem cell-derived limbal

epithelial cells (LiPSC) as a cellular alternative for in vitro ocular toxicity testing, PLoS One

12(6):e0179913 (2017). doi: 10.1371/journal.pone.0179913

Adler, S.; Basketter, D.; Creton, S.; Pelkonen, O.; van Benthem, J.; Zuang, V.; Andersen,

KE.; Angers-Loustau, A.; Aptula, A.; Bal-Price, A.; Benfenati, E.; Bernauer, U.; Bessems, J.;

_____________________________________________________________________________________________

104

Bois, FY.; Boobis, A.; Brandon, E.; Bremer, S.; Broschard, T.; Casati, S.; Coecke, S.; Corvi,

R.; Cronin, M.; Daston, G.; Dekant, W.; Felter, S.; Grignard, E.; Gundert-Remy, U.;

Heinonen, T.; Kimber, I.; Kleinjans, J.; Komulainen, H.; Kreiling, R.; Kreysa, J.; Leite, SB.;

Loizou, G.; Maxwell, G.; Mazzatorta, P.; Munn, S.; Pfuhler, S.; Phrakonkham, P.; Piersma,

A.; Poth, A.; Prieto, P.; Repetto, G.; Rogiers, V.; Schoeters, G.; Schwarz, M.; Serafimova,

R.; Tähti, H.; Testai, E.; van Delft, J.; van Loveren, H.; Vinken, M.; Worth, A.; Zaldivar, J.M.

Alternative (non-animal) methods for cosmetics testing: current status and future prospects-

2010. Arch Toxicol 85(5): 367-485 (2011). doi: 10.1007/s00204-011-0693-2

Åkerlund, E., Islam, M. S., McCarrick, S., Alfaro-Moreno, E. & Karlsson, H. L. Inflammation

and (secondary) genotoxicity of Ni and NiO nanoparticles. Nanotoxicology 13:8 1060–1072

(2019). doi: 10.1080/17435390.2019.1640908

Alépée N., Barroso J., De Smedt A., De Weverd B., 2, Hibatallah J., Klaric M., Mewes K.R.,

Millet M., Pfannenbecker U., Tailhardat M., Templier M., P. McNamee, Use of HPLC/UPLC-

spectrophotometry for detection of formazan in in vitro Reconstructed human Tissue (RhT)-

based test methods employing the MTT-reduction assay to expand their applicability to

strongly coloured test chemicals, Toxicol in vitro, 29(4): 741–761 (2015). doi:

10.1016/j.tiv.2015.02.005

Aljallal M. Investigation of in Silico Modelling to Predict the Human Health Effects of

Cosmetics Ingredients. Doctoral thesis, Liverpool John Moores University (2020). doi:

10.24377/LJMU.t.00012139

Al-Zoughool, M., Bird, M., Rice, J., Baan, R. A., Billard, M., Birkett, N. & Zielinski, J. M.

Development of a database on key characteristics of human carcinogens. Journal of

Toxicology and Environmental Health, Part B, 22(7-8), 264-287. (2019). doi:

10.1080/10937404.2019.1642593

Andres E., Sá-RochaV.M., Barrichello C., Haupt T., Ellis G., Natsch A. The sensitivity of the

KeratinoSens™ assay to evaluate plant extracts: A pilot study, Toxicol in vitro 27: 1220–

1225 (2013). doi: 10.1016/j.tiv.2013.02.008

Anton, D., Burckel, H., Josset, E., Noel, G., 2015. Three-dimensional cell culture: A

breakthrough in vivo. Int. J. Mol. Sci. 16: 5517–5527 (2015). doi: 10.3390/ijms16035517

Api, A. M., Basketter, D., Bridges, J., Cadby, P., Ellis, G., Gilmour, N., Greim, H., Griem, P.,

Kern, P., Khaiat, A., O’Brien, J., Rustemeyer, T., Ryan, C., Safford, B., Smith, B., Vey, M. &

White, I. R. Updating exposure assessment for skin sensitization quantitative risk assessment

for fragrance materials. Regul Toxicol Pharmacol 118: 104805 (2020). doi :

10.1016/j.yrtph.2020.104805

Ashby, J. & Tennant, R. W. Chemical structure, Salmonella mutagenicity and extent of

carcinogenicity as indicators of genotoxic carcinogenesis among 222 chemicals tested in

rodents by the U.S. NCI/NTP. Mutat Res Toxicol 204(1): 17–115 (1988). doi: 10.1016/0165-

1218(88)90114-

Ates G., Steinmetz FP., Doktorova TY., Madden JC., Rogiers V. Linking existing in vitro dermal

absorption data to physicochemical properties: Contribution to the design of a weight-of-

evidence approach for the safety evaluation of cosmetic ingredients with low dermal

bioavailability. Regul Toxicol Pharmacol 76: 74-78 (2016). doi: 10.1016/j.yrtph.2016.01.015

Ates G., Mertens B., Heymans A., Verschaeve L., Milushev D., Vanparys P., Roosens NHC.,

De Keersmaecker SCJ., Rogiers V., Doktorova TY. A novel genotoxin-specific qPCR array

based on the metabolically competent human HepaRG™ cell line as a rapid and reliable tool

for improved in vitro hazard assessment. Arch Toxicol 92(4):1593-1608 (2018). doi:

10.1007/s00204-018-2172-5

_____________________________________________________________________________________________

105

Autier P., Boniol M., Severi G., Doré J-F. Quantity of sunscreen used by European students.

Brit J Dermatol 144(2): 288-291 (2001). doi: 10.1046/j.1365-2133.2001.04016.x

Autier P., Boniol M., Doré J-F. Sunscreen use and increased duration of intentional sun

exposure: still a burning issue. Int J Cancer 121(1): 1-5 (2007). doi: 10.1002/ijc.22745

Averbeck D., Moustacchi E. Genetic effect of 3-Carbethoxypsoralen angelicin, psoralen and

8-methoxypsoralen plus 365 nm irradiation in Saccharomyces cerevisiae: induction of

reversions, mitotic crossing-over, gene conversions and cytoplasmic “petite” mutations.

Mutat Res 68(2): 133-148 (1979). doi: 10.1016/0165-1218(79)90141-1

Bakhtyari, N. G., Raitano, G., Benfenati, E., Martin, T. & Young, D. Comparison of in silico

models for prediction of mutagenicity. J Environ Sci Heal - Part C Environ Carcinog Ecotoxicol

Rev 31(1): 45–66 (2013). doi: 10.1080/10590501.2013.763576

Balls M. and Fentem J.H. Progress toward the validation of alternative tests. Altern Lab Anim.

25: 33-43 (1997).

Baltazar, M. T., Cable, S., Carmichael, P. L., Cubberley, R., Cull, T., Delagrange, M., Dent,

M. P., Hatherell, S., Houghton, J., Kukic, P., Li, H., Lee, M.-Y., Malcomber, S., Middleton, A.

M., Moxon, T. E., Nathanail, A. V, Nicol, B., Pendlington, R., Reynolds, G., Reynolds, J., White,

A. & Westmoreland, C. A Next-Generation Risk Assessment Case Study for Coumarin in

Cosmetic Products. Toxicol Sci 176(1): 236–252 (2020). doi: 10.1093/toxsci/kfaa048

Baptista, P.M., Moran, E.C., Vyas, D., Ribeiro, M.H., Atala, A., Sparks, J.L., Soker, S., Fluid

Flow Regulation of Revascularization and Cellular Organization in a Bioengineered Liver

Platform. Tissue Eng. Part C Methods 22: 199–207 (2016). doi: 10.1089/ten.TEC.2015.0334

Basketter DA., Andersen KE., Lidén C., Van Loveren H., Boman A., Kimber I., Alanko K.,

Berggren E. Evaluation of the skin sensitising potency of chemicals by using the existing

methods and considerations of relevance for elicitation. Contact Dermatitis 52(1): 39-43

(2005). doi: 10.1111/j.0105-1873.2005.00490.x

Bech-Thomsen N., Wulf HC. Sunbathers’ application of sunscreen is probably inadequate to

obtain the sun protection factor assigned to the preparation. Photodermatol. Photoimmunol.

Photomed. 9(6): 242-244 (1993).

Beloqui A., des Rieux A., Préat V., Mechanisms of transport of polymeric and lipidic

nanoparticles across the intestinal barrier, Adv Drug Deliv Rev.106 (Pt B):242-255 (2016).

doi: 10.1016/j.addr.2016.04.014

Bemis, J. C. & Heflich, R. H. In vitro mammalian cell mutation assays based on the Pig-a

gene: A report of the 7th International Workshop on Genotoxicity Testing (IWGT) Workgroup.

Mutation Research - Genetic Toxicology and Environmental Mutagenesis vol. 847: 403028

(2019). doi: 10.1016/j.mrgentox.2019.03.001

Benfenati, E. E-Book: Theory, guidance and applications on QSAR and REACH, ISBN 978-

88-902405-4-6; http://home.deib.polimi.it/gini/papers/orchestra.pdf, (2012).

Benigni R., Bossa C., Jeliazkova N., Netzeva T., Worth A. Structure alerts for carcinogenicity,

and the Salmonella assay system: A novel insight through the chemical relational databases

technology. Mutation Research - Reviews in Mutation Research vol. 659 248–261 (2008). doi:

10.1016/j.mrrev.2008.05.003

Berggren, E., White, A., Ouedraogo, G., Paini, A., Richarz, A. N., Bois, F. Y., Exner, T., Leite,

S., Grunsven, L. A. va., Worth, A. & Mahony, C. Ab initio chemical safety assessment: A

_____________________________________________________________________________________________

106

workflow based on exposure considerations and non-animal methods. Comput Toxicol 4: 31–

44 (2017). doi: 10.1016/j.comtox.2017.10.001

Bergström MA., Ott H., Carlsson A., Neis M., Zwadlo-Klarwasser G., Jonsson CA., Merk HF.,

Karlberg AT., Baron JM. A skin-like cytochrome P450 cocktail activates prohaptens to contact

allergenic metabolites. J Invest Dermatol 127(5):1145-53 (2007). doi:

10.1038/sj.jid.5700638

Bernard A., Houssin A., Ficheux AS., Wesolek N., Nedelec AS., Bourgeois P., Hornez N.,

Batardière A., Misery L., Roudota AC., Consumption of hair dye products by the French women

population: Usage pattern and exposure assessment, Food Chem Toxicol. 88:123-32 (2016).

doi: 10.1016/j.fct.2016.01.002

Bernard A., Dornic N., Roudt AC., Ficheux AS., Probabilistic exposure assessment to face

and oral care cosmetic products by the French population, Food Chem Toxicol. 111:511-524

(2018). doi: 10.1016/j.fct.2017.11.056

Berry C. The failure of rodent carcinogenesis as a model for Man. Toxicol Res (Camb) 7:

553–557 (2018). doi: 10.1039/c7tx00283a

Bessems Loizou G., Krishnan K., Clewell H.J., Bernasconi C., Bois F., Coecke S., Collnot

E.M., Diembeck W., Farcal L.R., Geraets L., Gundert-Remy U., Kramer N, Küsters G, Leite

SB, Pelkonen O.R., Schröder K., Testai E., Wilk-Zasadna I., Zaldívar-Comenges J.M. PBTK

modelling platforms and parameter estimation tools to enable animal-free risk assessment.

Recommendations from a joint EPAA – EURL ECVAM ADME workshop. Regul Toxicol

Pharmacol 68(1):119–139 (2014). doi: 10.1016/j.yrtph.2013.11.008

Bhatia S., Schultz, T., Roberts, D., Shen, J., Kromidas, L., Api, A.M. Comparison of Cramer

classification between toxtree, the OECD QSAR toolbox and expert judgment. Regul Toxicol

Pharmacol. 71(1):52-62 (2015). doi: 10.1016/j.yrtph.2014.11.005

Biesterbos J.W.H., Dudzina T., Delmaar C. J.E., Bakker M.I., Russel F.G.M., von Goetz N.,

Scheepers P.T.J., Roeleveld N. Usage patterns of personal care products: Important factors

for exposure assessment. Food Chem Toxicol 55:8–17 (2013). doi:

10.1016/j.fct.2012.11.014

Boobis A., Brown P., Cronin MTD., Edwards J., Galli CL., Goodman J., Jacobs A., Kirkland D.,

Luijten M., Marsaux C., Martin M., Yang C., Hollnagel HM., Origin of the TTC values for

compounds that are genotoxic and/or carcinogenic and an approach for their re-evaluation,

Critical Rev Toxicol, 47(8):705-727 (2017). doi: 10.1080/10408444.2017.1318822

Braakhuis H.M., Park MV., Gosens I., De Jong WH. and Cassee FR., Physicochemical

characteristics of nanomaterials that affect pulmonary inflammation, Part Fibre Toxicol 11:18-

44 (2014). doi: 10.1186/1743-8977-11-18

Brandsma, I., Moelijker, N., Derr, R. & Hendriks, G. Aneugen Versus Clastogen Evaluation

and Oxidative Stress-Related Mode-of-Action Assessment of Genotoxic Compounds Using the

ToxTracker Reporter Assay. Toxicol Sci 177: 202–213 (2020). doi: 10.1093/toxsci/kfaa103

Bremmer HJ., Prud'Homme de Lodder LCH., van Engelen JGM. Cosmetics Fact Sheet to

assess the risks for the consumer. Updated version for ConsExpo 4. RIVM Report 320104

001/2006 (2006a). Available at: https://www.rivm.nl/bibliotheek/rapporten/320104001.pdf

(Consulted December 2020).

Bremmer HJ., Prud'Homme de Lodder LCH., van Engelen JGM. General Fact Sheet - Limiting

conditions and reliability, ventilation, room size, body surface area. Updated version for

_____________________________________________________________________________________________

107

ConsExpo 4, RIVM report 320104002/2006 (2006b). Available at:

https://www.rivm.nl/bibliotheek/rapporten/320104002.pdf.(Consulted December 2020).

Brendler-Schwaab S., Czich A., Epe B., Gocke E., Kaina B., Müller L., Pollet D., Utesch D.

Photochemical genotoxicity: principles and test methods - Report of a GUM task force. Mutat

Res 566(1): 65-91 (2004). doi: 10.1016/s1383-5742(03)00052-8

Brown JS., Gordon T., Price O., Asgharian B., Thoracic and respirable particle definitions for

human health risk assessment; Part Fibre Toxicol., Apr 10;10:12 (2013). doi: 10.1186/1743-

8977-10-12

Bruynzeel DP. The European Task Force for Photopatch Testing Photopatch testing: a

consensus methodology for Europe. J Eur Acad Dermatol 18(6): 679-682 (2004). doi:

10.1111/j.1468-3083.2004.01053.x

Calafat AM., Needham LL. What additional factors beyond state-of-the art analytical

methods are needed for optimal generation and interpretation of biomonitoring data? Environ

Health Perspect 117(10), 1481-1485 (2009). doi: 10.1289/ehp.0901108

Callegaro G., Corvi R., Salovaara S., Urani C., Stefanini FM. Relationship between increasing

concentrations of two carcinogens and statistical image descriptors of foci morphology in the

cell transformation assay. J Appl Toxicol.37(6):709-720 (2017). doi: 10.1002/jat.3419

Carthew P., Griffiths H., Keech S., Hartop P. Safety assessment for hairspray resins: risk

assessment based on rodent inhalation studies. Inhal Toxicol 14(4), 401–416 (2002). doi :

10.1080/08958370252871023

Carthew, P., Clapp, C. & Gutsell, S. Exposure based waiving: The application of the

toxicological threshold of concern (TTC) to inhalation exposure for aerosol ingredients in

consumer products. Food Chem Toxicol 47: 1287–1295 (2009). doi:

0.1016/j.fct.2009.02.024

CEN (Comité Européen de Normalisation) Workplace Atmospheres: Size Fraction Definitions

for Measurement of Airborne Particles in the Workplace, CEN Standard EN 481 (1993).

Chen, C., Soto-Gutierrez, A., Baptista, PM., Spee, B., Biotechnology Challenges to In vitro

Maturation of Hepatic Stem Cells. Gastroenterology (scholar), 2018.

Chetelat A., Albertini S., Dresp H., Strobel R., Gocke E. Photomutagenesis test development.

Part I. 8-Methoxypsoralen, chloropromazine and sunscreen compounds in bacterial and yeast

assays. Mutat Res 292(3): 241-250 (1993a). doi: 10.1016/0165-1161(93)90027-w

Chetelat A., Dresp JH., Gocke E. Photomutagenesis test development. Part II. 8-

Methoxypsoralen, chlorpromazine and sunscreen compounds in chromosomal aberration

assays using CHO cells. Mutat Res 292(3): 251-258 (1993b). doi: 10.1016/0165-

1161(93)90028-x

Chetelat A., Albertini S., Gocke E. The photomutagenicity of fluoroquinolones in tests for

gene mutation, chromosomal aberration, gene conversion and DNA breakage (Comet assay).

Mutagenesis 11(5), 497-504 (1996). doi: 10.1093/mutage/11.5.497

Chiu WA, Guyton KZ, Martin MT, Reif DM, Rusyn I. Use of high-throughput in vitro toxicity

screening data in cancer hazard evaluations by IARC Monograph Working Groups. ALTEX

35(1):51-64 (2018). doi: 10.14573/altex.1703231

_____________________________________________________________________________________________

108

COC 2020 Committee on Carcinogenicity of Chemicals in Food, Consumer Products and the

Environment. COC Guidance Statement G05 – version 2-0 - Defining a Point of Departure and

Potency Estimates in Carcinogenic Dose Response.

Coecke S., Pelkonen O., Leite SB. Toxicokinetics as a key to the integrated toxicity risk

assessment based primarily on non-animal approaches. Toxicol in vitro 27(5):1570– 1577

(2013). doi: 10.1016/j.tiv.2012.06.012

COLIPA Guidelines: 'Consumer exposure to cosmetic ingredients', BB-97/007, 1997.

COM (UK Committee on Mutagenicity of Chemicals in Food, Consumer Products and the

Environment). Guidance on a Strategy for genotoxicity Testing and mutagenic hazard

assessment of Chemical substances. Available at:

https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_

data/file/315800/in_vivo_testing_of_genotoxicity_of_chemicals.pdf. Consulted December

2020.

COM (UK Committee on Mutagenicity of Chemicals in Food, Consumer Products and the

Environment) A Statement on photogenotoxicity testing, 2013, COM/13/S1. Available at:

https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/315914/sta

tement on_photogenotoxicity.pdf

COM (UK Committee on Mutagenicity of Chemicals in Food, Consumer Products and the

Environment) Communication from the Commission to the European Parliament and the

Council on the animal testing and marketing ban and on the state of play in relation to

alternative methods in the field of cosmetics, 2013, COM/2013/135

Comiskey D , A.M. Api, C. Barratt , E.J. Daly, G. Ellis, C. McNamara, C. O’Mahony, S.H.

Robison, B. Safford , B. Smith , S. Novel database for exposure to fragrance ingredients in

cosmetics and personal care products, Regul Toxicol Pharmacol 72(3): 660–672 (2015). doi:

10.1016/j.yrtph.2015.05.012

Corvi R, Madia F., Guyton KZ., Kasper P., Rudel R., Colacci A., Kleinjans J., Jennings P.

Moving forward in carcinogenicity assessment: Report of an EURL ECVAM/ESTIV workshop.

Toxicol In vitro 45(Pt 3):278-286 (2017). doi: 10.1016/j.tiv.2017.09.010

Cottrez F, Boitel E, Auriault C, Aeby P, Groux H. Genes specifically modulated in sensitized

skins allow the detection of sensitizers in a reconstructed human skin model. Development of

the SENS-IS assay. Toxicol In vitro 29(4):787-802 (2015). doi: 10.1016/j.tiv.2015.02.012

Cottrez F., Boitel E, Ourlin JC, Peiffer JL, Fabre I, Henaoui IS, Mari B, Vallauri A, Paquet A,

Barbry P, Auriault C, Aeby P, Groux H. SENS-IS, a 3D reconstituted epidermis based model

for quantifying chemical sensitization potency: Reproducibility and predictivity results from

an inter-laboratory study. Toxicol In vitro 32:248-260 (2016). doi:

10.1016/j.tiv.2016.01.007

Council of Europe. European Pharmacopoeia (Ph. Eur.) 10th ed. Strasbourg: Council of

Europe; General Notices, (2020).

Danish Environmental Protection Agency, Survey of chemical substances in consumer

products No. 151 (2016).

Dean S.W., Lane M., Dunmore RH., Ruddock SP., Martin CN., Kirkland DJ., Loprieno N.

Development of assays for the detection of photogenotoxicity of chemicals during exposure

to UV light. Part 1. Assay development. Mutagenesis 6(5): 335-341 (1991). doi:

10.1093/mutage/6.5.335

_____________________________________________________________________________________________

109

Dearfield K.L., Thybaud V., Cimino M.C., Custer L., Czich A., Harvey J.S., Hester S., Kim

J.H., Kirkland D., Levy D.D., Lorge E., Moore M.M., Ouedraogo-Arras G., Schuler M., Suter

W., Sweder K., Tarlo K., van Benthem J., van Goethem F. and Witt K.L. Follow-Up Actions

from Positive Results of In vitro Genetic Toxicity Testing. Environ Mol Mutagen 52(3): 177-

204 (2011). doi: 10.1002/em.20617

Dearfield, K. L., Gollapudi, B. B., Bemis, J. C., Benz, R. D., Douglas, G. R., Elespuru, R. K.,

Johnson, G. E., Kirkland, D. J., LeBaron, M. J., Li, A. P., Marchetti, F., Pottenger, L. H., Rorije,

E., Tanir, J. Y., Thybaud, V., van Benthem, J., Yauk, C. L., Zeiger, E. & Luijten, M. Next

generation testing strategy for assessment of genomic damage: A conceptual framework and

considerations. Environmental and Molecular Mutagenesis (2017). doi:10.1002/em.22045.

doi: 10.1002/em.22045

Delrue N., Sachana M., Sakuratani Y., Gourmelon A., Leinala E., Diderich R., The adverse

outcome pathway concept: A basis for developing regulatory decision-making tools. ATLA

44(5):417-429 (2016). doi: 10.1177/026119291604400504

Dertinger, S. D., Totsuka, Y., Bielas, J. H., Doherty, A. T., Kleinjans, J., Honma, M.,

Marchetti, F., Schuler, M. J., Thybaud, V., White, P. & Yauk, C. L. High information content

assays for genetic toxicology testing: A report of the International Workshops on Genotoxicity

Testing (IWGT). Mutation Research - Genetic Toxicology and Environmental Mutagenesis 847:

403022 (2019). doi: 10.1016/j.mrgentox.2019.02.003

Desmedt B., Binik S. Adam R., Rogiers V., ‘Baby care products’ in Handbook of Cosmetic

Science and Technology, Fourth Edition 4th Edition, edited by André O. Barel, Marc Paye

Howard I. Maibach, p.483-496 (2014).

Dey S, Carr G, Li, Brink, Zhou, Probabilistic Monte Carlo estimation for quantitative exposure

assessment of lotion transfer via baby wipes usage. Regul Toxicol Pharmacol. 79:54-63

(2016a). doi: 10.1016/j.yrtph.2016.05.006

Dey S., Purdon M., Kirsch T., Helbich HM., Kerr K., Lijuan L., Shaoying Z., Exposure Factor

considerations for safety evaluation of modern disposable diapers. Regul Toxicol Pharmacol

81:83-193 (2016b). doi: 10.1016/j.yrtph.2016.08.017

DG24/XXIV/1891/98: Mandate for the SCCNFP Specific Working Group on Inventory, 2

March 1998.

Diffey B.L. People do not apply enough sunscreen for protection. B.M.J. 313, 942 (1996).

doi: 10.1136/bmj.313.7062.942

DiMarco RL., Hunt DR., Dewi RE., Heilshorn SC., Improvement of paracellular transport in

the Caco-2 drug screening model using protein-engineered substrates, Biomaterials 129:152-

162 (2017). doi: 10.1016/j.biomaterials.2017.03.023

Doktorova T.Y., Pauwels M., Vinken M., Vanhaecke T., Rogiers V. Opportunities for an

alternative integrating testing strategy for carcinogen hazard assessment? Crit Rev Toxicol

42(2):91-106 (2012). doi: 10.3109/10408444.2011.623151

Dornic N., Ficheux A.C., Roudot AC., Consumption of cosmetic products by the French

population. Third part: Product exposure amount, Food Chem Toxicol 106: 209-222 (2017a).

doi: 10.1016/j.fct.2017.05.049

Dornic N., Ficheux A.C., Bernard A., Roudot AC., Adequacy of the default values for skin

surface area used for risk assessment and French anthropometric data by a probabilistic

approach, Food Chem Toxicol 106(pt A):386-392 (2017b). doi: 10.1016/j.fct.2017.06.017

_____________________________________________________________________________________________

110

Dornic N., Ficheux A.C., Roudot AC., Consumption of cosmetic products by the French

population. Second part, Food Chem Toxicol 106:130-141 (2017c). doi:

10.1016/j.fct.2016.02.008

Dudzina T., Delmaar CJ., Biesterbos JW, Bakker MI, Bokkers BG, Scheepers PT, van Engelen

JG, Hungerbuehler K, von Goetz N, The probabilistic aggregate consumer exposure model

(PACEM): validation and comparison to a lower-tier assessment for the cyclic siloxane D5.,

Environ Int. 79:8-16 (2015). doi: 10.1016/j.envint.2015.03.006

Dybing E., Sanner T., Roelfzema H., Kroese D. and Tennant R.W. T25: A simplified

carcinogenic potency index: Description of the system and study of correlations between

carcinogenic potency and species/site specificity and mutagenicity. Pharmacol Toxicol

80(6):272-279 (1997). doi: 10.1111/j.1600-0773.1997.tb01973.x

EC A.1 -Melting / freezing temperature Council Regulation (EC) No 440/2008 of 30 May 2008

laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European

Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction

of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.4.

EC A.2 - Boiling temperature Council Regulation (EC) No 440/2008 of 30 May 2008 laying

down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament

and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.14.

EC A.3 - Relative density Council Regulation (EC) No 440/2008 of 30 May 2008 laying down

test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of

the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.21.

EC A.4 - Vapour pressure Council Regulation (EC) No 440/2008 of 30 May 2008 laying down

test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of

the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.26.

EC A.6 - Water solubility Council Regulation (EC) No 440/2008 of 30 May 2008 laying down

test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of

the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.57.

EC A.8 - Partition coefficient Council Regulation (EC) No 440/2008 of 30 May 2008 laying

down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament

and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.67.

EC A.9 - Flash-point Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test

methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the

Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

Official Journal L 142, 31/05/2008, p.80.

EC A.15 - Auto-ignition temperature (liquids and gases) Council Regulation (EC) No 440/2008

of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.104.

EC B.1- Acute toxicity (oral) - Commission Directive 92/69/EEC of 31 July 1992 adapting to

technical progress for the seventeenth time Council Directive 67/548/EEC on the

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111

approximation of laws, regulations and administrative provisions relating to the classification,

packaging and labelling of dangerous substances. Official Journal L 383A, 29/12/1992 p.110.

EC B.1 bis - Acute oral toxicity - Fixed Dose Procedure Council Regulation (EC) No 440/2008

of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.145.

EC B.1 tris - Acute oral toxicity - Acute toxic class method Council Regulation (EC) No

440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No

1907/2006 of the European Parliament and of the Council on the Registration, Evaluation,

Authorisation and Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008,

p.158.

EC B.2 – Acute toxicity (inhalation) Council Regulation (EC) No 440/2008 of 30 May 2008

laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European

Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction

of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.174.

EC B.3 - Acute toxicity (dermal) Council Regulation (EC) No 440/2008 of 30 May 2008 laying

down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament

and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.178.

EC B.4 - Acute toxicity: dermal irritation / corrosion Council Regulation (EC) No 440/2008 of

30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.182.

EC B.5 - Acute toxicity: eye irritation / corrosion Council Regulation (EC) No 440/2008 of 30

May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.191.

EC B.6 - Skin sensitisation Council Regulation (EC) No 440/2008 of 30 May 2008 laying down

test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of

the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.202.

EC B.7 - Repeated dose (28 days) toxicity (oral) Council Regulation (EC) No 440/2008 of 30

May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.210.

EC B.8 - Repeated dose (28 days) toxicity (inhalation) Council Regulation (EC) No 440/2008

of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.216.

EC B.9 - Repeated dose (28 days) toxicity (dermal) Council Regulation (EC) No 440/2008 of

30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.221.

EC B.26 - Sub-chronic oral toxicity test: repeated dose 90-day oral toxicity study in rodents

Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to

Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the

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112

Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH). Official Journal

L 142, 31/05/2008, p.302.

EC B.27 - Sub-chronic oral toxicity test: repeated dose 90-day oral toxicity study in non-

rodents. Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods

pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on

the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH). Official

Journal L 142, 31/05/2008, p.308.

EC B.28 - Sub-chronic dermal toxicity study: 90-day repeated dermal dose study using

rodent species Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test

methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the

Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

Official Journal L 142, 31/05/2008, p.314.

EC B.29 - Sub-chronic inhalation toxicity study: 90-day repeated inhalation dose study using

rodent species Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test

methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the

Council on the Registration, Evaluation, Authorisation an Restriction of Chemicals (REACH).

Official Journal L 142, 31/05/2008, p.318.

EC B.30 - Chronic toxicity test Council Regulation (EC) No 440/2008 of 30 May 2008 laying

down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament

and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.323.

EC B.31 - Prenatal developmental toxicity study Council Regulation (EC) No 440/2008 of 30

May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.329.

EC B.33 - Combined chronic toxicity / carcinogenicity test Council Regulation (EC) No

440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No

1907/2006 of the European Parliament and of the Council on the Registration, Evaluation,

Authorisation and Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008,

p.344.

EC B.35 - Two-generation reproduction toxicity test Council Regulation (EC) No 440/2008 of

30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.355.

EC B.36 – Toxicokinetics Council Regulation (EC) No 440/2008 of 30 May 2008 laying down

test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of

the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 142, 31/05/2008, p.365.

EC B.40bis - In vitro skin corrosion: Human skin model test Council Regulation (EC) No

440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No

1907/2006 of the European Parliament and of the Council on the Registration, Evaluation,

Authorisation and Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008,

p.394.

EC B.41 - In vitro 3T3 NRU phototoxicity test Council Regulation (EC) No 440/2008 of 30

May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.400.

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EC B.42 - Skin sensitisation: Local Lymph Node Assay Commission Regulation (EU) No

640/2012 of 6 July 2012 amending, for the purpose of its adaptation to technical progress,

Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No

1907/2006 of the European Parliament and of the Council on the Registration, Evaluation,

Authorisation and Restriction of Chemicals (REACH). Official Journal L 193, 20/07/2012, p. 3.

EC B.42 - Skin sensitisation: Local Lymph Node Assay Council Regulation (EC) No 440/2008

of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.414. Amended by OJ

L193:

EC B.44 - Skin absorption: In vivo method Council Regulation (EC) No 440/2008 of 30 May

2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European

Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction

of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.432.

EC B.45 - Skin absorption: In vitro method Council Regulation (EC) No 440/2008 of 30 May

2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European

Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction

of Chemicals (REACH). Official Journal L 142, 31/05/2008, p.438.

EC B.46 - In vitro skin irritation: Reconstructed human epidermis model test Commission

Regulation (EC) No 761/2009 of 23 July 2009 amending, for the purpose of its adaptation to

technical progress, Regulation (EC) No 440/2008 laying down test methods pursuant to

Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the

Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH). Official Journal

L 220, 24/08/2009, p.24. Amended by OJ L193

EC B.46 – In vitro skin irritation: Reconstructed human epidermis test method Commission

Regulation (EU) No 640/2012 of 6 July 2012 amending, for the purpose of its adaptation to

technical progress, Regulation (EC) No 440/2008 laying down test methods pursuant to

Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the

Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH). Official Journal

L 193, 20/07/2012, p. 17.

EC B.47 – Bovine corneal opacity and permeability test method for identifying ocular

corrosives and severe irritants. Commission Regulation (EC) No 761/2009 of 23 July 2009

amending, for the purpose of its adaptation to technical progress, Regulation (EC) No

440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the

European Parliament and of the Council on the Registration, Evaluation, Authorisation and

Restriction of Chemicals (REACH). Official Journal L 324, 09/12/2010, p. 14.

EC B.48 – Isolated chicken eye test method for identifying ocular corrosives and severe

irritants. Commission Regulation (EC) No 761/2009 of 23 July 2009 amending, for the

purpose of its adaptation to technical progress, Regulation (EC) No 440/2008 laying down

test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of

the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals

(REACH). Official Journal L 324, 09/12/2010, p. 14.

EC B.50 – Skin sensitisation: Local Lymph Node Assay: DA Commission Regulation (EU) No

640/2012 of 6 July 2012 amending, for the purpose of its adaptation to technical progress,

Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No

1907/2006 of the European Parliament and of the Council on the Registration, Evaluation,

Authorisation and Restriction of Chemicals (REACH). Official Journal L 193, 20/07/2012, p.

46.

_____________________________________________________________________________________________

114

EC B.51 – Skin sensitisation: Local Lymph Node Assay: BrdU-ELISA Commission Regulation

(EU) No 640/2012 of 6 July 2012 amending, for the purpose of its adaptation to technical

progress, Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation

(EC) No 1907/2006 of the European Parliament and of the Council on the Registration,

Evaluation, Authorisation and Restriction of Chemicals (REACH). Official Journal L 193,

20/07/2012, p. 56.

EC B.52 - Acute Inhalation Toxicity: Acute Toxic Class Method: Commission Regulation (EU)

No 260/2014 of 24 January 2014 amending, for the purpose of its adaptation to technical

progress, Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation

(EC) No 1907/2006 of the European Parliament and of the Council on the Registration,

Evaluation, Authorisation and Restriction of Chemicals (REACH). Official Journal L 81,

19/03/2014 p. 1.

ECB (European Chemicals Bureau) Report from the Expert Working Group on Sensitisation.

Ispra, 4-6 November 2002. ECBI/81/02 Rev. 3., 2002.

ECB (European Chemicals Bureau) Technical Guidance Document on Risk Assessment in

support of Commission Directive 93/67/EEC on Risk Assessment for new notified substances,

Commission Regulation (EC) No 1488/94 on Risk Assessment for existing substances and

Directive 98/8/EC of the European Parliament and of the Council concerning the placing of

biocidal products on the market. Doc. EUR 20418 EN/1, European Communities (2003).

ECETOC Percutaneous absorption. European Centre for Ecotoxicology and Toxicology of

Chemicals (ECETOC), Monograph No 20, Brussels (1993).

ECETOC Recognition of, and Differentiation between, Adverse and Non-adverse Effects in

Toxicology Studies. European Centre for Ecotoxicology and Toxicology of Chemicals

(ECETOC), Technical Report No. 85, Brussels (2002).

ECHA (European Chemicals Agency) 2012a. Guidance on information requirements and

chemical safety assessment. Chapter R.8: Characterisation of dose [concentration]-response

for human health. Updated version 2.1, November 2012. Available at:

http://echa.europa.eu/documents/10162/13632/information_requirements_r8_en.pdf.

ECHA (European Chemicals Agency) 2012b. Guidance on information requirements and

chemical safety assessment. Chapter R.15: Consumer exposure estimation. Updated version

2.1, October 2012. Available at:

https://echa.europa.eu/documents/10162/13632/information_requirements_r15_en.pdf

(Consulted December 2020).

ECHA (European Chemicals Agency) 2014 Factsheet: Interface between REACH and

Cosmetics regulations, ECHA-14-FS-04-EN, ISBN:978-92-9244-819-6, DoI:10.2823/64527.

Available at:

https://echa.europa.eu/documents/10162/13628/reach_cosmetics_factsheet_en.pdf

(Consulted December 2020).

ECHA (European Chemicals Agency) 2015. Guidance on information requirements and

chemical safety assessment. Chapter R.7a: Endpoint specific guidance. Updated version 4.0,

July 2015. Available at:

http://echa.europa.eu/documents/10162/13632/information_requirements_r7a_en.pdf

(consulted December 2020).

ECHA (European Chemicals Agency) 2017. Chapter R.7a: Endpoint specific guidance Version

6.0-Guidance on Information Requirements and Chemical Safety Assessment Chapter R.7a:

Endpoint specific guidance. Version 6.0 (2017) doi:10.2823/337352

_____________________________________________________________________________________________

115

ECHA (European Chemical Agency) 2019a. Board of Appeal Decision, Case No. A-014-2018

- A-021-2018, 15 April 2019. Available at:

https://echa.europa.eu/documents/10162/cd6eedc4-98a6-c94f-618d-bd75878173ad

ECHA (European Chemicals Agency) 2019b. Appendix R.8-17 to Chapter R.8: Guidance for

preparing a scientific report for health based exposure limits at the workplace. Version 1.0 –

August 2019.

ECHA (European Chemical Agency) 2020a. Board of Appeal Decision, Case No. A-013-2018,

23 June 2020. Available at: https://echa.europa.eu/documents/10162/27c0fc88-360e-0321-

7b6a-e603ba0fd604.

ECHA (European Chemical Agency) 2020b. Board of Appeal Announcement, Case No. A-

002-2020, 16 June 2020. Available at:

https://echa.europa.eu/documents/10162/23010712/a-002-2020_announcement_en.pdf

EFSA 2009 (European Food Safety Authority) Use of the benchmark dose approach in risk

assessment. Guidance of the Scientific Committee. The EFSA Journal 1150, 1-72 (2009). doi:

10.2903/j.efsa.2009.1150

EFSA 2011a (European Food Safety Authority) Scientific opinion on genotoxicity testing

strategies applicable to food and feed safety assessment. The EFSA Journal 2379, 1-68

(2011a). doi: 10.2903/j.efsa.2011.2379

EFSA 2011b (European Food and Safety Authority) Guidance on the risk assessment of the

application of nanoscience and nanotechnologies in the food and feed chain, EFSA Journal

9(5): 2140, 36 pp, available at: www.efsa.europa.eu/en/efsajournal/pub/2140.htm,

(2011b). doi: 10.2903/j.efsa.2011.2140

EFSA 2012 (European Food Safety Authority) Scientific Opinion on Exploring options for

providing advice about possible human health risks based on the concept of Threshold of

Toxicological Concern (TTC). EFSA Journal;10(7):2750 (2012). doi:

10.2903/j.efsa.2012.2750

EFSA 2012a (European Food Safety Authority). Guidance on selected default values to be

used by the EFSA Scientific Committee, Scientific Panels and Units in the absence of actual

measured data. EFSA Journal 10(3):2579 (2012). doi:10.2903/j.efsa.2012.2579.

EFSA 2012b (European Food Safety Authority) Panel on Plant Protection Products and their

Residues (PPR). Guidance on Dermal Absorption (Scientific Opinion). EFSA Journal (2012).

doi: 10.2903/j.efsa.2012.2665

EFSA 2016 (European Food Safety Authority) Technical report on the outcome of the

pesticides peer review meeting on general recurring issues in mammalian toxicology. EFSA

supporting publication 2016: EN-1074. 24 pp., 2016.

EFSA 2016a (European Food Safety Authority) Review of the Threshold of Toxicological

Concern (TTC) approach and development of new TTC decision tree European Food Safety

Authority and World Health Organization EFSA Supporting Publications Volume 13, Issue 3

March 2016. doi.org/10.2903/sp.efsa.2016.EN-1006

EFSA 2017 (European Food Safety Authority). Guidance on dermal absorption. EFSA Journal

15(6):4873 (2017). doi: 10.2903/j.efsa.2017.4873

EFSA 2017a (European Food Safety Authority) Guidance on the use of the weight of

evidence approach in scientific assessments, EFSA Journal 15(8): 4971, August 2017. doi:

10.2903/j.efsa.2017.4971.

_____________________________________________________________________________________________

116

EFSA 2017b (European Food Safety Authority) EFSA Scientific Committee Scientific Opinion

on the clarification of some aspects related to genotoxicity assessment. EFSA Journal

2017;15(12):5113, 25 pp, 2017. doi: 10.2903/j.efsa.2017.5113

EFSA 2017c (European Food Safety Authority) EFSA Scientific Committee, Update: Guidance

on the use of the benchmark dose approach in risk assessment. EFSA Journal

2017;15(1):4658, 41 pp., 2017. doi: 10.2903/j.efsa.2017.4658

EFSA 2018 (European Food Safety Authority) Guidance on risk assessment of the application

of nanoscience and nanotechnologies in the food and feed chain: Part 1, human and animal

health. EFSA Journal; 16(7):5327 (2018). doi: 10.2903/j.efsa.2018.5327

EFSA 2019a (European Food Safety Authority) Guidance on the use of the Threshold of

Toxicological Concern approach in food safety assessment. EFSA Journal 17(6):5708 (2019).

doi: 10.2903/j.efsa.2019.5708

EFSA 2019b Fürst, P., Milana, MR., Pfaff, K., Tlustos, C., Vleminckx, C., Arcella, D.,

Barthélémy, E., Colombo, P., Goumperis, T., Roldán Torres, R., Pasinato. and Afonso, A.

Scientific technical assistance to RASFF on chemical contaminants: Risk evaluation of

chemical contaminants in food in the context of RASFF notifications. EFSA supporting

publication 2019:EN-1625. 108 pp. doi:10.2903/sp.efsa.2019.EN-1625

EFSA, ECHA, JRC Guidance for the identification of endocrine disruptors in the context of

Regulations (EU) No 528/2012 and (EC) No 1107/2009,. EFSA Journal;16(6):5311 (2018).

doi: 10.2903/j.efsa.2018.5311

Ellison, C. A., Blackburn, K. L., Carmichael, P. L., Clewell, H. J., Cronin, M. T. D., Desprez,

B., Escher, S. E., Ferguson, S. S., Grégoire, S., Hewitt, N. J., Hollnagel, H. M., Klaric, M.,

Patel, A., Salhi, S., Schepky, A., Schmitt, B. G., Wambaugh, J. F. & Worth, A. Challenges in

working towards an internal threshold of toxicological concern (iTTC) for use in the safety

assessment of cosmetics: Discussions from the Cosmetics Europe iTTC Working Group

workshop. Regulatory Toxicology and Pharmacology 103: 63–72 (2019). doi:

10.1016/j.yrtph.2019.01.016

EMA 2012 ICH Guideline S10. Guidance on photosafety testing of pharmaceuticals. European

Medicines Agency EMA/CHMP/ICH/752211 (2012).

EMA 2015 Committee for Human Medicinal Products, ICH Guidance S10 on Photosafety

Evaluation of Pharmaceuticals, Step 5, EMA/CHMP/ICH/752211/2012, 25 August 2015.

ESAC 2001 Statement on the scientific validity of the Embryonic Stem Cell Test (EST), the

Micromass Test and the Postimplantation Rat Whole-Embryo Culture Assay – in vitro tests

for embryotoxicity. Adopted by the ECVAM Scientific Advisory Committee (ESAC) at its 17

Meeting of 16-17 October 2001 at ECVAM, Ispra, Italy (2001).

Escher, S. E., Tluczkiewicz, I., Batke, M., Bitsch, A., Melber, C., Kroese, E. D., Buist, H. E. &

Mangelsdorf, I. Evaluation of inhalation TTC values with the database RepDose. Regul Toxicol

Pharmacol 58(2): 259–274 (2010). doi: 10.1016/j.yrtph.2010.06.009

Escher, S. E., Mangelsdorf, I., Hoffmann-Doerr, S., Partosch, F., Karwath, A., Schroeder, K.,

Zapf, A. & Batke, M. Time extrapolation in regulatory risk assessment: The impact of study

differences on the extrapolation factors. Regul Toxicol Pharmacol 112: 104584 (2020). doi:

doi: 10.1016/j.yrtph.2020.104584

Eskes A. and Zuang V. Alternative (Non-Animal) Methods for Cosmetic Testing: Current

Status and Future Prospects. ATLA, 33 (suppl. 1):1-227 (2005).

_____________________________________________________________________________________________

117

EUR 10305: Reports of the Scientific Committee on Cosmetology. Fourth series. 1986.

EUR 11080: Reports of the Scientific Committee on Cosmetology. Fifth series. 1987.

EUR 11139: Reports of the Scientific Committee on Cosmetology. Sixth series. 1987.

EUR 11303: Reports of the Scientific Committee on Cosmetology. Seventh series. 1988.

EUR 14208: Opinion of the Scientific Committee on Cosmetology. (11/86 — 10/90). 1993.

EUR 7297: Reports of the Scientific Committee on Cosmetology. First series. 1982.

EUR 8634: Reports of the Scientific Committee on Cosmetology. Second series. 1983.

EUR 8794: Reports of the Scientific Committee on Cosmetology. Third series. 1983.

EURL ECVAM. Recommendation concerning the cell transformation assays (CTA) using

Syrian Hamster Embryo cells (SHE) and the BALB/c 3T3 mouse fibroblast cell line for in vitro

carcinogenicity testing, including the ESAC opinion (Annex 1) based on the ESAC peer review

of an EURL ECVAM-coordinated validation study of three CTA protocols for in vitro

carcinogenicity testing (2012) Available at: https://ihcp.jrc.ec.europa.eu/our_labs/eurl-

ecvam/eurl-ecvamrecommendations/cta-recommendation

EURL ECVAM. JRC (Joint Research Centre) Scientific and Policy Reports: EURL ECVAM

Recommendation on the 3T3 Neutral Red Uptake Cytotoxicity Assay for Acute Oral Toxicity

Testing. (April 2013). JRC79556, EUR 25946 EN. (2013). doi: 10.2788/8883 (print)

10.2788/88799 (online)

Evans, S. J., Clift, M. J. D., Singh, N., De Oliveira Mallia, J., Burgum, M., Wills, J. W.,

Wilkinson, T. S., Jenkins, G. J. S. & Doak, S. H. Critical review of the current and future

challenges associated with advanced in vitro systems towards the study of nanoparticle

(secondary) genotoxicity. Mutagenesis 32: 233–241 (2017). doi: 10.1093/mutage/gew054

Evans, S. J., Gollapudi, B., Moore, M. M. & Doak, S. H. Horizon scanning for novel and

emerging in vitro mammalian cell mutagenicity test systems. Mutat Res Gen Tox En 847:

403024 (2019a). doi: 10.1016/j.mrgentox.2019.02.005

Evans, S. J., Clift, M. J. D., Singh, N., Wills, J. W., Hondow, N., Wilkinson, T. S., Burgum, M.

J., Brown, A. P., Jenkins, G. J. & Doak, S. H. In vitro detection of in vitro secondary

mechanisms of genotoxicity induced by engineered nanomaterials. Part Fibre Toxicol 16(1):

(2019b). doi: 10.1186/s12989-019-0291-7

Ezendam J., Braakhuis H. M. and Vandebriel R. J., State of the art in non-animal approaches

for skin sensitization testing: from individual test methods towards testing strategies. Arch

Toxicol 90(12), 2861-2883, (2016). doi: 10.1007/s00204-016-1842-4

Fang Y., Eglen R.M. Three-Dimensional Cell Cultures in Drug Discovery and Development.

SLAS Discov 22(5):456–472 (2017). doi: 10.1177/1087057117696795

Fasano W.J., Manning L.A., Green J.W. Rapid integrity assessment of rat and human

epidermal membranes for in vitro dermal regulatory testing: correlation of electrical

resistance with tritiated water permeability. Toxicol In vitro 16(6):731–740 (2002). doi:

10.1016/s0887-2333(02)00084-x

FDA 2015 S10 Photosafety Evaluation of Pharmaceuticals. Guidance for Industry U.S.

Department of Health and Human Services, Food and Drug Administration (2015).

 
 
 
 
 
_____________________________________________________________________________________________ 
118 
 
Feigenbaum,  A.,  Pinalli,  R., Giannetto,  M.  &  Barlow,  S.  Reliability  of the  TTC  approach: 
Learning from inclusion of pesticide active substances in the supporting database. Food Chem 
Toxicol 75: 24–38 (2015). doi: 10.1016/j.fct.2014.10.016 
Felter S.P, Daston GP., Euling SY., Piersma AH., Tassinari, MS. Assessment of health risks 
resulting from early-life exposures. Are current chemical toxicity testing protocols and risk 
assessment  methods  adeaquate?  Crit  Rev  Toxicol,  45(3):219-244  (2015).  doi:  doi: 
10.3109/10408444.2014.993919. 
 
Felter SP., Carr AN., Zhu T., Kirsch T., Niu G. Safety evaluation for ingredients used in baby 
care products:  Consideration of diaper rash.  Regul Toxicol Pharmacol 90:214-221  (2017). 
doi: 10.1016/j.yrtph.2017.09.011 
 
Ficheux AS., Dornic N., Bernard A., Chevillotte G., Roudot AC. Probabilistic assessment of 
exposure to cosmetic products by French children aged 0-3 years, Food Chem Toxicol 94:85-
92 (2016a). doi: 10.1016/j.fct.2016.05.02 
 
Ficheux AS., Bernard A., Chevillotte G., Dornic N., Roudot AC., Probabilistic assessment of 
exposure to hair cosmetic products by the French population, Food Chem Toxicol 92:205-216 
(2016b). doi: 10.1016/j.fct.2016.04.009 
 
Ficheux  AS.,  Chevillotte  G.,  Wesolek  N.,  Morisset  T.,  Dornic  N.,  Bernard  A.,  Bertho  A., 
Romanet A., Leroy L., Mercat AC., Creusot T., Simon E., Roudot AC., Consumption of cosmetic 
products by the French population second part: Amount data. Food Chem Toxicol 90:130-
141 (2016c). doi: 10.1016/j.fct.2016.02.008 
 
Ficheux AS. and Roudot AC. Exposition de la population Francaise aux produits cosmétiques, 
Université  Bretagne,  Loire  and  université  de  Bretagne  Occidentale,  p1-p670  (2017)  - 
COSMED. 
 
Ficheux, AS., Gomez-Berrada, M. P., Roudot, A. C. & Ferret, P. J. Consumption and exposure 
to finished cosmetic products: A systematic review. Food and Chemical Toxicology vol. 124 
280–299 (2019). doi: 10.1016/j.fct.2018.11.060 
Fluhr, J. W. & Darlenski, R. Skin Surface pH in Newborns: Origin and Consequences. Curr 
Probl Dermatol. 54: 26–32 (2018). 
Galli  CL.,  Sensi  C.,  Fumagalli  A.,  Parravicini  C.,  Marinovich  M.,  Eberini  I.  Computational 
Approach to Evaluate the Androgenic Affinity of Iprodione, Procymidone, Vinclozolin and their 
metabolites, PLOS ONE 9(8): e104822 (2014). doi: 10.1371/journal.pone.0104822 
 
Garcia-Hidalgo E., von Goetz N., Siegrist  M., Hungerbühler K., Use-patterns of personal 
care and household cleaning products in Switzerland. Food Chem Toxicol 99:24-39 (2017). 
doi: 10.1016/j.fct.2016.10.030 
 
Gerstel D., Jacques-Jamin C., Schepky A., Cubberley R., Eilstein J., Grégoire S., Hewitt N., 
Klaric M., Rothe H., Duplan H. Comparison of protocols for measuring cosmetic ingredient 
distribution  in  human  and  pig  skin.  Toxicol  In  vitro  34:153-160  (2016).  doi: 
10.1016/j.tiv.2016.03.012 
Gilmour, N., Kern, P. S., Alépée, N., Boislève, F., Bury, D., Clouet, E., Hirota, M., Hoffmann, 
S., Kühnl, J., Lalko, J. F., Mewes, K., Miyazawa, M., Nishida, H., Osmani, A., Petersohn, D., 
Sekine,  S.,  van  Vliet,  E.  &  Klaric,  M.  Development  of  a  next  generation  risk  assessment 
framework  for  the  evaluation  of  skin  sensitisation  of  cosmetic  ingredients.  Regul  Toxicol 
Pharmacol 116: 104721 (2020). doi: 10.1016/j.yrtph.2020.104721 

_____________________________________________________________________________________________

119

Gocke E., Albertini S., Chetelat A., Kirchner S., Muster W. The photomutagenicity of

fluoroquinolones and other dyes. Toxicol Lett 102/103:375-381 (1998). doi: 10.1016/s0378-

4274(98)00235-5

Goebel, C., Troutman, J., Hennen, J., Rothe, H., Schlatter, H., Gerberick, G. F. & Blömeke,

B. Introduction of a methoxymethyl side chain into p-phenylenediamine attenuates its

sensitizing potency and reduces the risk of allergy induction. Toxicol Appl Pharmacol 274 (3):

480-487 (2014). doi:10.1016/j.taap.2013.11.016.

Gold L.S., Sawyer C.B., Magaw R., Backman G.M., Deveciana M., Levinson R., Hooper N.K.,

Havender W.R, Bernstein L., Peto R., Pike M.C., Ames B.N. A Carcinogenic Potency Database

of the Standardized Results of Animal Bioassays. Environ Health Pers 58:9-319 (1984). doi:

10.1289/ehp.84589

Gomez-Berrada MP., Ficheux AS., Boudières I., Chiter M., Rielland A., De Javel D., Roudot

AC., Ferret PJ. Consumption and exposure assessment to toothpaste in French families. Food

Chem Toxicol 118:24-31 (2018a). doi: 10.1016/j.fct.2018.04.061

Gomez-Berrada MP., Ficheux AS., Rakotomalala S., Guillou S., Belle, De Javel D.,

RoudotAC., Ferret PJ. Consumption and exposure assessment to sunscreen products: A key

point for safety assessment. Food Chem Toxicol 114:170-179 (2018b). doi:

10.1016/j.fct.2018.02.035

Gomez-Berrada MP., Ficheux S., Guillou S., Berge C., de Javel D., Roudot AC., Ferreta PJ.,

Consumption and exposure assessment to cosmetic products for children under 2 years old.

Food Chem Toxicol 105:151-160 (2017). doi: 10.1016/j.fct.2017.04.011

Goodman, J. I. Erratum: Goodbye to the bioassay. Toxicol Res 7(5): 994 (2018). doi:

10.1039/c8tx90016g Goodsaid F.M., Amur S., Aubrecht J., Burczynski M.E., Carl K.,

Catalano J., Charlab R., Close S., Cornu-Artis C., Essioux L., Fornace AJ Jr., Hinman L., Hong

H., Hunt I., Jacobson-Kram D., Jawaid A., Laurie D., Lesko L., Li H.H., Lindpaintner K., Mayne

J., Morrow P., Papaluca-Amati M., Robison T.W., Roth J., Schuppe-Koistinen I., Shi L., Spleiss

O., Tong W., Truter S.L., Vonderscher J., Westelinck A., Zhang L., Zineh I. Voluntary

exploratory data submissions to the US FDA and the EMA: experience and impact. Nature

Reviews. Drug discovery 9(6):435-445 (2010). doi: 10.1038/nrd3116

Gottlieb A., Bourget T.D., Lowe N.J. Sunscreens: effects of amounts of application of sun

protection factors. In: Lowe NJ, Shaat NA, Pathak MA eds. Sunscreens: development,

evaluation, and regulatory aspects. New York, Marcel Dekker, pp. 583-588 (1997).

Grès M-C., Julian B., Bourrié M., Meunier V., Roques C., Berger M., Boulenc X., Berger Y.,

Fabre G. Correlation between oral drug absorption in humans and apparent drug permeability

in TC-7 cells, a human epithelial intestinal cell line: comparison with the parental Caco-2 cell

line. Pharm Res 15(5):726-733 (1998). doi: 10.1023/a:1011919003030

Gundert-Remy U., Bernauer U., Blömeke B., Döring B., Fabian E., Goebel C., Hessel S.,

Jäckh C., Lampen A., Oesch F., Petzinger E., Völkel W., Roos P.H. Extrahepatic metabolism

at the body's internal-external interfaces. Drug Metab Rev. 46(3):291-324 (2014). doi:

10.3109/03602532.2014.900565

Gustin, J., Bohman, L., Ogle, J., Chaudhary, T., Li, L., Fadayel, G., Mitchell, M. C., Narendran,

V., Visscher, M. O. & Carr, A. N. Use of an emollient‐containing diaper and pH‐buffered wipe

regimen restores skin pH and reduces residual enzymatic activity. Pediatr Dermatol 37: 626–

631 (2020). doi: 10.1111/pde.14169

Guth K., Schäfer-Korting M., Fabian E., Landsiedel R., van Ravenzwaay B., Suitability of skin

integrity tests for dermal absorption studies in vitro. Toxicol in vitro 29(1):113–123 (2015).

doi: 10.1016/j.tiv.2014.09.007

 
 
 
 
 
_____________________________________________________________________________________________ 
120 
 
 
Hall B., Steiling W., Safford B., Coroama M., Tozer S., Firmani C., McNamara C., Gibney M. 
European consumer exposure to cosmetic products, a framework for conducting population 
exposure  assessments,  Part  2.  Food  Chem  Toxicol  49(2):408-22  (2011).  doi: 
10.1016/j.fct.2010.11.016 
 
Hall B., Tozer S., Safford B., Coroama M., Steiling W., Leneveu-Duchemin M.C., McNamara 
C., Gibney M. European consumer exposure to cosmetic products, a framework for conducting 
population  exposure  assessments.  Food  Chem  Toxicol  45(11):2097-2108  (2007).  doi: 
10.1016/j.fct.2007.06.017 
Hasselgren, C., Ahlberg, E., Akahori, Y., Amberg, A., Anger, L. T., Atienzar, F., Auerbach, 
S., Beilke, L., Bellion, P., Benigni, R., Bercu, J., Booth, E. D., Bower, D., Brigo, A., Cammerer, 
Z., Cronin, M. T. D., Crooks, I., Cross, K. P., Custer, L., Dobo, K., Doktorova, T., Faulkner, 
D., Ford,  K. A.,  Fortin, M.  C., Frericks,  M., Gad-McDonald, S.  E., Gellatly, N.,  Gerets, H., 
Gervais, V., Glowienke, S., Van Gompel, J., Harvey, J. S., Hillegass, J., Honma, M., Hsieh, J. 
H., Hsu, C. W., Barton-Maclaren, T. S., Johnson, C., Jolly, R., Jones, D., Kemper, R., Kenyon, 
M.  O.,  Kruhlak,  N.  L.,  Kulkarni,  S.  A.,  Kümmerer,  K.,  Leavitt,  P.,  Masten,  S.,  Miller,  S., 
Moudgal, C., Muster, W., Paulino, A., Lo Piparo, E., Powley, M., Quigley, D. P., Reddy, M. V., 
Richarz, A. N., Schilter, B., Snyder, R. D., Stavitskaya, L., Stidl, R., Szabo, D. T., Teasdale, 
A., Tice, R. R., Trejo-Martin, A., Vuorinen, A., Wall, B. A., Watts, P., White, A. T., Wichard, 
J., Witt, K. L., Woolley, A., Woolley, D., Zwickl, C. & Myatt, G. J. Genetic toxicology in silico 
protocol. Regul Toxicol Pharmacol 107: 104403 (2019). doi: 10.1016/j.yrtph.2019.104403 
Hatherell, S., Baltazar, M. T., Reynolds, J., Carmichael, P. L., Dent, M., Li, H., Ryder, S., 
White, A., Walker, P. & Middleton, A. M. Identifying and characterizing stress pathways of 
concern for consumer safety in next-generation risk assessment. Toxicol Sci 176(1): 11–33 
(2020). doi:10.1093/toxsci/kfaa054. 
Health  Canada,  Science  approach  document  for  the  TTC-based  approach  for  certain 
substances, Canada Gazette, Part 1: vol 150, n° 40, October 1 (2016). 
 
Hewitt N.J., Edwards RJ., Fritsche E., Goebel C., Aeby P., Scheel J., Reisinger K., Ouédraogo 
G., Duche D., Eilstein J., Latil A., Kenny J., Moore C., Kuehnl J., Barroso J., Fautz R., Pfuhler 
S., Use of human in vitro skin models for accurate and ethical risk assessment: metabolic 
considerations. Toxicol Sci. 2013 133(2):209-17 (2013). doi: 10.1093/toxsci/kft080 
 
Hoffmann S, Kleinstreuer N., Alépée N., Allen D., Api AM., Ashikaga T., Clouet E., Cluzel M., 
Desprez B., Gellatly N., Goebel C., Kern PS., Klaric M., Kühnl J., Lalko JF., Martinozzi-Teissier 
S., Mewes  K., Miyazawa M., Parakhia  R., van Vliet E., Zang Q.,  Petersohn  D. Non-animal 
methods to predict skin sensitization (I): the Cosmetics Europe database. Crit Rev Toxicol. 
48(5):344-358 (2018). doi: 10.1080/10408444.2018.1429385 
Honma, M., Kitazawa, A., Cayley, A., Williams, R. V, Barber, C., Hanser, T., Saiakhov, R., 
Chakravarti, S., Myatt, G. J., Cross, K. P., Benfenati, E., Raitano, G., Mekenyan, O., Petkov, 
P.,  Bossa,  C.,  Benigni,  R.,  Battistelli,  C.  L.,  Giuliani,  A.,  Tcheremenskaia,  O.,  DeMeo,  C., 
Norinder, U., Koga, H., Jose, C., Jeliazkova, N., Kochev, N., Paskaleva, V., Yang, C., Daga, 
P. R., Clark, R. D. & Rathman, J. Improvement of quantitative structure–activity relationship 
(QSAR) tools for predicting Ames mutagenicity: outcomes of the Ames/QSAR International 
Challenge Project. Mutagenesis 34: 3–16 (2019). doi: 10.1093/mutage/gey031 
Husøy T, Martínez MA, Sharma RP, Kumar V, Andreassen M, Sakhi AK, Thomsen C, Dirven 
H. Comparison of aggregated exposure to di(2-ethylhexyl) phthalate from diet and personal 
care products with urinary concentrations of metabolites using a PBPK model - Results from 
the Norwegian biomonitoring study in EuroMix. Food Chem Toxicol. (2020) Sep;143:111510. 
doi: 10.1016/j.fct.2020.111510. Epub 2020 Jun 30. PMID: 32615240. 

_____________________________________________________________________________________________

121

Ibrahim, M. A., Yasui, M., Saha, L. K., Sasanuma, H., Honma, M. & Takeda, S. Enhancing

the sensitivity of the thymidine kinase assay by using DNA repair-deficient human TK6 cells.

Environ Mol Mutagen 61: 602–610 (2020). doi: 10.1002/em.22371

ICCR In silico Approaches for Safety Assessment of Cosmetic Ingredients, A report for the

International Cooperation on Cosmetics Regulation (2014).

ICR publication 66 Human respiratory tract model for radiological prorection. A report of a

Task Group of the International Commission on Radiological Protection. Ann. ICRP 24(1-3):1-

482 (1994).

IP/06/1047: European Commission Press Release Commission bans 22 hair dye substances

to increase consumer safety, Brussels, 20 July 2006, available through

http://europa.eu/rapid/press-release_IP-06-1047_en.htm (Consulted December 2020).

Jacobs MN, Colacci A., Louekari K., Luijten M., Hakkert BC., Paparella M., Vasseur P.

International regulatory needs for development of an IATA for non-genotoxic carcinogenic

chemical substances. ALTEX. 33(4):359-392 (2016). doi: 10.14573/altex.1601201

Jacobs, M. N., Colacci, A., Corvi, R., Vaccari, M., Aguila, M. C., Corvaro, M., Delrue, N.,

Desaulniers, D., Ertych, N., Jacobs, A., Luijten, M., Madia, F., Nishikawa, A., Ogawa, K.,

Ohmori, K., Paparella, M., Sharma, A. K. & Vasseur, P. Chemical carcinogen safety testing:

OECD expert group international consensus on the development of an integrated approach

for the testing and assessment of chemical non-genotoxic carcinogens. Arch Toxicol 94(8):

2899–2923 (2020). doi: 10.1007/s00204-020-02784-5

Jacobs. M. N., In silico tools to aid risk assessment of endocrine disrupting chemicals. Toxicol

205(1-2):43-5 (2004). doi: 10.1016/j.tox.2004.06.036

Johansson H, Rydnert F, Kühnl J, Schepky A, Borrebaeck C, Lindstedt M, Genomic allergen

rapid detection in-house validation--a proof of concept, Toxicol Sci 139(2):362-70 (2014).

doi: 10.1093/toxsci/kfu046

Johansson H., Lindstedt M., Albrekt AS., Borrebaeck C. A genomic biomarker signature can

predict skin sensitizers using a cell-based in vitro alternative to animal tests. BMC Genomics,

8(12):399 (2011). doi: 10.1186/1471-2164-12-399

Johnson TN. The development of drug metabolising enzymes and their influence on the

susceptibility to adverse drug reactions in children. Toxicol 192(1):37-48 (2003). doi: doi:

10.1016/s0300-483x(03)00249-x

JRC (Joint Research Centre) Scientific and Policy Reports: EURL ECVAM Recommendation on

the 3T3 Neutral Red Uptake Cytotoxicity Assay for Acute Oral Toxicity Testing, JRC79556,

EUR 25946 EN (2013). doi:10.2788/88799.

JRC (Joint Research Centre) Scientific and Policy Reports: EURL ECVAM progress report on

validation and regulatory acceptance of alternative methods (2010-2013) prepared in the

framework of Directive 76/768/EEC and Regulation (EC) N° 1223/2009 on cosmetic products.

Valérie Zuang, Michael Schäffler et al. JRC 80506, EUR 25981 EN, 64 pp (2013a).

JRC (Joint Research Centre) Scientific and Policy Reports: EURL ECVAM status report on the

development, validation and regulatory acceptance of alternative methods and approaches

(2013-April 2014). JRC 90989, EUR 26702 EN, 84 pp (2014a).

JRC (Joint Research Centre) Scientific and Policy Reports: Alternative methods for regulatory

toxicology – a state-of-the-art review (Worth A et al.; Eds.), Report (2014b).

_____________________________________________________________________________________________

122

JRC (Joint Research Centre) Scientific and Policy Reports: EURL ECVAM strategy for achieving

3Rs impact in the assessment of toxicokinetics and systemic toxicity Bessems J., Coecke S.,

Gouliarmou V., Whelan M., Worth A., JRC96418, EUR 27315 EN, ISBN 978-92-79-49070-5

(2015).

JRC (Joint Research Centre) Zuang V., Dura A., et al., EURL ECVAM Status Report on the

Development, Validation and Regulatory Acceptance of Alternative Methods and Approaches,

EUR 28823 EN, Publications Office of the European Union, Luxembourg, 2016, ISBN 978-92-

79-63030-9 (print), doi:10.2787/644905, JRC 103522 (2016).

JRC (Joint Research Centre) Zuang V., Dura A., et al., EURL ECVAM Status Report on the

Development, Validation and Regulatory Acceptance of Alternative Methods and Approaches,

EUR 28823 EN, Publications Office of the European Union, Luxembourg, 2017, ISBN 978-92-

79-74268-2 (print), doi:10.2760/654653, JRC 108831 (2017).

JRC (Joint Research Centre) Zuang V., Dura A., et al., EURL ECVAM Status Report on the

Development, Validation and Regulatory Acceptance of Alternative Methods and Approaches,

EUR 29455 EN, Publications Office of the European Union, Luxembourg, 2018, ISBN 978-92-

79-97418-2 (print), doi:10.2760/858311, JRC113594 (2018).

JRC (Joint Research Centre) Zuang V., Dura A., et al., EURL ECVAM Status Report on the

Development, Validation and Regulatory Acceptance of Alternative Methods and Approaches,

EUR 30100 EN, Publications Office of the European Union, Luxembourg, 2020, ISBN 978-92-

76-16368-8, doi:10.2760/25602, JRC119292 (2019).

JRC (Joint Research Centre) Zuang V., Dura A., et al., EURL ECVAM Status Report 2020:

Non-animal Methods in Science and Regulation, EUR 30553 EN, Publications Office of the

European Union, Luxembourg, 2021, ISBN 978-92-76-28395-9, doi:10.2760/7 19755, JRC

123531(2020).

Kalkhof, H., Herzler, M., Stahlmann, R. & Gundert-Remy, U. Threshold of toxicological

concern values for non-genotoxic effects in industrial chemicals: Re-evaluation of the Cramer

classification. Arch Toxicol 86(1): 17–25 (2012). doi: 10.1007/s00204-011-0732-z

Kandarova H. Phototoxicity studies using reconstructed human tissue models, in ICH

Steering Committee and Expert Working Group Meetings, EWG S-10 – Photo Safety

Evaluation of Pharmaceuticals, Sevilla, Spain, November 5-10 (2011).

Karlberg A.T., Bergström M.A., Börje A., Luthman K., Nilsson J.L. Allergic contact dermatitis-

formation, structural requirements, and reactivity of skin sensitizers. Chem Res Toxicol

21(1):53–69 (2008). doi: 10.1021/tx7002239

Kawamoto, T., Fuchs, A., Fautz, R. & Morita, O. Threshold of Toxicological Concern (TTC)

for Botanical Extracts (Botanical-TTC) derived from a meta-analysis of repeated-dose toxicity

studies. Toxicol Lett 316: 1–9 (2019). doi: 10.1016/j.toxlet.2019.08.006

Kazem S., Linssen EC., Gibbs S. Skin metabolism phase I and phase II enzymes in native

and reconstructed human skin: a short review. Drug Discov Today. 24(9):1899-1910 (2019).

doi: 10.1016/j.drudis.2019.06.002.

Kersten B., Kasper P., Brendler-Schwaab S.Y., Muller L. Use of the photo-micronucleus assay

in Chinese hamster V79 cells to study photochemical genotoxicity. Mutat Res 519 (1-2):49-

66 (2002). doi: 10.1016/s1383-5718(02)00113-4.

Kirkland D, Aardema M, Henderson L, Muller L Evaluation of the ability of a battery of three

in vitro genotoxicity tests to discriminate rodent carcinogens and non-carcinogens I.

Sensitivity, specificity and relative predictivity. Mutat Res 584(1-2):1–256 (2005). doi:

10.1016/j.mrgentox.2005.02.004.

_____________________________________________________________________________________________

123

Kirkland D., Reeve L., Gatehouse D., Vanparys P. A core in vitro genotoxicity battery

comprising the Ames test plus the in vitro micronucleus test is sufficient to detect rodent

carcinogens and in vivo genotoxins. Mutat Res 721(1):27-73 (2011). doi:

10.1016/j.mrgentox.2010.12.015

Kirkland, D., Uno, Y., Luijten, M., Beevers, C., van Benthem, J., Burlinson, B., Dertinger, S.,

Douglas, G. R., Hamada, S., Horibata, K., Lovell, D. P., Manjanatha, M., Martus, H. J., Mei,

N., Morita, T., Ohyama, W. & Williams, A. In vivo genotoxicity testing strategies: Report from

the 7th International workshop on genotoxicity testing (IWGT). Mutat Res - Genet Toxicol

Environ Mutagen 847: 403035 (2019). doi: 10.1016/j.mrgentox.2019.03.008

Kleinstreuer NC, Hoffmann S., Alépée N., Allen D., Ashikaga T., Casey W., Clouet E., Cluzel

M., Desprez B., Gellatly N., Göbel C., Kern PS., Klaric M., Kühnl J., Martinozzi-Teissier S.,

Mewes K., Miyazawa M., Strickland J., van Vliet E., Zang Q., Petersohn D. Non-animal

methods to predict skin sensitization (II): an assessment of defined approaches. Crit Rev

Toxicol, 48(5):359-374 (2018). doi: 10.1080/10408444.2018.1429386.

Kopp, B., Khoury, L. & Audebert, M. Validation of the γH2AX biomarker for genotoxicity

assessment: a review. Arch Toxicol 93(8): 2103–2114 (2019). doi: 10.1007/s00204-019-

02511-9

Kortenkamp A, Martin O, Faust M, Evans R, McKinlay R, Orton F, Rosivatz E. (2011) State

of the art assessment of endocrine disrupters, Final Report Project Contract Number

070307/2009/550687/SER/D3.

http://ec.europa.eu/environment/chemicals/endocrine/pdf/sota_edc_final_report.pdf.

Kroes R, Renwick AG., Feron V., Galli CL., Gibney M., Greim H., Guy RH., Lhuguenot JC.,

van de Sandt JJ. Application of the threshold of toxicological concern (TTC) to the safety

evaluation of cosmetic ingredients. Food Chem Toxicol 45(12): 2533-2562 (2007). doi:

10.1016/j.fct.2007.06.021

La Merrill, M.A., Vandenberg, L.N., Smith, M.T. et al. Consensus on the key characteristics

of endocrine-disrupting chemicals as a basis for hazard identification. Nat Rev Endocrinol 16,

45–57 (2020). doi: 10.1038/s41574-019-0273-8

Laufersweiler, M. C., Gadagbui, B., Baskerville-Abraham, I. M., Maier, A., Willis, A., Scialli,

A. R., Carr, G. J., Felter, S. P., Blackburn, K. & Daston, G. Correlation of chemical structure

with reproductive and developmental toxicity as it relates to the use of the threshold of

toxicological concern. Regul Toxicol Pharmacol 62(1): 160–182 (2012). doi:

10.1016/j.yrtph.2011.09.004.

Le Ferrec E., Chesne C., Artusson P., Brayden D., Fabre G., Gires P., Guillou F., Rousset M.,

Rubas W., Scarino M-L. In vitro models of the intestinal barrier. The report and

recommendations of ECVAM workshop 46. ATLA 29:649-668 (2001). doi:

10.1177/026119290102900604.

Lee E., Yun J., Ha J., Park BC., Kim HR., Hong SP. Kim KB, Kim MH., Assessment of exposure

for baby cosmetic care products in a Korean population. Food Chem Toxicol 106:107-113

(2017). doi: 10.1016/j.fct.2017.05.039

Lehman et al. The Tritiated Water Skin Barrier Integrity Test: Considerations for Acceptance

Criteria with and Without 14C-Octanol. Pharm Res 34(1):217-228 (2017). doi:

10.1007/s11095-016-2057-3

Lelièvre D, Justine P, Christiaens F, Bonaventure N, Coutet J, Marrot L, Cotovio J. The Episkin

phototoxicity assay (EPA): Development of an in vitro tiered strategy using 17 reference

_____________________________________________________________________________________________

124

chemicals to predict phototoxic potency. Toxicol In vitro 21(6):977–995 (2007). doi:

10.1016/j.tiv.2007.04.012

Lemper M., De Paepe K., Adam R., Rogiers V. Baby care products. In: Barel A.O., Paye M.,

Maibach H.I., eds. Handbook of Cosmetic Science and Technology, 3

ed., New York, London:

Informa Healthcare:613-623 (2009).

Levy, D. D., Hakura, A., Elespuru, R. K., Escobar, P. A., Kato, M., Lott, J., Moore, M. M. &

Sugiyama, K. ichi. Demonstrating laboratory proficiency in bacterial mutagenicity assays for

regulatory submission. Mutat Res - Genet Toxicol Environ Mutagen 848: 403075 (2019a).

doi: 1 0.1016/j.mrgentox.2019.07.005

Levy, D. D., Zeiger, E., Escobar, P. A., Hakura, A., van der Leede, B. jan M., Kato, M., Moore,

M. M. & Sugiyama, K. ichi. Recommended criteria for the evaluation of bacterial mutagenicity

data (Ames test). Mutat Res - Genet Toxicol Environ Mutagen 848: 403074 (2019b). doi:

/10.1016/j.mrgentox.2019.07.004

Luijten, M., Ball, N. S., Dearfield, K. L., Gollapudi, B. B., Johnson, G. E., Madia, F., Peel, L.,

Pfuhler, S., Settivari, R. S., Burg, W., White, P. A. & Benthem, J. Utility of a next generation

framework for assessment of genomic damage: A case study using the industrial chemical

benzene. Environ Mol Mutagen 61: 94–113 (2020). doi : 10.1002/em.22346

Lynch I., Dawson K.A. Protein-nanoparticle interactions. Nano Today 3:40-7 (2008). doi:

10.1016/S1748-0132(08)70014-8

Lynch, A. M., Guzzie, P. J., Bauer, D., Gocke, E., Itoh, S., Jacobs, A., Krul, C. A. M., Schepky,

A., Tanaka, N. & Kasper, P. Considerations on photochemical genotoxicity. II: Report of the

2009 International Workshop on Genotoxicity Testing Working Group. Mutat Res - Genet

Toxicol Environ Mutagen 723: 91–100 (2011). doi: 10.1016/j.mrgentox.2010.10.010

Magdolenova, Z., Collins, A., Kumar, A., Dhawan, A., Stone, V. & Dusinska, M. Mechanisms

of genotoxicity. A review of in vitro and in vivo studies with engineered nanoparticles.

Nanotoxicology 8: 233–278 (2014). doi: 10.3109/17435390.2013.773464.

Magkoufopoulou C., Claessen SM., Tsamou M., Jennen D.G., Kleinjans J.C., van Delft J.H.

A transcriptomics-based in vitro assay for predicting chemical genotoxicity in vivo.

Carcinogenesis 33(7):1421-9 (2012). doi: 10.1093/carcin/bgs182

Mahony, C., Bowtell, P., Huber, M., Kosemund, K., Pfuhler, S., Zhu, T., Barlow, S. & McMillan,

D. A. Threshold of toxicological concern (TTC) for botanicals - Concentration data analysis of

potentially genotoxic constituents to substantiate and extend the TTC approach to botanicals.

Food Chem Toxicol 138: 111182 (2020). doi: 10.1016/j.fct.2020.111182.

Makri A., Goveia M., Balbus J., Parkin R. Children’s susceptibility to chemicals: a review by

developmental stage. Journal of toxicology and environmental health. Part B 7(6):417-435

(2004). doi: 10.1080/10937400490512465.

Manová E., von Goetz N., Haurib U., Bogdala C., Hungerbühler K. Organic UV-filters in

personal care products in Switzerland: A survey of occurrence and concentrations. Int J Hyg

Environ Health 216(4):508–514 (2013). doi: 10.1016/j.ijheh.2012.08.003

Manová E., von Goetz N., Hungerbuehler K., Aggregate consumer exposure to UV filter

ethylhexyl methoxycinnamate via personal care products, Environ Int 74:249–257 (2015).

doi: doi: 10.1016/j.envint.2014.09.008

Martus, H. J., Froetschl, R., Gollapudi, B., Honma, M., Marchetti, F., Pfuhler, S., Schoeny,

R., Uno, Y., Yauk, C. & Kirkland, D. J. Summary of major conclusions from the 7th

_____________________________________________________________________________________________

125

International Workshop on Genotoxicity Testing (IWGT), Tokyo, Japan Mutat Res Gen Tox En

852: 5031348 (2020). doi: 10.1016/j.mrgentox.2020.503134.

Marx-Stoelting P., Adriaens E., Ahr HJ., Bremer S., Garthoff B., Gelbke H.P., Piersma A.,

Pellizzer C., Reuter U., Rogiers V., Schenk B., Schwengberg S., Seiler A., Spielmann H.,

Steemans M., Stedman D.B., Vanparys P., Vericat J.A., Verwei M., van der Water F., Weimer

M., Schwarz M. A review of the implementation of the embryonic stem cell test (EST). The

report and recommendations of an ECVAM/ReProTect Workshop. ATLA 37(3):313-28 (2009).

doi: 10.1177/026119290903700314.

Mascolo MG, Perdichizzi S, Vaccari M, Rotondo F, Zanzi C, Grilli S, Paparella M, Jacobs MN,

Colacci A. The Transformics Assay: First Steps for the Development of an Integrated Approach

to Investigate the Malignant Cell Transformation in vitro. Carcinogenesis 39(7):955-967

(2018). doi: 10.1093/carcin/bgy037

McArdle, M. E., Freeman, E. L., Staveley, J. P., Ortego, L. S., Coady, K. K., Weltje, L.,

Weyers, A., Wheeler, J. R. & Bone, A. J. Critical Review of Read‐Across Potential in Testing

for Endocrine‐Related Effects in Vertebrate Ecological Receptors. Environ Toxicol Chem 39(4):

739–753 (2020). doi: 10.1002/etc.4682

McNamara C., Rohan D., Golden D., Gibney M., Hall B., Tozer S., Safford B., Coroama M.,

Leneveu-Duchemin MC., Steiling W. Probabilistic modelling of European consumer exposure

to cosmetic products. Food Chem Toxicol 45(11):2086-96 (2007). doi:

10.1016/j.fct.2007.06.037.

Meek ME, Boobis AR, Crofton KM, Heinemeyer G, Raaij MV, Vickers C, Risk assessment of

combined exposure to multiple chemicals. A WHO/IPCS framework. Regul Toxicol Pharmacol

60:S1-S14 (2011). doi: 10.1016/j.yrtph.2011.03.010

Møller, P., Azqueta, A., Boutet-Robinet, E., Koppen, G., Bonassi, S., Milić, M., Gajski, G.,

Costa, S., Teixeira, J. P., Costa Pereira, C., Dusinska, M., Godschalk, R., Brunborg, G.,

Gutzkow, K. B., Giovannelli, L., Cooke, M. S., Richling, E., Laffon, B., Valdiglesias, V.,

Basaran, N., Del Bo’, C., Zegura, B., Novak, M., Stopper, H., Vodicka, P., Vodenkova, S., de

Andrade, V. M., Sramkova, M., Gabelova, A., Collins, A. & Langie, S. A. S. Minimum

Information for Reporting on the Comet Assay (MIRCA): recommendations for describing

comet assay procedures and results. Nature Protocols (2020). doi:10.1038/s41596-020-

0398-1.

Monopoli M.P., Aberg C., Salvati A., Dawson K.A. Biomolecular coronas provide the

biological identity of 9030 nanosized materials. Nat Nanotechnol 7:779-786 (2012). doi:

10.1038/nnano.2012.207.

Moore T.L., Rodriguez-Lorenzo L., Hirsch V., Balog S., Urban D., Jud C., Rothen-Rutishauser

B., Lattuada M., Petri-Fink A. Nanoparticle colloidal stability in cell culture media and impact

on cellular interactions. Chem Soc Rev 44(17):6287-305 (2015). doi: 10.1039/C4CS00487F

Moxon, T. E., Li, H., Lee, M. Y., Piechota, P., Nicol, B., Pickles, J., Pendlington, R., Sorrell, I.

& Baltazar, M. T. Application of physiologically based kinetic (PBK) modelling in the next

generation risk assessment of dermally applied consumer products. Toxicol Vitr 63: 1-12

(2020). doi:10.1016/j.tiv.2019.104746.

Müller L., Gocke E. The rise and fall of photomutagenesis. Mutat Res 752(2):67-71 (2013).

doi: 10.1016/j.mrrev.2013.02.002.

Munro IC., Ford RA., Kennepohl E., Sprenger JG. Correlation of structural class with no-

observed-effect levels: A proposal for establishing a threshold of concern. Food and Chem

Toxicol 34(9):829-867 (1996). doi: 10.1016/s0278-6915(96)00049-x.

 
 
 
 
 
_____________________________________________________________________________________________ 
126 
 
 
Nielsen E, Thorup I., Schnipper A., Hass U., Meyer O., Ladefoged O., Larsen J.C, Østergaard 
G and Larsen P.B. (ed. Danish Environmental Protection Agency). Children and the unborn 
child.  Exposure  and  susceptibility  to  chemical  substances  –  an  evaluation.  Environmental 
Project No. 589 (2001). 
 
Nijkamp  MM.,  Bokkers  BG.,  Bakker  MI.,  Ezendam  J.,  Delmaar  JE.,  Quantitative  risk 
assessment  of  the  aggregate  dermal  exposure  to  the  sensitizing  fragrance  geraniol  in 
personal care products and household cleaning agents. Regul Toxicol Pharmacol 73(1):9-18 
(2015). doi: 10.1016/j.yrtph.2015.06.004. 
 
Nishida  H., Hirota  M., Seto  Y., Suzuki  G., Kato  M., Kitagaki  M., Sugiyama  M., Kouzuki 
H., Onoue  S.  Non-animal  photosafety  screening  for  complex  cosmetic  ingredients  with 
photochemical and photobiochemical assessment tools, Regul Toxicol Pharmacol 72(3):578-
85 (2015). doi: 10.1016/j.yrtph.2015.05.029 
OECD 150 - OECD (2018) Revised Guidance Document 150 on Standardised Test Guidelines 
for Evaluating Chemicals for Endocrine Disruption. OECD Series on Testing and Assessment 
No. 150, OECD Publishing, Paris, doi: 10.1787/9789264304741-en.  
OECD 151 - OECD (2013a) Guidance document (GD) supporting OECD Test Guideline 443 
(OECD, 2012) on the Extended One Generation Reproductive Toxicity Test. OECD Series on 
Testing and Assessment No. 151, OECD Publishing, Paris, July 9, 2013. 
 
OECD 156 - OECD (2011a) Guidance Notes on Dermal Absorption. OECD Series on Testing 
and Assessment No.156, OECD Publishing Paris, August 18, 2011. 
 
OECD 160 - OECD (2018), Guidance document on the collection of eye tissues for histological 
evaluation and collection of data. Second Edition, OECD Series on Testing and Assessment, 
No.160, OECD Publishing, Paris, July 6, 2018. 
 
OECD 168 - OECD (2012b), The adverse outcome pathway for skin sensitization initiated by 
covalent  binding  to  proteins  Part  I:  Scientific  Evidence.  OECD  Series  on  testing  and 
assessment, No. 168, OECD Publishing, Paris, May 4, 2012. doi: 10.1787/9789264221444-
en. 
 
OECD  184  -  OECD  (2017),  Revised  Guidance  Document  on  Developing  and  Assessing 
Adverse  Outcome  Pathways,  OECD  Series  on  Testing  and  Assessment,  No.  184,  OECD 
Publishing, Paris, July 27, 2017 
 
OECD 194 - OECD (2014a), Guidance on grouping of chemicals, second edition. OECD Series 
on Testing & Assessment, No. 194, OECD Publishing, Paris, April 14, 2014. 
 
OECD 203 - OECD (2014b), New Guidance Document on an Integrated Approach to Testing 
and Assessment for Skin Irritation and Corrosion. OECD Series on Testing and Assessment, 
No. 203, OECD Publishing, Paris, July 11, 2014. 
 
OECD 211 - OECD (2017), Guidance Document for Describing Non-Guideline In Vitro Test 
Methods, OECD Series on Testing and Assessment, No. 211, OECD Publishing, Paris, April 13, 
2017, doi: 10.1787/9789264274730-en. 
 
OECD 214 - OECD (2015), Guidance document on The in vitro Syrian Hamster Embryo (SHE) 
Cell  Transformation  Assay.  OECD  Series  on  Testing  and  Assessment,  No.  214,  OECD 
Publishing, Paris, May 22, 2015. 

 
 
 
 
 
_____________________________________________________________________________________________ 
127 
 
OECD  229  -  OECD  (2012), Test  No.  229:  Fish  Short  Term  Reproduction  Assay,  OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  2,  OECD  Publishing, 
Paris, https://doi.org/10.1787/9789264185265-en. 
OECD 231 – OECD (2009), Test No. 231: Amphibian Metamorphosis Assay. OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  2,  OECD  Publishing,  Paris,  (OECD,  2009). 
http://doi:10.1787/9789264076242-en. 
OECD  233  -  OECD  (2018),  Users’  Handbook  Supplement  to  the  Guidance  Document  for 
Developing and Assessing AOPs. OECD Series on Testing & Assessment No. 233, Series on 
Adverse  Outcome  Pathways  No.  1,  OECD  Publishing,  Paris,  February  14,  2018. 
https://doi.org/10.1787/5jlv1m9d1g32-en 
 
OECD 240 - OECD (2015), Test No. 240: Medaka Extended One Generation Reproduction 
Test (MEOGRT), OECD Guidelines for the Testing of Chemicals, Section 2, OECD Publishing, 
Paris, https://doi.org/10.1787/9789264242258-en. 
 
OECD 241 – OECD (2015), The Larval Amphibian Growth and Development Assay (LAGDA), 
OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  2,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264242340-en. 
 
OECD  248  –  OECD  (2019),  Test  No.  248:  Xenopus  Eleutheroembryonic  Thyroid  Assay 
(XETA), OECD  Guidelines for the Testing  of Chemicals, Section 2, OECD Publishing, Paris, 
https://doi.org/10.1787/a13f80ee-en.  
 
OECD 255 - OECD (2017a), Guidance Document on the Reporting of Defined Approaches to 
be Used  Within Integrated Approaches to  Testing and Assessment.  Series  on  Testing  and 
Assessment, No. 255. Paris, 2017. https://doi.org/10.1787/9789264274822-en. 
 
OECD 256 – OECD (2017), Guidance Document on the Reporting of Defined Approaches and 
Individual  Information  Sources  to  be  Used  within  Integrated  Approaches  to  Testing  and 
Assessment (IATA) for Skin Sensitisation, OECD Series on Testing and Assessment, No. 256, 
OECD Publishing, Paris, https://doi.org/10.1787/9789264279285-en. 
 
OECD 263 - OECD (2019), Second Edition - Guidance Document on Integrated Approaches 
to Testing and Assessment (IATA) for Serious Eye Damage and Eye Irritation, OECD Series 
on  Testing  and  Assessment,  No.  263,  OECD  Publishing,  Paris,  July  25,  2019, 
https://doi.org/10.1787/84b83321-en. 
 
OECD 267 - OECD (2017), OECD Validation report for the international validation study on 
the IL-8 Luc Assay, OECD Series on Testing and Assessment No. 267, OECD Publishing Paris 
2017.  
 
OECD 401 – OECD 1987, Test No. 401: Acute Oral Toxicity, OECD Guidelines for the Testing 
of Chemicals, Section 4, OECD Publishing, Paris, last updated February 24, 1997 and deleted 
December 17, 2002. https://doi.org/10.1787/9789264040113-en. 
 
OECD 402 - OECD (2017), Test No. 402: Acute Dermal Toxicity, OECD Guidelines for the 
Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070585-en. 
 
OECD 403 - OECD (2009), Test No. 403: Acute Inhalation Toxicity, OECD Guidelines for the 
Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070608-en. 
 

 
 
 
 
 
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128 
 
OECD 404 - OECD (2015), Test No. 404: Acute Dermal Irritation/Corrosion, OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264242678-en 
 
OECD 405 - OECD (2020), Test No. 405: Acute Eye Irritation/Corrosion, OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264185333-en. 
 
OECD 406 - OECD (1992), Test No. 406: Skin Sensitisation, OECD Guidelines for the Testing 
of Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264070660-
en. 
 
OECD  407  -  OECD  (2008),  Test  No.  407:  Repeated  Dose  28-day  Oral  Toxicity  Study  in 
Rodents, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264070684-en 
 
OECD  408  -  OECD  (2018),  Test  No.  408:  Repeated  Dose  90-Day  Oral  Toxicity  Study  in 
Rodents, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264070707-en 
 
OECD 409 - OECD (1998), Test No. 409: Repeated Dose 90-Day Oral Toxicity Study in Non-
Rodents, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264070721-en 
 
OECD 410 - OECD (1981), Test No. 410: Repeated Dose Dermal Toxicity: 21/28-day Study, 
OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070745-en. 
 
OECD 411 - OECD (1981), Test No. 411: Subchronic Dermal Toxicity: 90-day Study, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070769-en 
 
OECD 412 - OECD (2018), Test No. 412: Subacute Inhalation Toxicity: 28-Day Study, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070783-en. 
 
OECD 413 - OECD (2018), Test No. 413: Subchronic Inhalation Toxicity: 90-day Study, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070806-en 
 
OECD  414  -  OECD  (2018),  Test  No.  414:  Prenatal  Developmental  Toxicity  Study,  OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070820-en. 
 
OECD  416  -  OECD  (2001),  Test  No.  416:  Two-Generation  Reproduction  Toxicity,  OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070868-en. 
 
OECD 417 - OECD (2010), Test No. 417: Toxicokinetics, OECD Guidelines for the Testing of 
Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264070882-en. 
 
OECD 420 - OECD (2002), Test No. 420: Acute Oral Toxicity - Fixed Dose Procedure, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264070943-en 
 
OECD 421 - OECD (2016), Test No. 421: Reproduction/Developmental Toxicity Screening 
Test,  OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264264380-en 

 
 
 
 
 
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129 
 
 
OECD 422 - OECD (2016), Test No. 422: Combined Repeated Dose Toxicity Study with the 
Reproduction/Developmental  Toxicity  Screening  Test,  OECD  Guidelines  for  the  Testing  of 
Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264264403-en. 
 
OECD 423 - OECD (2002), Test No. 423: Acute Oral toxicity  - Acute Toxic Class Method, 
OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071001-en. 
 
OECD 425 - OECD (2008), Test No. 425: Acute Oral Toxicity: Up-and-Down Procedure, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071049-en. 
 
OECD 427 - OECD (2004), Test No. 427: Skin Absorption: In Vivo Method, OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071063-en. 
 
OECD 428 - OECD (2004), Test No. 428: Skin Absorption: In Vitro Method, OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071087-en. 
 
OECD 429 - OECD (2010), Test No. 429: Skin Sensitisation: Local Lymph Node Assay, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071100-en. 
 
OECD 430 - OECD (2015), Test No. 430: In Vitro Skin Corrosion: Transcutaneous Electrical 
Resistance Test Method (TER), OECD Guidelines for the Testing of Chemicals, Section 4, OECD 
Publishing, Paris, https://doi.org/10.1787/9789264242739-en. 
 
OECD  431  -  OECD  (2019),  Test  No.  431:  In  vitro  skin  corrosion:  reconstructed  human 
epidermis (RHE) test method, OECD Guidelines for the Testing of Chemicals, Section 4, OECD 
Publishing, Paris, https://doi.org/10.1787/9789264264618-en 
 
OECD  432  -  OECD  (2019),  Test  No.  432:  In  Vitro  3T3  NRU  Phototoxicity  Test,  OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071162-en. 
 
OECD  433  - OECD  (2018),  Test  No. 433:  Acute  Inhalation  Toxicity:  Fixed Concentration 
Procedure, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264284166-en 
 
OECD 435 - OECD (2015), Test No. 435: In Vitro Membrane Barrier Test Method for Skin 
Corrosion, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264242791-en. 
 
OECD  436  -  OECD  (2009),  Test  No.  436:  Acute  Inhalation  Toxicity  –  Acute  Toxic  Class 
Method, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264076037-en. 
 
OECD  437  -  OECD  (2020),  Test  No.  437:  Bovine  Corneal  Opacity  and  Permeability  Test 
Method  for  Identifying  i)  Chemicals  Inducing  Serious  Eye  Damage  and  ii)  Chemicals  Not 
Requiring Classification for Eye Irritation or Serious Eye Damage, OECD Guidelines for the 
Testing  of  Chemicals,  Section  4,  Éditions  OCDE,  Paris,  26  June  2020, 
https://doi.org/10.1787/9789264203846-en. 
 
OECD 438 - OECD (2018), Test No. 438: Isolated Chicken Eye Test Method for Identifying i) 
Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for 

 
 
 
 
 
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130 
 
Eye Irritation or Serious Eye Damage, OECD Guidelines for the Testing of Chemicals, Section 
4, OECD Publishing, Paris, https://doi.org/10.1787/9789264203860-en. 
 
OECD  439  -  OECD  (2020),  Test  No.  439:  In  Vitro  Skin  Irritation:  Reconstructed  Human 
Epidermis Test Method, OECD Guidelines for the Testing of Chemicals, Section 4, Éditions 
OCDE, Paris, June 20, 2020, https://doi.org/10.1787/9789264242845-en. 
 
OECD 440 - OECD (2007), Test No. 440: Uterotrophic Bioassay in Rodents: A short-term 
screening  test  for  oestrogenic  properties,  OECD  Guidelines  for  the  Testing  of  Chemicals, 
Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264067417-en. 
 
OECD 442A - OECD (2010), Test No. 442A: Skin Sensitization: Local Lymph Node Assay: 
DA,  OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264090972-en 
 
OECD 442B - OECD (2018), Test No. 442B: Skin Sensitization: Local Lymph Node Assay: 
BrdU-ELISA  or  –FCM,  OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD 
Publishing, Paris, https://doi.org/10.1787/9789264090996-en. 
 
OECD  442C  -  OECD  (2020),  Test  No.  442C:  In  Chemico  Skin  Sensitisation : Assays 
addressing the Adverse Outcome Pathway key event on covalent binding to proteins, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  Éditions  OCDE,  Paris,  June  26,  2020, 
https://doi.org/10.1787/9789264229709-en. 
 
OECD 442D - OECD (2018), Test No. 442D: In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase 
Test Method, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264229822-en. 
 
OECD  442E  -  OECD  (2018),  Test  No.  442E:  In  Vitro  Skin  Sensitisation: In  Vitro  Skin 
Sensitisation assays addressing the Key Event on activation of dendritic cells on the Adverse 
Outcome  Pathway  for  Skin  Sensitisation,  OECD  Guidelines  for  the  Testing  of  Chemicals, 
Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264264359-en 
 
OECD 443 - OECD (2018), Test No. 443: Extended One-Generation Reproductive Toxicity 
Study,  OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264185371-en. 
 
OECD 451 - OECD (2018), Test No. 451: Carcinogenicity Studies, OECD Guidelines for the 
Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071186-en. 
 
OECD 452 - OECD (2018), Test No. 452: Chronic Toxicity Studies, OECD Guidelines for the 
Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071209-en 
 
OECD 453 - OECD (2018), Test No. 453: Combined Chronic Toxicity/Carcinogenicity Studies, 
OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264071223-en 
 
OECD  455  –  OECD  (2016),  Test  No.  455:  Performance-Based  Test  Guideline  for  Stably 
Transfected  Transactivation  In  Vitro  Assays  to  Detect  Estrogen  Receptor  Agonists  and 
Antagonists, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264265295-en 
 
OECD 456 - OECD (2011), Test No. 456: H295R Steroidogenesis Assay, OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264122642-en. 
 

 
 
 
 
 
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131 
 
OECD 460 – OECD (2017), Test No. 460: Fluorescein Leakage Test Method for Identifying 
Ocular Corrosives and Severe Irritants, OECD Guidelines for the Testing of Chemicals, Section 
4, OECD Publishing, Paris, https://doi.org/10.1787/9789264185401-en. 
 
OECD 471 - OECD (2020), Test No. 471: Bacterial Reverse Mutation Test, OECD Guidelines 
for  the  Testing  of  Chemicals,  Section  4,  Éditions  OCDE,  Paris,  June  26,  2020, 
https://doi.org/10.1787/9789264071247-en 
 
OECD 487 - OECD (2016), Test No. 487: In Vitro Mammalian Cell Micronucleus Test, OECD 
Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing,  Paris, 
https://doi.org/10.1787/9789264264861-en. 
OECD 488 – OECD (2020), Test No. 488: Transgenic Rodent Somatic and Germ Cell Gene 
Mutation Assays, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, 
Paris, https://doi.org/10.1787/9789264203907-en. 
OECD  491 -  OECD  (2020),  Test  No.  491: Short  Time  Exposure  In Vitro  Test Method  for 
Identifying  i)  Chemicals  Inducing  Serious  Eye  Damage  and  ii)  Chemicals  Not  Requiring 
Classification for Eye Irritation or Serious Eye Damage, OECD Guidelines for the Testing of 
Chemicals,  Section  4,  Éditions  OCDE,  Paris,  June  26,  2020,  
https://doi.org/10.1787/9789264242432-en. 
 
OECD  492  -  OECD  (2019),  Test  No.  492:  Reconstructed  human  Cornea-like  Epithelium 
(RhCE) test method for identifying chemicals not requiring classification and labelling for eye 
irritation or serious eye damage, OECD Guidelines for the Testing of Chemicals, Section 4, 
OECD Publishing, Paris, https://doi.org/10.1787/9789264242548-en. 
 
OECD  493  -  OECD  (2015),  Test  No.  493:  Performance-Based  Test  Guideline  for  Human 
Recombinant Estrogen Receptor (hrER) In Vitro Assays to Detect Chemicals with ER Binding 
Affinity, OECD  Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, 
https://doi.org/10.1787/9789264242623-en. 
 
OECD 494 - OECD (2019), Test No. 494: Vitrigel-Eye Irritancy Test Method for Identifying 
Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage, 
OECD  Guidelines  for  the  Testing  of  Chemicals,  Section  4,  OECD  Publishing, 
Paris, https://doi.org/10.1787/9f20068a-en. 
 
OECD 496 - OECD (2019), Test No. 496: In vitro Macromolecular Test Method for Identifying 
Chemicals Inducing Serious Eye Damage and Chemicals Not Requiring Classification for Eye 
Irritation or Serious Eye Damage, OECD Guidelines for the Testing of Chemicals, Section 4, 
OECD Publishing, Paris,  
https://doi.org/10.1787/970e5cd9-en. 
 
OECD 2004 - Guidance Document for the Conduct of Skin Absorption Studies. Document 
number ENV/JM/MONO (2004)2, Series on Testing and Assessment, No. 28, OECD Publishing, 
Paris, 5 March 2004, doi:  /10.1787/9789264078796-en 
 
OECD 2017 - OECD (2017), Guidance Document on the In vitro Bhas 42 Cell Transformation 
Assay (BHAS 42 CTA), OECD Series on Testing & Assessment,  No. 231, OECD Publishing, 
ENV/JM/MONO (2016)1, Paris, July 20, 2017. 
 
OECD 2017a - Revised Guidance Document on Developing and Assessing Adverse Outcome 
Pathways  Series  on  Testing  &  Assessment  No.  184.  2nd  edition.  ENV/JM/MONO  (2013)6. 
Paris, 2017. 
 
OECD  2017b  -  Guidance  Document  on  the  Reporting  of  Defined  Approaches  to be  Used 
Within Integrated Approaches to Testing and Assessment. Series on Testing and Assessment, 
No. 255. Paris, 2017. 

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132

OECD 2017c - Guidance Document on Considerations for Waiving or Bridging of mammalian

Acute Toxicity Tests. Series on Testing and Assessment No. 237, Paris, April 13,

2017.https://doi:10.1787/9789264274754-en.

OECD 2018 - Guidance Document on Inhalation Toxicity Studies, Second Edition, OECD

Series on Testing and Assessment, No. 39, OECD Publishing, Paris, July 6, 2018.

OECD 2018 - Users’ handbook supplement to the guidance document for developing and

assessing AOPs Series on Testing & Assessment No. 233 Series on Adverse Outcome

Pathways No. 1. ENV/JM/MONO (2016)12. Paris, 2018.

Oesch F., Fabian E., Oesch-Bartlomowicz B., Werner C., Landsiedel R. Drug-metabolizing

enzymes in the skin of man, rat, and pig. Drug Metab Rev 39(4):659-98, (2007).

Oesch F. and Hengstler J.G. Importance of Metabolism. Mechanistic Considerations Relevant

for Toxicological Regulation. In: Reichl FX. Schwenk M. (eds) Regulatory Toxicology. Springer,

Berlin, Heidelberg (2014a). doi : 10.1007/978-3-642-35374-1_71

Oesch F., Fabian E., Guth K., Landsiedel R. Xenobiotic-metabolizing enzymes in the skin of

rat, mouse, pig, guinea pig, man, and in human skin models. Arch Toxicol 88(12):2135-2190

(2014b). doi: 10.1007/s00204-014-1382-8

Oesch F., Fabian E., Landsiedel R. Xenobiotica-metabolizing enzymes in the lung of

experimental animals, man and in human lung models. Arch Toxicol 93(12):3419-3489

(2019). doi: 10.1007/s00204-019-02602-7

Onoue S, Seto Y, Sato H, Nishida H, Hirota M, Ashikaga T, Api AM, Basketter D, Tokura Y.

Chemical photoallergy: photobiochemical mechanisms, classification, and risk assessments.

J Dermatol Sci 85(1):4-11 (2017). doi: 10.1016/j.jdermsci.2016.08.005.

Parish, S. T., Aschner, M., Casey, W., Corvaro, M., Embry, M. R., Fitzpatrick, S., Kidd, D.,

Kleinstreuer, N. C., Lima, B. S., Settivari, R. S., Wolf, D. C., Yamazaki, D. & Boobis, A. An

evaluation framework for new approach methodologies (NAMs) for human health safety

assessment. Regul Toxicol Pharmacol 112:1-16 (2020). doi: 10.1016/j.yrtph.2020.104592.

Partosch F., Mielke H., Stahlmann R., Kleuser B., Barlow S., Gundert-Remy U. Internal

threshold of toxicological concern values: enabling route-to-route extrapolation. Arch Toxicol

89(6):941-948 (2014). doi: 10.1007/s00204-014-1287-6

Patel, A., Joshi, K., Rose, J., Laufersweiler, M., Felter, S. P. & Api, A. M. Bolstering the

existing database supporting the non-cancer Threshold of Toxicological Concern values with

toxicity data on fragrance-related materials. Regul Toxicol Pharmacol 116: 104718 (2020).

doi: 10.1016/j.yrtph.2020.104718

Patlewicz G., Simon TW., Rowlands JC., Budinsky RA., Becker RA. Proposing a scientific

confidence framework to help support the application of adverse outcome pathways for

regulatory purposes. Regul Toxicol Pharmacol 71:463-477 (2015). doi:

10.1016/j.yrtph.2015.02.011.

Patlewicz G., Casati S., Aschberger K., Bofill A., Basketter D., et al., Ability of NAM for skin

sensitization to detect pre- and pro-haptens. European Scientific and Technical Research

Report, JRC 100479 (2016). doi: 10.2788/01803

Patlewicz G., Helman G., Pradeep P, Shah I. Navigating through the minefield of read-across

tools. A review of in silico tools for grouping. Computational Toxicology 3: 1-18 (2017). doi:

10.1016/j.comtox.2017.05.003.

_____________________________________________________________________________________________

133

Pflaum M., Kielbassa C., Garmyn M., Epe B.G. Oxidative DNA damage induced by visible

light in mammalian cells; extent, inhibition by antioxidants and genotoxic effects. Mutat Res

408(2):137-144 (1998). doi: 10.1016/s0921-8777(98)00029-9.

Pfuhler, S., van Benthem, J., Curren, R., Doak, S. H., Dusinska, M., Hayashi, M., Heflich, R.

H., Kidd, D., Kirkland, D., Luan, Y., Ouedraogo, G., Reisinger, K., Sofuni, T., van Acker, F.,

Yang, Y. & Corvi, R. Use of in vitro 3D tissue models in genotoxicity testing: Strategic fit,

validation status and way forward. Report of the working group from the 7th International

Workshop on Genotoxicity Testing (IWGT). Mutat Res - Genet Toxicol Environ Mutagen 850–

851: 503135 (2020). doi: 10.1016/j.mrgentox.2020.503135

Pinalli, R., Croera, C., Theobald, A. & Feigenbaum, A. Threshold of toxicological concern

approach for the risk assessment of substances used for the manufacture of plastic food

contact materials. Trends Food Sci Technol 22: 523–534 (2011). doi:

10.1016/j.tifs.2011.07.001

Plošnik, A., Vračko, M. & Dolenc, M. S. Mutagenic and carcinogenic structural alerts and

their mechanisms of action. Archives of Industrial Hygiene and Toxicology 67: 169–182

(2016). doi: 10.1515/aiht-2016-67-2801.

Prieto P., Hoffmann S., Tirelli V., Tancredi F., González I., Bermejo M., De Angelis I. (2010)

An Exploratory Study of Two Caco-2 Cell Models for OralAbsorption: A Report on Their Within-

laboratory and Between-aboratory Variability, and Their Predictive Capacity. ATLA 38:367–

386 (2010). doi: 10.1177/026119291003800510.

Proksch, E. pH in nature, humans and skin. J Dermatol 45: 1044–1052 (2018). doi:

10.1111/1346-8138.14489.

Quadros ME., Marr LC. Silver nanoparticles and total aerosols emitted by nanotechnology-

related consumer spray products. Environ Sci and Technol 45(24):10713-10719 (2011). doi:

10.1021/es202770m

Reilly, L., Serafimova, R., Partosch, F., Gundert-Remy, U., Cortiñas Abrahantes, J., Dorne,

J. L. M. C. & Kass, G. E. N. Testing the thresholds of toxicological concern values using a new

database for food-related substances. Toxicol Lett 314: 117–123 (2019). doi:

10.1016/j.toxlet.2019.07.019

Reisinger, K., Dony, E., Wolf, T. & Maul, K. Hen’s Egg Test for Micronucleus Induction (HET-

MN). Methods Mol Biol 2031: 195–208 (2019). doi: 10.1007/978-1-4939-9646-9_10.

Renwick AG. Dorne JL., Walton K. An analysis of the need for an additional uncertainty

factor for infants and children. Regul toxicol pharmacol 31(3):286-296 (2000). doi:

10.1006/rtph.2000.1394

Renwick AG. Toxicokinetics in infants and children in relation to the ADI and TDI. Food Addit

Contam Suppl 15:17-35 (1998). doi: 10.1080/02652039809374612.

Rieder BO. Consumer exposure to certain ingredients of cosmetic products: The case for tea

tree oil. Food Chem Toxicol 108(Pta):326-338 (2017). doi: 10.1016/j.fct.2017.08.012

RIVM 2006. National Institute for Public Health and the Environment. Cosmetics Fact

Sheet. To assess the risks for the consumer. Updated version for ConsExpo 4, RIVM Report

320104001.

Rocks S. Pollard S., Dorey R., Levy L., Harrison P., Handy R. Comparison of risk assessment

approaches for manufactured nanomaterials, Defra, London (2008).

_____________________________________________________________________________________________

134

Rogiers V. "Validated" and "valid" alternative methods available today for testing of cosmetic

products and their ingredients. In: Safety Assessment of Cosmetics in the EU. Training

Course Vrije Universiteit Brussel, 7-12 April, Part 2, p.1 (2003).

Rogiers V. and Beken S. (Editors and authors) Alternative Methods to Animal experiments.

Actual status, development and approach in Belgium. VUBPress, Brussels. ISBN 90-5487-

264-0 (2000).

Rogiers, V., Benfenati, E., Bernauer, U., Bodin, L., Carmichael, P., Chaudhry, Q., Coenraads,

P. J., Cronin, M. T. D., Dent, M., Dusinska, M., Ellison, C., Ezendam, J., Gaffet, E., Galli, C.

L., Goebel, C., Granum, B., Hollnagel, H. M., Kern, P. S., Kosemund-Meynen, K., Ouédraogo,

G., Panteri, E., Rousselle, C., Stepnik, M., Vanhaecke, T., von Goetz, N. & Worth, A. The way

forward for assessing the human health safety of cosmetics in the EU - Workshop proceedings.

Toxicology 436: 152421 (2020). doi: 10.1016/j.tox.2020.152421

Rothe H., Fautz R., Gerber E., Neumann L., Rettinger K., Schuh W., Gronewold C. Special

aspects of cosmetic spray safety evaluations: Principles on inhalation risk Assessment.

Toxicol Lett 205(2):97 – 104 (2011). doi: 10.1016/j.toxlet.2011.05.1038.

Ruiz P., Sack A., Wampole M., Bobst S., Vracko M. Integration of in silico methods and

computational systems biology to explore endocrine-disrupting chemical binding with nuclear

hormone receptors. Chemosphere 178:99-109 (2017). doi:

10.1016/j.chemosphere.2017.03.026.

Russell B., Russell WMS., Burch RL. The principles of Humane Experimental Technique.

Methuen and Co Ltd, London (reprinted by the Universities Federation for Animal Welfare

UFAW, 1992, Potters Bar, Herts), UK, (1959).

Saadatmand M., Stone KJ., Vega VN., Felter S., Ventura S., Kasting G., Jaworska J. Skin

hydration analysis by experiment and computer simulations and its implications for diapered

skin. Skin Res Technol 23(4):500-513 (2017). doi: 10.1111/srt.12362

Safford B, Api AM., Barratt C., Comiskey D., Daly EJ., Ellis G., McNamara C., O’Mahony C.,

Robison S., Smith B., Thomas R., Tozer S., Use of an aggregate exposure model to estimate

consumer exposure to fragrance ingredients in personal care and cosmetic products. Regul

Toxicol Pharmacol 72(3):673–682 (2015). doi: 10.1016/j.yrtph.2015.05.017

Sakuratani, Y., Horie, M. & Leinala, E. Integrated Approaches to Testing and Assessment:

OECD Activities on the Development and Use of Adverse Outcome Pathways and Case

Studies. Basic Clin Pharmacol Toxicol (2018) doi: 10.1111/bcpt.12955.

Sanner T., Dybing E., Willems MI. and Kroese ED. A simple method for quantitative risk

assessment of non-threshold carcinogens based on the dose descriptor T25. Pharmacol

Toxicol 88(6):331-341 (2001). doi: 10.1034/j.1600-0773.2001.880608.x

Sanner T. and Dybing E., Comparison of carcinogenic and in vivo genotoxic potency

estimates. BCPT96(2):131-139(2005), doi: 10.1111/j.1742-7843.2005.pto960110.x

SANTE/2018/10591 - Note for agreement by Member States’s Competent Authorities in the

SCoPAFF: Phytopharmaceutical legislation section Guidance on dermal absorption. (2018)

doi: 10.2903/j.efsa.2017.4873.

Santonen T. Human biomonitoring in risk assessment: analysis of the current practice and

1st examples on the use of HBM in risk assessments of HBM4EU priority chemicals Deliverable

Report D 5.1 (2018).

_____________________________________________________________________________________________

135

Sasaki K., Huk A., El Yamani N., Tanaka N., Dusinska M.. Bhas 42 CELL TRANSFORMATION

ASSAY FOR GENOTOXIC AND NON-GENOTOXIC CARCINOGENS In: Genotoxicity and Repair.

A Practical Approach, Eds. L Maria Sierra, Isabel Gavaivago, Series: Methods in Pharmacology

and Toxicology, Springer Protocols, Humana Press, Springer New York, Heidelberg Dordrecht

London, XII, 483 p. 70 (2014).

SCCNFP/0068/98: Guidelines on the use of human volunteers in compatibility testing of

finished cosmetic products, adopted by the SCCNFP during the plenary session of 23 June

1999.

SCCNFP/0098/99: Status report on the inventory of cosmetic ingredients, approved by the

plenary session of the SCCNFP on 17 February 1999.

SCCNFP/0120/99: Opinion concerning the predictive testing of potentially cutaneous

sensitising cosmetic ingredients or mixtures of ingredients, adopted by the SCCNFP during

the 11

plenary session of 17 February 2000.

SCCNFP/0167/99: Basic Criteria for the in vitro assessment of percutaneous absorption of

cosmetic ingredients, adopted by the SCCNFP during the 8

plenary meeting of 23 June 1999.

SCCNFP/0245/99: Opinion concerning Basic Criteria of the protocols for the skin

compatibility testing of potentially cutaneous irritant cosmetic ingredients or mixtures of

ingredients on human volunteers, adopted by the SCCNFP during the plenary session of 8

December 1999.

SCCNFP/0299/00: Opinion on the 1

update of the inventory of ingredients employed in

cosmetic products (Section I), adopted by the SCCNFP during the 13

plenary session of 28

June 2000.

SCCNFP/0321/00: Notes of Guidance for Testing of Cosmetic Ingredients for Their Safety

Evaluation, 4

revision, adopted by the SCCNFP during the plenary meeting of 24 October

2000.

SCCNFP/0389/00: Opinion concerning the 1

update of the inventory of ingredients

employed in cosmetic products. Section II: perfume and aromatic raw materials, adopted by

the SCCNFP during the plenary session of 24 October 2000.

SCCNFP/0483/01: Opinion on the evaluation of potentially estrogenic effects of UV-filters,

adopted by the SCCNFP during the 17

plenary meeting of 12 June 2001.

SCCNFP/0557/02: Position statement on the calculation of the Margin of Safety of

ingredients incorporated in cosmetics which may be applied to the skin of children, adopted

by the SCCNFP during the 19

plenary meeting of 27 February 2002.

SCCNFP/0633/02: Updated Basic Requirements for toxicological dossiers to be evaluated

by the SCCNFP, adopted by the SCCNFP during the 22

plenary meeting of 17 December

2002.

SCCNFP/0669/03: Opinion concerning Dihydroxyindoline HBr (Colipa noA147), adopted by

the SCCNFP during the 23

plenary meeting of 18 March 2003.

SCCNFP/0690/03: Notes of Guidance for the testing of cosmetic ingredients and their

safety evaluation, adopted by the SCCNFP during the 25

plenary meeting of 20 October

2003.

SCCNFP/0807/04: Opinion concerning hair dyes without file submitted, adopted by the

SCCNFP on 23 April 2004 by means of the written procedure.

_____________________________________________________________________________________________

136

SCCP (Scientific Committee on Consumer Products), Memorandum on hair dye substances

and their skin sensitising properties, 19 December 2006

SCCP/0919/05 (Scientific committee on consumer products), Memorandum on the

classification and categorization of skin sensitizers and grading of test reactions, adopted by

the SCCP during the 5

plenary meeting of 20 September 2005, SCCP/0919/05

SCCP/0933/05 (Scientific committee on consumer products), Opinion on risk of ingredients

deriving from category 1-material and category 2-material as defined in Regulation

1774/2002 in cosmetic products, adopted by the SCCP during the 5

plenary meeting of 20

September 2005, SCCP/0933/05

SCCP/0949/05 (Scientific committee on consumer products), Opinion on biological effects

of ultraviolet radiation relevant to health with particular reference to sunbeds for cosmetic

purposes, adopted by the SCCP during the 8

plenary meeting of 20 June 2006,

SCCP/0949/05

SCCP/0959/05 (Scientific committee on consumer products), Review of the SCCNFP opinion

on Hair Dye Strategy in the light of additional information, adopted by the SCCP during the

plenary meeting of 20 June 2006, SCCP/0959/05

SCCP/0970/06 (Scientific committee on consumer products), Opinion on Basic Criteria for

the in vitro assessment of dermal absorption of cosmetic ingredients - updated February

2006, adopted by the SCCP during the 7

plenary meeting of 28 March 2006, SCCP/0970/06

SCCP/1017/06 (Scientific committee on consumer products), Opinion on parabens (Colipa

n° P82), adopted by the SCCP during the 9

plenary meeting of 10 October 2006,

SCCP/1017/06

SCCP/1086/07 (Scientific committee on consumer products), Opinion on homosalate

(Colipa n° S12), adopted by the SCCP during the 11

plenary meeting of 21 March 2007,

SCCP/1086/07

SCCP/1153/08 (Scientific committee on consumer products), Opinion on dermal

sensitisation quantitative risk assessment (citral, farnesol and phenylacetaldehyde), adopted

by the SCCP during the 16

plenary meeting of 24 June 2008, SCCP/1153/08

SCCP/1171/08 (Scientific committee on consumer products), SCCS, SCHER, SCENIHR Joint

Opinion on the Use of the Threshold of Toxicological Concern (TTC). Approach for Human

Safety Assessment of Chemical Substances with focus on Cosmetics and Consumer Products,

adopted 8 June 2012, SCCP/1171/08

SCCP/1183/08 (Scientific committee on consumer products), Opinion on parabens (Colipa

no P82), adopted by the SCCP during the 16th plenary of 24 June 2008, SCCP/1183/08

SCCP/1192/08 (Scientific committee on consumer products), Opinion on Triclosan (Colipa

no P32), adopted by the SCCP during the 19

plenary meeting of 21 January 2009,

SCCP/1192/08

SCCS/1270/09 (Scientific Committee on Consumer Safety), Opinion on resorcinol, 23

March 2010, SCCS/1270/09

SCCS/1241/10 (Scientific Committee on Consumer Safety), Opinion on cyclomethicone

D4/D5, 22 June 2010, SCCS/1241/10

SCCS/1291/10 (Scientific Committee on Consumer Safety), Opinion on o-aminophenol, 22

June 2010, SCCS/1291/10

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137

SCCS/1311/10 (Scientific Committee on Consumer Safety), Opinion on reaction products

of oxidative hair dye ingredients formed during hair dyeing process, 21 September 2010,

SCCS/1311/10

SCCS/1315/10 (Scientific Committee on Consumer Safety), Opinion on Melatonin, 23 March

2010, SCCS/1315/10

SCCS/1348/10 (Scientific Committee on Consumer Safety), Opinion on parabens, 14

December 2010, revision of 22 March 2011, SCCS/1348/10

SCCS/1358/10 (Scientific Committee on Consumer Safety), basic criteria for the in vitro

assessment of dermal absorption of cosmetic ingredients, 22 June 2010, SCCS/1358/10

SCCS/1392/10 (Scientific Committee on Consumer Safety), memorandum (addendum) on

the in vitro test EPISKIN™ for skin irritation testing, 14 December 2010, SCCS/1392/10

SCCS/1400/11 (Scientific Committee on Consumer Safety), Opinion on 6- amino-m-cresol,

2-Amino-5-methylphenol, 26-27 June 2012, SCCS/1400/11

SCCS/1414/11 (Scientific Committee on Consumer Safety), Opinion on triclosan,

ADDENDUM to the SCCP Opinion on Triclosan (SCCP/1192/08) from January 2009, 22 March

2011, SCCS/1414/11

SCCS/1443/11 (Scientific Committee on Consumer Safety), Opinion on p-

phenylenediamine, 26-27 June 2012, SCCS/1443/11

SCCS/1446/11 (Scientific Committee on Consumer Safety), Clarification on Opinion

SCCS/1348/10 in the light of the Danish clause of safeguard banning the use of parabens in

cosmetic products intended for children under three years of age, 10 October 2011,

SCCS/1446/11

SCCS/1459/11 (Scientific Committee on Consumer Safety), opinion on fragrance allergens

in cosmetic products, 13-14 December 2011, SCCS/1459/11

SCCS/1479/12 (Scientific Committee on Consumer Safety), Opinion on toluene-2,5-

diamine and its sulfate, 26-27 June 2012, SCCS/1479/12

SCCS/1481/12 (Scientific Committee on Consumer Safety), Opinion on kojic acid, 26-27

June 2012, SCCS/1481/12

SCCS/1484/12 (Scientific Committee on Consumer Safety), Guidance on safety assessment

of nanomaterials in cosmetics, adopted by the SCCS at its 15th plenary meeting of 26-27

June 2012, SCCS/1484/12

SCCS/1486/12 (Scientific Committee on Consumer Safety), Opinion on NDELA in cosmetic

products and nitrosamines in balloons, adopted at its 15

plenary meeting on 26-27 June

2012, SCCS/1486/12

SCCS/1501/12 (Scientific Committee on Consumer Safety), The SCCS's Notes of Guidance

for the testing of cosmetic ingredients and their safety evaluation - 8

revision, adopted by

the SCCS during the 17

plenary meeting of 11 December 2012, SCCS/1501/12

_____________________________________________________________________________________________

138

SCCS/1509/13 (Scientific Committee on Consumer Safety), Memorandum on hair dye

Chemical Sensitisation, adopted by the SCCS at its 18th Plenary meeting of 26 February

2013, SCCS/1509/13

SCCS/1512/13 (Scientific Committee on Consumer Safety), Opinion on Zinc pyrithione, 18

June 2013, revision of 18 June 2014, SCCS/1512/13

SCCS/1513/13 (Scientific Committee on Consumer Safety), Opinion on 3-Benzylidene

camphor (Colipa n° S61), adopted by the SCCS during the 2nd plenary meeting of 18 June

2010, SCCS/1513/13

SCCS/1514/13 (Scientific Committee on Consumer Safety), Opinion on Parabens - Updated

request for a scientific opinion on propyl-and butylparaben (Colipa no P82), adopted by

written procedure on 3 May 2013, SCCS/1514/13

SCCS/1524/13 (Scientific Committee on Consumer Safety), Memorandum on "Relevance

and Quality of Data in Safety Dossiers on Nanomaterials", 12 December 2013, revision of 27

March 2014, SCCS/1524/13

SCCS/1532/14 (Scientific Committee on Consumer Safety), Addendum to the SCCS's Notes

of Guidance (NoG) for the Testing of Cosmetic Ingredients and their Safety Evaluation, 8th

Revision, adopted by the SCCS by written procedure on 9 April 2014, revision of 22 October

2014, SCCS/1532/14

SCCS/1533/14 (Scientific Committee on Consumer Safety), Opinion on 2-(4-(2-(4-

Diethylamino-2-hydroxy-benzoyl)-benzoyl)-piperazine-1-carbonyl)-phenyl)- (4-

diethylamino-2-hydroxyphenyl)-methanone (HAA299) as UV filter in sunscreen products.

The SCCS adopted this opinion at its 6th plenary meeting, revision of 23 September 2014,

SCCS/1533/14

SCCS/1538/14 (Scientific Committee on Consumer Safety), Opinion on the safety of the

use of formaldehyde in nail hardeners, written procedure 7 November 2014, revision of 16

December 2014, SCCS/1538/14

SCCS/1539/14 (Scientific Committee on Consumer Safety), Opinion for clarification of the

meaning of the term "sprayable applications/products" for the nano forms of Carbon Black CI

77266, Titanium Oxide and Zinc Oxide, adopted by the SCCS at its 7th plenary meeting on

23 September 2014, SCCS/1539/14

SCCS/1544/14 (Scientific Committee on Consumer Safety), Memorandum on Endocrine

Disruptors, 16 December 2014, SCCS/1544/14

SCCS/1546/15 (Scientific Committee on Consumer Safety), Opinion on 2,2’-methylene-

bis-(6(2H-benzotriazol-2-yl)-4-(1,1,3,3-tetramethylbutyl)phenol), 25 March 2015,

SCCS/1546/15

SCCS/1567/15 (Scientific Committee on Consumer Safety), Memorandum on use of Human

Data in risk assessment of skin sensitization, 15 December 2015, SCCS/1567/15

SCCS/1578/16 (Scientific Committee on Consumer Safety), Memorandum on the use of In

silico Methods for Assessment of Chemical Hazard, 6 October 2016, SCCS/1578/16

SCCS/1589/17 (Scientific Committee on Consumer Safety) Opinion on Skin Sensitisation

Quantitative Risk Assessment for Fragrance Ingredients (QRA2), 30 July 2018,

SCCS/1589/17

_____________________________________________________________________________________________

139

SCCS/1594/18 (Scientific Committee on Consumer Safety), Opinion on the safety of

cosmetic ingredients Phenylene Bis-Diphenyltriazine (CAS No 55514-22-2) - S86 -Submission

II, preliminary version of 21-22 February 2018, final version of 30 July 2018, SCCS/1594/18

SCCS/1595/18 (Scientific Committee on Consumer Safety), Final Opinion on

Styrene/Acrylates copolymer (nano) and Sodium styrene/Acrylates copolymer (nano), 22

February 2019, SCCS/1595/18

SCCS/1596/2018 (Scientific Committee on Consumer Safety), Opinion on Colloidal Silver,

24-25 October 2018, SCCS/1596/2018

SCCS/1602/18 (Scientific Committee on Consumer Safety), The SCCS's Notes of Guidance

for the testing of cosmetic ingredients and their safety evaluation - 10

revision, adopted by

the SCCS during the plenary meeting of 2’-25 October 2018, SCCS/1602/18

SCCS/1606/2019 (Scientific Committee on Consumer Safety), Opinion on solubility of

Synthetic Amorphous Silica (SAS), 20-21 June 2019, SCCS/1606/2019. Corrigendum of 6

December 2019.

SCCS/1611/19 (Scientific Committee on Consumer Safety), Guidance on the Safety

Assessment of Nanomaterials in Cosmetics, 30-31 October 2019, SCCS/1611/19

SCCS/1613/19 (Scientific Committee on Consumer Safety), Scientific advice on the safety

of aluminium in cosmetic products, plenary meeting 3-4 March 2020, submission II,

SCCS/1613/19

SCCS/1617/20 (Scientific Committee on Consumer Safety), Scientific advice on the safety

of titanium dioxide used in cosmetic products that lead to exposure by inhalation, written

procedure 6 October 2020, SCCS/1617/20

SCCS/1618/2020 (Scientific Committee on Consumer Safety), Scientific advice on the

safety of nanomaterials in cosmetics, 6 October 2020, SCCS/1618/2020

SCCS/1627/21 (Scientific Committee on Consumer Safety), Scientific advice on the safety

of octocrylene in cosmetic products, adopted by written procedure on 15 January 2021,

SCCS/1627/21

SCCS Expert Methodologies meeting in Brussels on May 25, 2011, with P Langguth

(University of Mainz, DE); L Turco (University of Barcelona, ES) and P Artursson (Uppsala

University, SE).

SCENIHR (Scientific Committee on Emerging and Newly Identified Health Risks), Risk

assessment of products of nanotechnologies, 19 January 2009.

Schenk B., Weimer M., Bremer S., van der Burg B., Cortvrindt R., Freyberger A., Lazzari G.,

Pellizzer C., Piersma A., Schäfer RW., Seiler A., Witters H., Schwarz M. The ReProTect

Feasibility Study, a novel comprehensive in vitro approach to detect reproductive toxicants.

Reprod Toxicol 30(1):200-218 (2010). doi: 10.1016/j.reprotox.2010.05.012

Schins, R. P. F. & Knaapen, A. M. Genotoxicity of poorly soluble particles. Inhal Toxicol

19(1): 189–198 (Taylor & Francis, 2007). doi: 10.1080/08958370701496202

Schmitz-Spanke, S. Toxicogenomics – What added Value Do These Approaches Provide for

Carcinogen Risk Assessment? Environ Res 173: 157–164 (2019).

Schneider, M., Pons, J.-L., Labesse, G. & Bourguet, W. In Silico Predictions of Endocrine

Disruptors Properties. Endocrinology 160(11): 2709–2716 (2019). doi: 10.1210/en.2019-

00382.

_____________________________________________________________________________________________

140

Schüürmann, G., Ebert, R. U., Tluczkiewicz, I., Escher, S. E. & Kühne, R. Inhalation

threshold of toxicological concern (TTC) - Structural alerts discriminate high from low

repeated-dose inhalation toxicity. Environ Int 88: 123–132 (2016). doi:

10.1016/j.envint.2015.12.005

Scott L., Eskes C., Hoffmann S., Adriaens E., Alepée N., Bufo M., Clothier R., Facchini D.,

Faller C., Guest R., Harbell J., Hartung T., Kamp H., Varlet B.L., Meloni M., McNamee P.,

Osborne R., Pape W., Pfannenbecker U., Prinsen M., Seaman C., Spielmann H., Stokes W.,

Trouba K., Berghe C.V., Van Goethem F., Vassallo M., Vinardell P., Zuang V. A proposed eye

irritation testing strategy to reduce and replace in vivo studies using Bottom-Up and Top-

Down approaches. Toxicol In vitro 24(1):1-9 (2010). doi: 10.1016/j.tiv.2009.05.019

Serafimova R., Gatnik M.F. and Worth A. Review of QSAR models and software tools for

predicting genotoxicity and carcinogenicity, EUR 24427 EN – 2010, European Commission

Joint Research Centre, Institute for Health and Consumer Protection, Italy, (2010). doi:

10.2788/26123

Serra, S., Vaccari, M., Mascolo, M. G., Rotondo, F., Zanzi, C., Polacchini, L., Wagner, C. B.,

Kunkelmann, T., Perschbacher, S., Poth, A., Grilli, S., Jacobs, M. N. & Colacci, A. Hazard

assessment of air pollutants: The transforming ability of complex pollutant mixtures in the

Bhas 42 cell model. ALTEX 36: 623–633 (2019). doi: 10.14573/altex.18121

Shah D, Paruchury S, Matta M, Chowan G, Subramanian M, Saxena A, Soars MG, Herbst J,

Haskell R, Marathe P, Mandlekar S, A systematic evaluation of solubility enhancing excipients

to enable the generation of permeability data for poorly soluble compounds in Caco-2 model,

Drug Metab Lett 2014 8(2):109-118 (2014). doi: 10.2174/1872312808666141127113055.

Šimon P., Joner E. Conceivable interactions of biopersistent nanoparticles with food matrix

and living systems following from their physicochemical properties. Journal Food Nutrit Res

47:51-59 (2008).

Simon, T. W., Zhu, Y., Dourson, M. L. & Beck, N. B. Bayesian methods for uncertainty factor

application for derivation of reference values. Regul Toxicol Pharmacol 80: 9–24 (2016). doi:

10.1016/j.yrtph.2016.05.018.

Smith MT, Guyton KZ, Gibbons CF, Fritz JM, Portier CJ, Rusyn I, DeMarini DM, Caldwell JC,

Kavlock RJ, Lambert PF, Hecht SS, Bucher JR, Stewart BW, Baan RA, Cogliano VJ, Straif K.

Key Characteristics of Carcinogens as a Basis for Organizing Data on Mechanisms of

Carcinogenesis. Environ Health Perspect 124(6):713-721 (2016). doi: 10.1289/ehp.1509912.

Smith, M. T. Key characteristics of carcinogens. Tumour site concordance and mechanisms

of carcinogenesis. IARC Scientific Publication, (165), 85-91. (2019).

Spielmann H., Balls M., Dupuis J., Pape W.J.W., De Silva O., Holzhütter H.G., Gerberick F.,

Liebsch M., Lowell W.W., and Pfannenbecker V. A Study on UV-filter chemicals from Annex

VII of the European Union Directive 76/768/EEC in the in vitro NRU Phototoxicity test. ATLA

26:679-708 (1998).

Steiblen, G., Benthem, J. van & Johnson, G. Strategies in genotoxicology: Acceptance of

innovative scientific methods in a regulatory context and from an industrial perspective. Mutat

Res - Genet Toxicol Environ Mutagen 853: 503171 (2020). doi:

10.1016/j.mrgentox.2020.503171

Steiling W, Bascompta M, Carthew P et al. Principle considerations for the risk assessment

of sprayed consumer products. Toxicol Lett 227:41–49 (2014). doi:

10.1016/j.toxlet.2014.03.005.

_____________________________________________________________________________________________

141

Steiling W., Almeida JF., Assaf, Vandecasteele H., Gulpen S., Kawamoto T., O’Keeffe L.,

Pappa G., Rettinger K., Rothe H., and Bowden A., Principles for the safety evaluation of

cosmetic powders. Toxicol Lett 297:8-18 (2018). doi: 10.1016/j.toxlet.2018.08.011

Steiling W., Buttgereit P., Hall B, O’Keeffe L, Safford B, Tozer S, Coroama M. Skin Exposure

to Deodorants / Antiperspirants in Aerosol Form. Food Chem Toxicol 50(6):2206-2215

(2012). doi: 10.1016/j.fct.2012.03.058.

Stenberg C., Larkö O. Sunscreen application and its importance for the sun protection factor.

Arch Dermatol 121(11):1400-1402 (1985).

Strittholt CA, McMillan DA, He T, Baker RA, Barker ML., A randomized clinical study to assess

ingestion of dentifrice by children. Regul Toxicol Pharmacol 75:66-71 (2016). doi:

10.1016/j.yrtph.2015.12.008.

Takenaka T., Kazuki K., Harada N., Kuze J.,Chiba M., Iwao., Matsunaga T., Abe S., Oshimura

M., Kazuki Y. Development of Caco-2 cells co-expressing CYP 3A4 and NADPH-cytochrome

P450 using a human artificial chromosome for the prediction of intestinal extraction ratio of

CYP 3A4 substrates, Drug Metab Pharmacokinet 32(1):61-68 (2017). doi:

10.1016/j.dmpk.2016.08.004

Thomas S., Brightman T.S. F., Gill H., Lee S., Pufong B. Simulation modelling of human

intestinal absorption using Caco-2 permeability and kinetic solubility data for early drug

discovery. J Pharm Sci 97(10):4557-4574 (2008). doi: 10.1002/jps.21305.

Tluczkiewicz, I., Buist, H. E., Martin, M. T., Mangelsdorf, I. & Escher, S. E. Improvement of

the Cramer classification for oral exposure using the database TTC RepDose - A strategy

description. Regul Toxicol Pharmacol 61(3): 340–350 (2011). doi:

10.1016/j.yrtph.2011.09.005.

Tollefsen KE., Scholz S., Cronin MT. et al. Applying Adverse Outcome Pathways (AOP) to

support Integrated Approaches to Testing and Assessment (IATA). Reg Tox Pharm 70(3):629-

640 (2014). doi: 10.1016/j.yrtph.2014.09.009.

Tozer SA., Kelly S., O'Mahony C., Daly EJ., Nash JF. Aggregate exposure modelling of zinc

pyrithione in rinse-off personal cleansing products using a person-orientated approach with

market share refinement. Food Chem Toxicol 83:103-110 (2015). doi:

10.1016/j.fct.2015.06.005.

Turco L., Catone T., Caloni F., Di Consiglio E., Testai E., Stammati A. Caco-2/TC7 cell line

characterization for intestinal absorption: How reliable is this in vitro model for the prediction

of the oral dose fraction absorbed in human? Toxicol in vitro 25(1):13-20 (2011). doi:

10.1016/j.tiv.2010.08.009.

UNEP/WHO: State of the Science of Endocrine disrupting Chemicals; Edited by Ake

Bergman, Jerrold J. Heindel, Susan Jobling, Karen A. Kidd and R. Thomas Zoeller. United

Nations Environment Programme and the World Health Organization (2013) ISBN: 978-92-

807-3274-0 available at: http://www.who.int/ceh/publications/endocrine/en (Consulted

December 2020). Federal Register 51: 33992-34003 (1986).

Urbisch D, Becker M, Honarvar N, Kolle SN, Mehling A, Teubner W, Wareing B, Landsiedel

R. Assessment of Pre- and Pro-haptens Using Nonanimal Test Methods for Skin Sensitization.

Chem Res Toxicol 16;29(5):901-913 (2016). doi: 10.1021/acs.chemrestox.6b00055.

Urbisch D., Mehling A., Guth K., Ramirez T., Honarvar N., Kolle S., Landsiedel R., Jaworska

J., Kern PS., Gerberick F., Natsch A., Emter R., Ashikaga T., Miyazawa M., Sakaguchi H.,

Assessing skin sensitization hazard in mice and men using non-animal test methods, Regul

Toxicol Pharmacol 71(2):337-351 (2015). doi: 10.1016/j.yrtph.2014.12.008.

_____________________________________________________________________________________________

142

US EPA Exposure Factors Handbook Doc. EPA/600/P-95/002Fa. Office of Research and

Development, U.S. Environmental Protection Agency, Washington, DC (1997).

US EPA Health Effect Test Guidelines. Dermal Penetration. US Environmental Protection

Agency (EPA). Doc. EPA 712-C-96-350, Washington, DC (1996).

U.S. EPA Exposure Factors Handbook 2011 Edition (Final Report), EPA/600/R-09/052F. U.S.

Environmental Protection Agency, Washington, DC, (2011).

https://cfpub.epa.gov/ncea/risk/recordisplay.cfm?deid=236252

USP 38 and USP 38 – NF 33: The Pharmacopeia of the United States of America (USP),

Thirty-Eighth Revision and the National Formulary (NF) Thirty-Third Edition – General Notices

and Requirements. Available at:

https://www.uspnf.com/sites/default/files/usp_pdf/EN/USPNF/usp-nf-

notices/usp38_nf33_gn.pdf

Vaccari M., Mascolo MG., Rotondo F., Morandi E., Querciolo D., Perdichizzi S., Zanzi C., Serra

S., Poluzzi V., Angelini P., Grilli S., Colacci A. Identification of pathway-based toxicity in the

BALB/c 3T3 cell model. Toxicol in vitro, 29(6):1240-1253 (2014). doi:

10.1016/j.tiv.2014.10.002

Van de Sandt JJM., van Burgsteden JA., Carmichael PL., Dick I., Kenyon S., Korinth G.,

Larese F., Limasset JC., Maas WJM., Montomoli L., Nielsen JB., Payan J-P., Robinson E.,

Sartorelli P., Schaller KH., Wilkinson SC., Williams FM. In vitro predictions of skin absorption

of caffeine, testosterone, and benzoic acid: a multi-centre comparison study. Regul Toxicol

Pharmacol 39(3):271-281 (2004). doi: 10.1016/j.yrtph.2004.02.004.

Van Der Ven, L. T. M., Rorije, E., Sprong, R. C., Zink, D., Derr, R., Hendriks, G., Loo, L. H.

& Luijten, M. A Case Study with Triazole Fungicides to Explore Practical Application of Next-

Generation Hazard Assessment Methods for Human Health. Chem Res Toxicol 33: 834-848

(2020). doi: 10.1021/acs.chemrestox.9b00484.

van Ravenzwaay, B., Jiang, X., Luechtefeld, T. & Hartung, T. The Threshold of Toxicological

Concern for prenatal developmental toxicity in rats and rabbits. Regul Toxicol Pharmacol 88:

157–172 (2017). doi: 10.1016/j.yrtph.2012.06.004.

Van Vliet E, Zang Q, Petersohn D. Non-animal methods to predict skin sensitization (II): an

assessment of defined approaches. Crit Rev Toxicol 48(5):359-374, (2018). doi:

10.1080/10408444.2018.1429386

Vedani A, Dobler M, Smieško M. VirtualToxLab - a platform for estimating the toxic potential

of drugs, chemicals and natural products. Toxicol Appl Pharmacol 1; 261(2):142-53 (2012).

doi: 10.1016/j.taap.2012.03.018.

Visscher M., Odio M., Taylor T., White T., Sargent S., Sluder L., Smith L., Flower T., Mason

B., Rider M., Huebner A., Bondurant P. Skin Care in the NICU Patient: Effects of Wipes versus

Cloth and Water on Stratum Corneum Integrity. Neonatology 96(4):226–234 (2009). doi:

10.1159/000215593.

Visscher, M. O. & Narendran, V. Imaging reveals distinct textures at three infant skin sites

and reflects skin barrier status. Ski Res Technol srt.12925 (2020a) doi: 10.1111/srt.12925.

Visscher, M. O., Adam, R., Brink, S. & Odio, M. Newborn infant skin: Physiology,

development, and care. Clin Dermatol 33(3):271-80 (2015). doi:

10.1016/j.clindermatol.2014.12.003

_____________________________________________________________________________________________

143

Visscher, M. O., Carr AN, Winget J, Huggins T, Bascom CC, Isfort R, Lammers K, Narendran

V. Correction: Biomarkers of neonatal skin barrier adaptation reveal substantial differences

compared to adult skin. Pediatr Res (2020b). doi: 10.1038/s41390-020-01147-1

Vuorinen A., Odermatt A., Schuster D. In silico methods in the discovery of endocrine

disrupting chemicals, J. Steroid Biochem Mol Biol 137:18–26 (2013). doi:

10.1016/j.jsbmb.2013.04.009.

Wareing, B., Kolle, S. N., Birk, B., Alépée, N., Haupt, T., Kathawala, R., Kern, P. S., Nardelli,

L., Raabe, H., Rucki, M., Ryan, C. A., Verkaart, S., Westerink, W. M. A., Landsiedel, R. &

Natsch, A. The kinetic direct peptide reactivity assay (kDPRA): Intra- and inter-laboratory

reproducibility in a seven-laboratory ring trial. ALTEX 37: 639–651 (2020). doi:

10.14573/altex.2004291

Wareing, B., Urbisch, D., Kolle, S. N., Honarvar, N., Sauer, U. G., Mehling, A. & Landsiedel,

R. Prediction of skin sensitization potency sub-categories using peptide reactivity data. Toxicol

Vitr 45: 134–145 (2017). doi: 10.1016/j.tiv.2017.08.015.

White, P. A., Luijten, M., Mishima, M., Cox, J. A., Hanna, J. N., Maertens, R. M. & Zwart, E.

P. In vitro mammalian cell mutation assays based on transgenic reporters: A report of the

International Workshop on Genotoxicity Testing (IWGT). Mutat Res - Genet Toxicol Environ

Mutagen 847: 403039 (2019). doi: 10.1016/j.mrgentox.2019.04.002

WHO (World Health Organisation) IPCS risk assessment terminology. Harmonization Project

Document No. 1. Geneva (2004).

WHO (World Health Organisation) Kielhorn J., Melching-Kollmu S., Mangelsdorf I. Dermal

Absorption. WHO / IPCS Environmental Health Criteria 235, (2006), accessible through

http://www.inchem.org/documents/ehc/ehc/ehc235.pdf (consulted December 2020).

WHO (World Health Organization) Assessing human health risks of chemicals: derivation of

guidance values for health-based exposure limits. Environmental Health Criteria, 170, WHO,

Geneva (1994).

WHO (World Health Organization) Evaluation of certain food additives and contaminants

(Forty-sixth report of the Joint FAO/WHO Expert Committee on Food Additives). WHO

Technical Report Series, No. 868 (1997).

WHO (World Health Organization) International Programme on Chemical Safety

Harmonization Project (IPCS). Characterization and application of physiologically based

pharmacokinetic models in risk assessment. Harmonization Project Document No. 9. (2010).

WHO (World Health Organization) Principles for the assessment of Risks to human health

from exposure to chemicals. Environmental Health Criteria, 210, WHO, Geneva (1999).

WHO/IPCS (World Health Organization) International Programme on Chemical Safety),

2002. Global Assessment of the State-of-the-Science of Endocrine Disruptors. Journal (2002).

Available online: www.who.int/ipcs/publications/new_issues/endocrine_disruptors/en/.

Consulted December 2020.

WHO/UNEP State of the science of endocrine disrupting chemicals, ISBN: 978-92-807-

3274-0 (UNEP) and 978 92 4 150503 1 (2012).

Wiegand C., Hewitt NJ., Merk HF., Reisinger K. Dermal xenobiotix metabolism: a comparison

between native human skin, four in vitro skin test systems and a liver system. Skin Pharmacol

Physiol 27(5):263-275 (2014). doi: 10.1159/000358272.

_____________________________________________________________________________________________

144

Wilde, E. C., Chapman, K. E., Stannard, L. M., Seager, A. L., Brüsehafer, K., Shah, U. K.,

Tonkin, J. A., Brown, M. R., Verma, J. R., Doherty, A. T., Johnson, G. E., Doak, S. H. &

Jenkins, G. J. S. A novel, integrated in vitro carcinogenicity test to identify genotoxic and

non-genotoxic carcinogens using human lymphoblastoid cells. Arch Toxicol. 92(2): 935-951

(2018) doi: 10.1007/s00204-017-2102-y.

Wilk-Zasadna I., Bernasconi C., Pelkonen O., Coecke S. Biotransformation in vitro: An

essential consideration in the quantitative in vitro-to-in vivo extrapolation (QIVIVE) of toxicity

data. Toxicology 332:8-19 (2014). doi: 10.1016/j.tox.2014.10.006

Williams, R. V., DeMarini, D. M., Stankowski, L. F., Escobar, P. A., Zeiger, E., Howe, J.,

Elespuru, R. & Cross, K. P. Are all bacterial strains required by OECD mutagenicity test

guideline TG471 needed? Mutat Res - Genet Toxicol Environ Mutagen 848: 503081 (2019).

doi: 10.1016/j.mrgentox.2019.503081

Worth A.P. and Balls M. The importance of the prediction model in the development and

validation of alternative tests. ATLA 29:135-143 (2001). doi:

10.1177/026119290102900210.

Yang C., Barlow SM., Muldoon, Jacobs KL., Vitcheva V., Boobis AR., Felter SP., Arvidson KB.,

Keller D., Cronin MTD., Enoch S., Worth A., Hollnagel HM. Thresholds of Toxicological Concern

for cosmetics-related substances: New database, thresholds, and enrichment of chemical

space. Food Chem Toxicol 109(1):170-193 (2017). doi: 10.1016/j.fct.2017.08.043

Yourick JJ., Koenig ML., Yourick DL., Bronaugh RL. Fate of chemicals in skin after dermal

application: does the in vitro skin reservoir affect the estimate of systemic absorption? Toxicol

Appl Pharmacology 195(3):309-320 (2004). doi: 10.1016/j.taap.2003.07.015.

Zielhuis R.L. Recent and potential advances applicable to the protection of workers` health-

Biological monitoring in Berlin, A. Yodaiken RE., Henman BA., (Eds.) Assessment of toxic

agents at the workplace - roles of ambient and biological monitoring. Martinus Nijhoff

Publishers, Boston, (1984).

Electronic resources:

http://ec.europa.eu/health/scientific_committees/docs/rules_procedure_2013_en.pdf

http://ec.europa.eu/health/scientific_committees/consumer_safety/requests/index_en.htm

http://ec.europa.eu/health/scientific_committees/consumer_safety/opinions/index_en.htm

http://ec.europa.eu/health/scientific_committees/consumer_safety/index_en.htm,

http://echa.europa.eu/documents/10162/13639/alternatives_test_animals_2014_en.pdf

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APPENDIX 1: INFORMATION ON REGULATION (EC) NO 1223/2009

AND THE SCCS

1. INTRODUCTION TO COSMETIC REGULATION (EC) No 1223/2009

Since July 2013, Regulation (EC) No 1223/2009 harmonises the safety of cosmetics within

the Member States, simplifies procedures and streamlines terminology. The most significant

changes introduced by the Cosmetic Regulation include:

(1) Strengthened safety requirements for cosmetic products Manufacturers need to

follow specific requirements in the preparation of a product safety report prior to

placing a product on the market.

(2) Introduction of the notion of a “responsible person” (RP)

Only cosmetic products for which a legal or natural person is designated within the EU

as a “responsible person” can be placed on the market. The Cosmetics Regulation

allows the precise identification of the RP and clearly outlines his/her obligations.

(3) Centralised notification of all cosmetic products placed on the EU market

The RP (mostly the manufacturer) will need to send the

Product notification only once via the EU Cosmetic Product Notification Portal (CPNP).

(4) Introduction of reporting serious undesirable effects

(SUE)

A RP and a distributor have the obligation to notify serious undesirable effects to

national authorities. The authorities will also collect information coming from end

users and health professionals. They will be obliged to share the information with

other EU countries. More information on reporting of SUE.

(5) New rules for the use of nanomaterials in cosmetic products

(6) A set of requirements for CMR (carcinogenic, mutagenic, toxic for reproduction)

substances

According to Article 2.1 (a) of Regulation (EC) No 1223/2009, a cosmetic product means

any substance or mixture intended to be placed in contact with the external parts of

the human body (epidermis, hair system, nails, lips and external genital organs) or with the

teeth and the mucous membranes of the oral cavity with a view exclusively or mainly to

cleaning them, perfuming them, changing their appearance, protecting them,

keeping them in good condition or correcting body odours.

“Substance” is defined by Article 2.1 (b) of this Regulation as a chemical element and its

compounds in the natural state or obtained by any manufacturing process, including any

additive necessary to preserve its stability and any impurity deriving from the process used

but excluding any solvent which may be separated without affecting the stability of the

substance or changing its composition, whereas Article 2.1 (c) defines “mixture” as a

mixture or solution composed of two or more substances.

Article 3 of the Cosmetics Regulation specifies that a cosmetic product made available on the

market shall be safe for human health when used under normal or reasonably

foreseeable conditions of use. In practice, cosmetic products have rarely been associated with

serious health hazards, which, however, does not mean that cosmetics are safe in use per

se. Particular attention is needed for long-term safety aspects, since cosmetic products may

be used extensively over a large part of the human lifespan and sensitive groups of the

population may be involved. Therefore, the safety-in-use of cosmetic products has been

established in Europe by controlling the substances, their chemical structures, toxicity

profiles, and exposure patterns.

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2. THE SCIENTIFIC COMMITTEE ON CONSUMER SAFETY, SCCS

2-1 Historical background

The Scientific Committee on Cosmetology (SCC) was established on 19 December 1977 by

Commission Decision 78/45/EEC; the purpose was to assist the European Commission in

examining the complex scientific and technical problems surrounding the drawing up and

amendment of European Union (EU) rules governing the composition, manufacturing,

packaging and labelling of cosmetic products marketed in EU countries. The Committee was

to be renewed every three years.

In 1997, the Scientific Committee on Cosmetic Products and Non-Food Products intended for

consumers (SCCNFP), was established. It was composed of independent scientists from

different fields of competence, collectively covering the widest possible range of expertise.

In 2004, the SCCNFP was replaced by the Scientific Committee on Consumer Products

(SCCP), as part of a larger-scale reorganisation of the EU Scientific Committees in the field

of consumer safety, public health and the environment.

Three scientific committees were established:

i. Scientific Committee on Consumer Products (SCCP)

ii. Scientific Committee on Health and Environmental Risks (SCHER)

iii. Scientific Committee on Emerging and Newly Identified Health Risks (SCENIHR)

The coordination between the SCCP, the SCHER and the SCENIHR was done by the Inter-

Committee Coordination Group (ICCG).

In 2008, the three above-mentioned Scientific Committees were renewed

and the SCCP's

name was changed into SCCS. In addition to the SCCS, SCENIHR and SCHER, a Pool of

scientific advisors on risk assessment was also established, with the specific task to assist the

members of the Scientific Committees in their work. In 2013, the three above-mentioned

Scientific Committees were renewed.

Finally, a new Commission Decision C (2015)5383

was adopted on 7 August 2015,

establishing two scientific committees: the (SCCS); the Scientific Committee on Health,

Environmental and Emerging Risks (SCHEER). The composition of both Committees was

renewed on April 2016, for a period of 5 years until 2021, and extended until the end of 2026

due to the Covid-crisis, which postponed the launch of the call for experts/members.

2-2 Mandate

The mission of the Scientific Committees is defined in Commission Decision C(2015)5383

which states that they shall 'provide the Commission with scientific advice and risk

assessment in the areas of public health, consumer safety, environmental risks, including,

when relevant, identification of research needs to address critical information gaps,

assessment of proposed future research actions and of research results'.

Commission Decision 2008/721/EC of 5 September 2008 setting up an advisory structure of Scientific Committees

and experts in the field of consumer safety, public health and the environment and repealing Decision 2004/210/EC.

Official Journal L 241, 10/09/2008 p.21

Commission Decision 2013/1297 of 11 March 2013 on the appointment of the members of the Scientific

Committees set up by Commission Decision 2008/721/EC.

http://ec.europa.eu/health/scientific_committees/docs/com_2013_1297_en.pdf

http://ec.europa.eu/health/scientific_committees/docs/call_2015_5383_decision_with_annexes_en.pdf

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/call_2015_5383_decision_with_annexe

s_en.pdf

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The SCCS on request of Commission services shall provide opinions on questions concerning

health and safety risks, notably chemical, biological, mechanical and other physical risks, of:

(a) non-food consumer products such as:

- cosmetic products and their ingredients, including nanomaterial, hair dyes, fragrance

ingredients;

- personal care and household products such as detergents; and toys, textiles, clothing, etc.

(b) services such as tattooing, artificial sun tanning, etc.

In addition, the Commission may request from the Committee:

- advice on any matter of particular relevance to consumer safety and public health;

- rapid advice on the state of scientific knowledge concerning specific risks in case of urgent

risks;

- the identification of research needs to address critical information gaps, to assess proposed

future research and to assess research results in relation to the subject areas covered by

its fields of competence;

- to be part of thematic networks or events with other Union bodies or scientific

organisations, in order to monitor and contribute to the development of scientific

knowledge in the fields of competence.

Also, upon its own initiative, the Committees shall draw the Commission's attention to a

specific or emerging problem falling within its remit, if it is considered to pose an actual or

potential risk to consumer safety, public health or the environment.

Finally, in agreement with the Commission, the Committees shall adopt their methodology

for performing and providing risk assessment and keep it under review to reflect all relevant

scientific factors. They shall ensure that the methodology reflect current risk assessment

practice.

The work of the SCCS can be divided in two main domains, namely matters related to

cosmetic substances and products and those related to other non-food consumer products.

Whenever cosmetic substances are concerned, the consultation of the SCCS is compulsory

whereas it is not compulsory in the domain of other non-food products.

In the preamble of Regulation (EC) No 1223/2009, different tasks for the SCCS are mentioned

in several recitals:

(28)

safety assessment of hair colorants (annex III)

(30)

providing guidance in cooperation with relevant bodies on test methodologies which take

into account specific characteristics of nanomaterials,

(32)

continuously reviewing the safety of CMR substances, so that substances clarified as CMR

2 or CMR 1A or 1B can be used in cosmetics under well-restricted conditions when such use

for CMR 1A and 1B has been found safe by the SCCS,

(34)

taking into account the exposure of vulnerable population groups,

(35)

giving opinions on the safety of use of nanomaterials in cosmetic products,

(42)

consultation by the Commission as regards the applicability of validated alternative

methods to the field of cosmetic products,

(49)

identification of substances likely to cause allergic reactions in order that their use can

be restricted and/or certain conditions can be imposed,

See Article 31 of Regulation (EC) No 1223/2009

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(61)

providing assistance to the Commission as an independent risk assessment body.

The compulsory consultation of the SCCS is taken up under:

Art. 15, 2(d) and 3 for substances classified as CMR substances

Art. 16, 4 and 5 for nanomaterials

Art. 18, 2 for animal testing methodology

Art. 20, 2 for setting criteria for product claims

Art. 27, 3 for determination whether the provisional measures taken with respect to the safe

clause are justified or not

Art. 31, 1 for amending Annexes II to VI for safety concerns

Art. 31, 2 for amending Annexes II to VI, VIII for technical and scientific progress

Art. 31, 3 for amending Annex I to ensure the safety of cosmetic products placed on the

market.

Newly introduced modifications and improvements in the current structure and working

procedures of the SCCS and the other Scientific Committee can be found in Commission

Decision C(2015)5383

of 7 August 2015.

2-3 Rules of Procedure

The Rules of Procedure

of the SCCS and SCHEER were jointly adopted by the Scientific

Committees on 28 April products. These were amended according to the Commission Decision

C(2015)5383.

In order to efficiently fulfil its extensive mandate, the SCCS sets up working groups on

particular subjects of interest. These subgroups operate independently under an appointed

chairperson (SCCS member) and consist of SCCS members complemented with external

experts (either from the Database of Experts

or via a specific call

). Working groups, for

example, deal with: Cosmetic Substances (individual substance evaluations), Methodologies

(alternative methods and Notes of Guidance), Nanomaterials and other topics according to

the needs.

The mandate on a specific substance or other issue is officially adopted by the members

during a plenary meeting (or by written procedure) and published

A Rapporteur is nominated (SCCS member or external expert). Once the participants of the

Working Groups have agreed on a final version of their opinion/scientific report(s), they

present it to the next SCCS plenary meeting where members adopt the texts. In particular

cases, an opinion may also be adopted by written procedure. The adopted preliminary

opinions, once edited, are published on the Commission’s website

for a commenting period

of a minimum of eight weeks to allow the applicant, and other stakeholders as well, to send

their comments that are subsequently considered by the SCCS and, when considered

appropriate, incorporated in a revised version of the opinion. The revised version becomes

the final opinion once adopted at the next SCCS plenary meeting (or by written procedure)

and is published on the website

, with the date of the adoption of the final text. The final

opinion replaces the preliminary opinion and informs about changes made in the first pages.

The final opinions are not subject to further comments or revision requests. SCCS is not

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/call_2015_5383_decision_with_annexe

s_en.pdf

https://ec.europa.eu/health/sites/health/files/scientific_committees/docs/rules_procedure_2016_en.pdf

http://ec.europa.eu/health/scientific_committees/experts/database/index_en.htm

http://ec.europa.eu/health/scientific_committees/open_consultation/index_en.htm

https://ec.europa.eu/health/scientific_committees/consumer_safety/requests_en

https://ec.europa.eu/health/scientific_committees/consumer_safety/opinions_en#fragment0

https://ec.europa.eu/health/scientific_committees/consumer_safety/opinions_en#fragment2

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149

responding to comment submitted outside the commenting period. Any new data should be

submitted directly to the responsible Commission unit mandating the SCCS for a new opinion.

This method of working with Working Groups not only lightens the workload of the members

of the SCCS, but equally and importantly, facilitates discussion of the individual topics with

the appropriate experts in the field of interest, thus enhancing the scientific quality of the

opinions issued.

2-4 Opinions

Before 1997, the opinions adopted by the Scientific Committee on Cosmetology at the

Commission’s request were included in EC-Reports (EUR 7297, 8634, 8794, 10305, 11080,

11139, 11303, 14208). Between 1997 and 2004, all SCCNFP opinions were published on the

Internet and can be accessed through the Committee's website

. All SCCP / SCCS opinions

can easily be located through the ingredient's substance category and the adoption date.

It must be emphasised that the SCC(NF)P / SCCS opinions and statements not only refer to

cosmetic substances included in Annexes II, III, IV, VI and VII of Council Directive

76/768/EEC or Annexes II, III, IV, V and VI of the Cosmetic Regulation (EC) No 1223/2009,

but also to a broad range of scientific issues related to the safety of cosmetic substances and

finished products.

3. COMPLYING WITH THE TESTING AND MARKETING BANS

The safety evaluation of cosmetic ingredients is exposure-driven and is historically based on

toxicological data, which were obtained by using experimental animals. The testing and

marketing bans in Regulation (EC) No 1223/2009 make the use of validated alternative

replacement methods compulsory. Guidance on how to comply can be found in:

i. Recital 50 and article 18 of the Regulation,

ii. Commission Communication on the animal testing and marketing ban and on the state

of play in relation to alternative methods in the field of cosmetics (COM/2013/135

iii. a factsheet of ECHA (2014a) and

iv. the 2017 ECHA report (ECHA 2017) on the use of alternatives to testing on animals.

I. Recital 50 of Regulation (EC) No 1223/2009 states the following: “In the safety

assessment of a cosmetic product it should be possible to take into account results

of risk assessments that have been carried out in other relevant areas. The use of

such data should be duly substantiated and justified.” The prohibitions in Article

18 of the Regulation

are triggered when the animal testing in question is done

“in order to meet the requirements of this [the Cosmetics] Regulation”. Article 18

of the Regulation (EC) No 1223/2009 creates, therefore, a relationship between

the animal testing bans and the intention to meet the requirements of this

Regulation;It is possible that animal testing needs to be conducted on ingredients

https://ec.europa.eu/health/scientific_committees/consumer_safety/opinions/sccnfp_opinions_97_04_en

COMMUNICATION on the animal testing and marketing ban and on the state of play in relation to alternative

methods in the field of cosmetics (COM(2013) 135 final).

Article 18 of Regulation (EC) No 1223/2009 contains four prohibitions, two relating to the performance of animal

testing (on finished cosmetic products and on ingredients of cosmetic products) and two relating to the placing on

the market of cosmetic products (where the final formulation or an ingredient of a cosmetic product has been the

subject of animal testing). However, an option for derogation from the animal testing ban is foreseen in Article 18,

No 2, paragraph six “In exceptional circumstances, where serious concerns arise as regards the safety of an existing

cosmetic ingredient, a Member State may request the Commission to grant a derogation from paragraph 1. The

request shall contain an evaluation of the situation and indicate the measures necessary. On this basis, the

Commission may, after consulting the SCCS and by means of a reasoned decision, authorise the derogation. That

authorisation shall lay down the conditions associated with this derogation in terms of specific objectives, duration

and reporting of the results”.

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150

to be used in a cosmetic product for the purpose of complying with other regulatory

framework (e.g., food, medicines, biocides).

II. In this respect, Commission Communication COM/2013/135 further elucidates: “If

animal testing was involved and took place after the 2013 deadline, the product

information file should allow verification on whether the testing was carried out in

order to meet the requirements of the Regulation or for other purposes. To this

end the file should contain documentation on any use of the substance in products

other than cosmetic products (product examples, market data etc.), as well as

documentation on compliance with other regulatory frameworks (e.g. REACH or

other legal frameworks) and a justification of the need for the animal testing under

that other framework (e.g. testing proposal under REACH)” . As regards the use

of data from animal testing conducted to ensure compliance with non-cosmetics

related legislative frameworks, two different scenarios can occur:

a. With respect to ingredients that are equally in use in other consumer and

industrial products, such as in pharmaceuticals, detergents and food, animal

testing may be necessary to ensure compliance with the legal frameworks

applicable to these products. In this case, the Commission considers that “the

resulting animal testing data should not trigger the marketing ban and could

subsequently be relied on in the cosmetics safety assessment. Reliance on such

data is subject to its relevance for the cosmetics safety assessment and its

compliance with data quality requirements”. However, the Commission

Communication COM/2013/135 also adds that it is “for Member States to assess

and decide whether such testing for compliance with other frameworks is

considered to be falling in the scope of the 2013 marketing ban”;

b. Conversely, animal testing conducted on ingredients that have been specifically

developed for cosmetic purposes and are exclusively used in cosmetic products

would in the Commission's view always be assumed to be carried out in order to

meet the requirements of the Regulation (EC) No 1223/2009

, i.e. would always

be assumed to fall under the scope of the Article 18 ban. It would not be possible,

therefore, to use the results of such animal testing to prove safety of cosmetic

ingredients.

III. With respect, in particular, to the interaction between REACH requirements and

animal testing, ECHA published a factsheet

aimed at clarifying the practical

meaning and implications of the Commission Communication COM/2013/135 in

the context of REACH. The interface between REACH and the Regulation (EC) No

1223/2009 has been illustrated in a scheme, see Appendix 4. It has to be noted

that animal testing under REACH is not restricted, if: a) this testing is required for

environmental endpoints; or b) the substance is also registered for non-cosmetic

uses. Even if a substance is registered exclusively for cosmetic use, the animal

testing requirements continue to apply to tests needed to assess the risks from

exposure to workers in the Chemical Safety Assessment (ECHA, 2014a

). For the

first time, on 18 August 2020, the Board of Appeal (BoA) of ECHA took two

“Testing carried out for cosmetics relevant endpoints on ingredients that have been specifically developed for

cosmetic purposes and are exclusively used in cosmetic products would in the Commission's view always be

assumed to be carried out 'in order to meet the requirements of this Directive/Regulation'” (Commission

Communication COM/2013/135, Page 8).

https://echa.europa.eu/documents/10162/13628/reach_cosmetics_factsheet_en.pdf

“Workers” in this context are to be understood as persons who are actively involved in a particular activity of a

production or manufacturing site where they may be exposed directly or indirectly to chemical substances. On the

other hand, professional users who use the cosmetic products as part of their professional activity (e.g. hairdressers)

and consumers shall not be considered as “workers”. In Regulation (EC) No 1223/2009 the term ‘end user’ means

either a consumer or professional using the cosmetic product (Article 2, Definitions 1.

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151

compliance check decisions

on registration dossiers (for homosalate and 2-

ethylhexyl salicylate, both UV filters used exclusively in cosmetics) (ECHA 2020a

and 2020b) where it confirmed that, according to scientific evidence, ECHA may

conclude that studies on vertebrate animals must be provided by the applicant to

comply with REACH, even if the substance is used exclusively as an ingredient in

cosmetics. This said, the considerations under point II above would apply, meaning

that, as regards ingredients that have been specifically developed for cosmetic

purposes and are exclusively used in cosmetic products, the results of a study on

vertebrate animals required under REACH could not be relied upon in the cosmetic

product safety report in order to demonstrate the safety for end users, as these

would fall under the Article 18 ban.

However, such results will be available to the authorities for scrutiny in the cosmetic product

information file under Article 11 of the Regulation (EC) No 1223/2009 and might call into

question the safety of cosmetic products containing a registered substance, contradicting the

cosmetic product safety report. In this case, as mentioned by the ECHA BoA in case A-010-

2018

“if the safety of cosmetic products containing the substance can no longer be

established, then it is possible that cosmetic products containing the substance in question

as an ingredient can no longer be placed on the market” (paragraph 112). The need to take

into account the consequence of the results of that study would be justified under Article 3 of

the Regulation (EC) No 1223/2009, which provides that a cosmetic product made available

on the market must be safe for human health when used under normal or reasonably

foreseeable conditions of use.

IV. Additional information regarding the REACH legislation in the context of alternative

methods can be found in the three reports on “The Use of Alternatives to Testing

on Animals for the REACH Regulation”, in the 3

report under Article 117(3),

available online

(https://echa.europa.eu/documents/10162/13639/alternatives_test_animals_20

17_en.pdf)

The question of the interpretation of the animal testing ban as regards animal testing

performed in third countries to comply with the cosmetics legislation of a third country was

referred to the European Court of Justice in case C-592/14

. The Court concluded that: ''the

results of animal tests, carried out outside the European Union in order to market cosmetic

products in third countries, the results of which are used to prove the safety of those products

for the purpose of their being placed on the EU market, must be regarded as having been

carried out ‘in order to meet the requirements [of that regulation]’ […]. ''Article 18(1)(b) of

Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November

2009 on cosmetic products must be interpreted as meaning that it may prohibit the placing

on the European Union market of cosmetic products containing some ingredients that have

been tested on animals outside the European Union, in order to market cosmetic products in

third countries, if the resulting data is used to prove the safety of those products for the

purposes of placing them on the EU market''.

The information provided in the NoG relates to the assessment of cosmetic ingredients from

a general chemical safety point of view. However, safety assessment of chemical substances

in certain physicochemical forms may need additional specific considerations, for example,

the use of nanomaterials in cosmetics (SCCS/1611/19).

https://echa.europa.eu/about-us/who-we-are/board-of-appeal

https://echa.europa.eu/documents/10162/23010712/a-010-2018_decision_en.pdf/46612b84-29af-29ea-9192-

b2506f33c8ce

Judgment of 21 September 2016, European Federation for Cosmetic Ingredients, C-592/14, ECLI:EU:C:2016:703.

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APPENDIX 2: LISTS OF SUBSTANCES

1. INTRODUCTION

Regulated cosmetic substances can be found as Annexes II, III, IV, V and VI to Regulation

(EC) No 1223/2009. These annexes lay down clear limitations and requirements for the

cosmetic substances concerned.

Another important list of cosmetic substances is the INCI (International Nomenclature

Cosmetic Ingredient) inventory (96/335/EC) or CIN (2009/1223/EC), identifying a large

number of substances with their possible function(s) in finished cosmetic products and with

the nomenclature that needs to be used on the label of finished cosmetic products. DG GROW

(Directorate-General for Internal Market, Industry, Entrepreneurship and SMEs) has built up

a free to use database of cosmetic substances called CosIng,

http://ec.europa.eu/consumers/cosmetics/cosing

(Cosmetic ingredients) which combines INCI names and synonyms of the listed substances

with useful regulatory information. CosIng database is regularly updated with information on

new cosmetics ingredients. The information contained in CosIng is indicative and does

not have any legal value.

Finally, this section briefly mentions Annex I to the Dangerous Substances Legislation

(67/548/EEC), since the "7

Amendment" of Directive 76/768/EEC (2003/15/EC) and the

Recast (2009/1223/EC) directly refer to that list when excluding CMR Cat.1 & Cat.2 chemicals

from cosmetic use (see 3-6.6). With the European Regulation on classification and labelling

(2008/1272/EC), however, Annex I to Dir. 67/548/EEC now needs to be referred to as ‘Part

3 of Annex VI to Regulation (EC) No 1272/2008’, in which all existing European classifications

are converted into new harmonised classifications using the new criteria.

It must be emphasised that none of the above lists reflects the complete set of substances

used in cosmetic products.

2. ANNEXES II, III, IV, V AND VI TO THE COSMETIC PRODUCTS REGULATION

The Cosmetic Products Regulation defines Annexes II, III, IV V and VI, which have been

described in Section 3-1.

3. INVENTORY OF SUBSTANCES USED IN COSMETIC PRODUCTS

Article 33 of Regulation (EC) No 1223/2009 states that the Commission shall compile and

update a glossary of common ingredient names (CINs) employed in cosmetic products

(2003/1223/2009).

On 8 May 1996, the European Commission established an Inventory and a common

nomenclature of the substances employed in cosmetic products (96/335/EC, part of which

amended by 2006/257/EC). This list was subdivided into 2 sections:

Section I: Inventory of ingredients employed in cosmetic products

Section II: Perfume and aromatic raw materials

The Inventory is indicative and does not constitute a list of substances authorised for use in

cosmetic products. If an INCI name is available, it is to be used on the packaging and

labelling, but the absence of an INCI name on the Inventory does not automatically exclude

the use of the substance under consideration.

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153

An entry in the Inventory provides identification of that particular substance through the

following parameters:

- Common name: INCI; but botanicals get their systemic (Linné) Latin names and

colourants a colour index (CI) number

- Chemical name

- Chemical Abstract Service (CAS) number

- European Pharmacopoeia (Ph. Eur.) name

- International Non-proprietary Name (INN) name, recommended by WHO

- International Union of Pure and Applied Chemistry (IUPAC) name

- EC number, meaning either:

European Inventory of Existing commercial Chemical Substances (EINECS) number

(format 2xx-xxx-x)

European List of Notified Chemical Substances (ELINCS) number (format 4xx-xxx-x)

No Longer Polymer (NLP) number (format 5xx-xxx-x)

EC Number appointed under REACH procedure (format 6xx-xxx-x or 7xx-xxx-x)

In 1998 the European Commission issued a Mandate (DG24/XXIV/1891/98), indicating that

the SCCNFP shall act as a resource of scientific expertise to the European Commission, in

terms of advising on the:

- medical and professional expectations and requirements of the Inventory,

- scientific accuracy and validity of proposed entries,

- outstanding needs of the existing text /proposed improvements in subsequent updates.

After collaboration with the JRC (Joint Research Centre) of the Commission, the experts from

European industry and Colipa (the European Cosmetic Toiletry and Perfumery Association;

now called Cosmetics Europe), the SCCNFP issued a Status Report on the Inventory

(SCCNFP/0098/99). In this report, 6 priorities were identified for a first update of the INCI

list:

1) To accomplish the principle: each INCI name should refer to only one specific

substance.

2) To correct the INCI names of Ethylhexyl derivatives and to adopt a final decision on

Ampho-derivatives.

3) To identify botanical entries with greater transparency.

4) To solve problems on chemical identification associated to polymers.

5) To solve the problem of hair dyes/cosmetic colourants with respect to Colour Index (CI)

identification and restrictions.

6) To improve the description of the functions of the substances.

Having taken into account this list of priorities, the SCCNFP published in June 2000 "The 1

Revision and Update of Section I of the Inventory of ingredients employed in cosmetics"

(SCCNFP/0299/00). This update contains many improvements to the original edition of

Section I, including 1466 new and 843 modified INCI names, as well as a number of necessary

recommendations for updating the inventory in the future.

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154

In October 2000, "The 1

Update of the Inventory of ingredients employed in cosmetic

products: Section II: Perfume and aromatic raw materials" was issued (SCCNFP/0389/00).

Again, many improvements were introduced (e.g. 650 new entries of botanicals) and

recommendations for future updates were added.

In 2006, Commission Decision 2006/257/EC established the most recent official list

containing the common nomenclature of ingredients employed in cosmetic products

(2006/257/EC).

From 11 July 2013 on, the INCI list will be replaced by the so-called "Common Ingredients

glossary" (2009/1223/EC). The new glossary will contain the harmonised names of

approximately 26.000 cosmetic substances.

4. COSING - EC INFORMATION ON COSMETIC SUBSTANCES

The CosIng database

is a publicly available information database in two parts, linked together

whenever possible. One part aims at containing all the regulations introduced by the Cosmetic

Directive/Regulation. This part contains the historical data since the beginning of the

Cosmetics Directive in 1976. The scientific opinions, which are the basis for many of the

authorised substances or the restrictions of the substances in the Annexes, are linked to the

regulated substances. Each substance is provided with the chemical name, INN name or

IUPAC-name, CAS- and EC number, Annex and entry number and the conditions and warnings

for its use.

The other part of the database contains the EU-inventory, which is a list of assigned INCI-

names to substances offered for sale to the cosmetic industry. In addition to the INCI-name,

if possible the CAS- and EC number, chemical name or its description is added, together with

the function in the cosmetic products and finally any restrictions imposed by the Cosmetics

Directive.

Every possible link between the 2 parts has been established.

5. PART 3 OF ANNEX VI TO REGULATION (EC) NO 1272/2008

Part 3 of Annex VI to Regulation (EC) No 1272/2008 provides the harmonised European

classification of a large number of dangerous substances according to the principles laid down

in Annex I to that same Regulation (2008/1272/EC). Annex VI Part 3 previously was Annex I

to Directive 67/548/EEC, which was repealed in December 2010. The European harmonised

classification Annex is updated on a regular basis and contains a large number of chemicals

that can be found in the composition of cosmetic products. It is useful to check the

harmonised classification of a compound of interest, but it is of particular importance with

regard to Art. 15 of the Cosmetic Products, which states (2009/1223/EC):

The use in cosmetic products of substances classified as carcinogenic, germ cell mutagenic

or toxic for reproduction, of category 1A, 1B and 2, under part 3 of Annex VI to Regulation

(EC) No 1272/2008 shall be prohibited ... A substance classified in category 2 may be used

in cosmetics if the substance has been evaluated by the Scientific Committee on Consumer

Safety (SCCS) and found acceptable for use in cosmetic products.

http://ec.europa.eu/consumers/cosmetics/cosing/ Consulted December 2020.

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155

APPENDIX 3: STANDARD FORMAT OF THE OPINIONS

Scientific Committee on Consumer Safety

SCCS

OPINION ON

……………………………………………

The SCCS adopted this document

at its plenary meeting/by written procedure on xx

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156

ACKNOWLEDGMENTS

Members of the Working Group are acknowledged for their valuable contribution to this

Opinion. The members of the Working Group are:

The SCCS members:

………………………………………

The SCHEER members (if applicable):

………………………………………

External experts (if applicable):

………………………………………

The additional contribution of the following experts is gratefully acknowledged (if applicable):

XXXXXX

All Declarations of Working Group members are available on the following webpage:

https://ec.europa.eu/transparency/regexpert/index.cfm

If relevant: This Opinion has been subject to a commenting period of XXX weeks (from ……

to ……) after its initial publication.

There were comments received and the final version of the Opinion includes information on

XXXX (section concerned)….compared to the preliminary one.

There were changes/no change in the conclusions.

OR - There were no comments received and the final version of the opinion remained

unchanged compared to the preliminary one.

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157

1. ABSTRACT

Text from the rapporteur

The SCCS concludes the following:

Response

etc

Keywords: SCCS, scientific opinion, INCI name, type of product, Regulation 1223/2009,

CAS ………, EC …………………

Opinion to be cited as: SCCS (Scientific Committee on Consumer Safety), Opinion on INCI

name - Submission …., preliminary version of (date), final version of (date),

SCCS/…../XX……………………………………………………………………………………………

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158

About the Scientific Committees

Two independent non-food Scientific Committees provide the Commission with the scientific

advice it needs when preparing policy and proposals relating to consumer safety, public health

and the environment. The Committees also draw the Commission's attention to the new or

emerging problems which may pose an actual or potential threat.

These Committees are: the Scientific Committee on Consumer Safety (SCCS) and the

Scientific Committee on Health, Environmental and Emerging Risks (SCHEER) and they are

made up of scientists appointed in their personal capacity.

In addition, the Commission relies upon the work of the European Food Safety Authority

(EFSA), the European Medicines Agency (EMA), the European Centre for Disease prevention

and Control (ECDC) and the European Chemicals Agency (ECHA).

SCCS

The Committee shall provide Opinions on questions concerning all types of health and safety

risks (notably chemical, biological, mechanical and other physical risks) of non-food consumer

products (for example: cosmetic products and their ingredients, toys, textiles, clothing,

personal care and household products such as detergents, etc.) and services (for example:

tattooing, artificial sun tanning, etc.).

Scientific Committee members

Ulrike Bernauer, Laurent Bodin, Qasim Chaudhry, Pieter Jan Coenraads, Maria Dusinska,

Janine Ezendam, Eric Gaffet, Corrado Lodovico Galli, Berit Granum, Eirini Panteri, Vera

Rogiers, Christophe Rousselle, Maciej Stepnik, Tamara Vanhaecke, Susan Wijhoven

Contact

European Commission

Health and Food Safety

Directorate C: Public Health

Unit C2: Health information and integration in all policies

L-2920 Luxembourg

SANTE-C2-SCCS@ec.europa.eu

European Union, 20XX

ISSN ISBN

Doi: ND-

The Opinions of the Scientific Committees present the views of the independent scientists

who are members of the committees. They do not necessarily reflect the views of the

European Commission. The Opinions are published by the European Commission in their

original language only.

http://ec.europa.eu/health/scientific_committees/consumer_safety/index_en.htm

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159

2. MANDATE FROM THE EUROPEAN COMMISSION

Background

Terms of reference

Additional information

(If appropriate)

This chapter could provide additional background information relevant to the assessment

(e.g. previous Opinions or other assessments issued by other bodies/organisations).

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160

3. OPINION

3.1 CHEMICAL AND PHYSICAL SPECIFICATIONS

3.1.1 Chemical identity

3.1.1.1 Primary name and/or INCI name

3.1.1.2 Chemical names

3.1.1.3 Trade names and abbreviations

3.1.1.4 CAS / EC number

3.1.1.5 Structural formula

3.1.1.6 Empirical formula

3.1.2 Physical form

3.1.3 Molecular weight

3.1.4 Purity, composition and substance codes

3.1.5 Impurities / accompanying contaminants

3.1.6 Solubility

3.1.7 Partition coefficient (Log P

)

3.1.8 Additional physical and chemical specifications

Where relevant:

- organoleptic properties (colour, odour, taste if relevant)

- melting point

- boiling point

- flash point

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161

- vapour pressure

- density

- viscosity

- pKa

- pH

- refractive index

- UV/visible light absorption spectrum

3.1.9 Homogeneity and Stability

3.2 EXPOSURE ASSESSMENT & TOXICOKINETICS

3.2.1 Function and uses

3.2.2 Dermal / percutaneous absorption

3.2.3 Other studies on toxicokinetics

3.2.4 Calculation of SED/LED

3.3 TOXICOLOGICAL EVALUATION

3.3.1. Irritation and corrosivity

3.3.1.1 Skin irritation

3.3.1.2 Mucous membrane irritation / eye irritation

3.3.2 Skin sensitisation

3.3.3 Acute toxicity

3.3.3.1 Acute oral toxicity

3.3.3.2 Acute dermal toxicity

3.3.3.3 Acute inhalation toxicity

3.3.4 Repeated dose toxicity

3.3.4.1 Repeated dose (28 days) oral / dermal / inhalation toxicity

3.3.4.2 Sub-chronic (90 days) oral / dermal / inhalation toxicity

3.3.4.3 Chronic (> 12 months) toxicity

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162

3.3.5 Reproductive toxicity

3.3.5.1 Fertility and reproduction toxicity

3.3.5.2 Developmental Toxicity

3.3.6 Mutagenicity / genotoxicity

3.3.6.1 Mutagenicity / genotoxicity in vitro

3.3.6.2 Mutagenicity / genotoxicity in vivo

3.3.7 Carcinogenicity

3.3.8 Photo-induced toxicity

3.3.8.1 Phototoxicity / photo-irritation and photosensitisation

3.3.8.2 Photomutagenicity / photoclastogenicity

3.3.9 Human data

3.3.10 Special investigations

3.4 SAFETY EVALUATION (INCLUDING CALCULATION OF THE MoS)

3.5 DISCUSSION

Physicochemical properties

Exposure & Toxicokinetics

Toxicological Evaluation

Irritation and corrosivity

Skin sensitisation

Acute toxicity

Repeated dose toxicity

Reproductive toxicity

Mutagenicity / genotoxicity

Carcinogenicity

Photo-induced toxicity

Human data

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163

Special investigation

4. CONCLUSION

Response

etc

5. MINORITY OPINION

6. REFERENCES

7. GLOSSARY OF TERMS

See SCCS/1628/21, 11th Revision of the SCCS Notes of Guidance for the Testing of Cosmetic

Ingredients and their Safety Evaluation – from page 181

8. LIST OF ABBREVIATIONS

See SCCS/1628/21, 11th Revision of the SCCS Notes of Guidance for the Testing of Cosmetic

Ingredients and their Safety Evaluation – from page 181

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164

APPENDIX 4: ANIMAL TESTING: INTERFACE BETWEEN REACH AND

COSMETICS REGULATIONS

Reference: Interface between REACH and Cosmetics regulations (ECHA, 2014a)

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165

APPENDIX 5: CMR GUIDANCE ON SAFE USE OF CMR SUBSTANCES IN

COSMETIC PRODUCTS

GUIDANCE ON A HARMONISED APPROACH TO THE DEVELOPMENT AND USE OF

OVERALL EXPOSURE ESTIMATES IN ASSESSING THE SAFE USE OF CMR SUBSTANCES

IN COSMETIC PRODUCTS

I. Background

1. Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30

November 2009 on cosmetic products

(Cosmetics Regulation) contains in its Article 15

provisions on the use in cosmetic products of substances classified as carcinogenic, mutagenic

or toxic for reproduction (CMR substances) under Part 3 of Annex VI to Regulation (EC)

1272/2008

. These provisions apply from 1 December 2010.

2. As a general rule, the substances classified as CMR substances of category 1A, 1B and 2

under Part 3 of Annex VI to Regulation (EC) 1272/2008 are prohibited for use in cosmetic

products.

3. However, exceptions to this rule are foreseen by the Cosmetics Regulation. Indeed, a

substance classified as a CMR substance of category 2 may be used in cosmetic products

where the substance has been evaluated by the Scientific Committee on Consumer Safety

(SCCS) and found safe for use in cosmetic products on the basis of the data submitted.

4. Also, CMR substances of category 1A or 1B may be used in cosmetic products by way of

exception where, subsequent to their classification as CMR substances of category 1A or 1B

under Part 3 of Annex VI to Regulation (EC) No 1272/2008, all of the following conditions are

fulfilled:

(a) they comply with the food safety requirements as defined in Regulation (EC) No 178/2002

of the European Parliament and the Council of 28 January 2002 laying down the general

principles and requirements of food law, establishing the European Food Safety Authority

and laying down procedures in matters of food safety

;

(b) there are no suitable alternative substances available, as documented in an analysis of

alternatives;

exposure; and

(d) they have been evaluated and found safe by the SCCS for use in cosmetic products, in

particular in view of exposure to these products and taking into consideration the overall

exposure from other sources, taking particular account of vulnerable population

subgroups.

II. Scope and objectives

5. Article 15, paragraph 3 of the Cosmetics Regulation foresees that the Commission shall

ensure that appropriate guidance is developed with the aim of enabling a harmonised

approach to the development and use of overall exposure estimates in assessing the safe use

of CMR substances.

OJ L 342, 22.12.2009, p. 59.

OJ L 353, 31.12.2008, p. 1.

OJ L 31, 1.2.2002, p. 1.

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166

6. To authorise the use of CMR substances of category 1A or 1B in cosmetic products, one of

the conditions to be fulfilled is that they have been evaluated and found safe by the SCCS for

use in cosmetic products, in particular in view of exposure to cosmetics products and taking

into consideration the overall exposure from other sources and vulnerable population

subgroups.

7. On a case-by-case basis and at the request of the SCCS, it may also be necessary to

perform an overall exposure from other sources for CMR 2 substances. Therefore, the

procedure developed below for the overall exposure assessment of CMR 1A and 1 B

substances should, where necessary, also apply to CMR 2 substances (condition (d) only)

8. Appropriate consultations with the SCCS and other relevant stakeholders have been carried

out in order to develop this guidance. In addition, administrative agreements have been

established with relevant EU Agencies - European Chemicals Agency (ECHA), European Food

Safety Authority (EFSA), European Medicines Agency (EMA) - to ensure the appropriate

exchange of data between them and the SCCS Secretariat.

III. Procedure

9. The aim of this guidance is to outline the mechanisms necessary for ensuring the

generation and the exchange of the appropriate data for the assessment by the SCCS of the

overall exposure to a CMR 1A or 1B substance stemming from other sources than cosmetics

(such as food, biocides, etc.).

10. When a substance of interest for the industry is indicated in the Registry of Intentions for

the purpose of its harmonised classification as CMR substance under Part 3 of Annex VI to

Regulation (EC) No 1272/2008, it is for the industry to inform the Commission in due time of

its intention to defend a substance under discussion to allow that any possible derogation

measure is adopted by the Commission within the timeframe defined by Article 15 of the

Cosmetics Regulation 1223/2009.

11. The Commission responsible Services should inform the SCCS that the industry intends to

defend the substance. They should also inform the Member States of this intention, so that

any relevant data available in public or state laboratories, or elsewhere, may be considered

for the scientific assessment. In parallel, they may also organise a call for scientific data from

anyone holding or being aware of further relevant information, in order to gather additional

scientific data.

12. It is the industry's responsibility to demonstrate that the first three conditions (a), (b)

and (c) for derogation laid down in Article 15 paragraph 2 of Cosmetics Regulation are

fulfilled. For justifying compliance with each of the above conditions, the industry should

submit appropriate dossiers for examination by the Commission responsible Services.

13. The Commission responsible Services should verify the compliance with the food safety

requirements, where necessary by consulting the EFSA, and verify the absence of suitable

alternative substances and the fact that the application is limited for a particular use of the

product category with a known exposure, where necessary by consulting the Standing

Committee on Cosmetic Products.

14. Subsequently, the procedure for the exchanges of data between the relevant entities can

be started as regards to the overall exposure assessment by the SCCS (condition d). Requests

for data sharing with the relevant EU Agencies (ECHA, EFSA and EMA

) should be initiated

and managed by the SCCS Secretariat. On a case by case basis, the Commission responsible

Services can, where relevant, ask for data to Member States or third countries.

The need to consult EMA will be checked by the Commission on a case by case basis.

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167

15. The "Declaration of Commitment by the Commission with respect to security aspects for

ECHA's information systems" has been signed by the responsible Commission Services

and

sets up the conditions under which exchange of confidential data from REACH dossiers can

be ensured with ECHA.

16. Upon request by the SCCS Secretariat, the Commission responsible Services should grant

access to relevant data in REACH registration dossiers to a designated SCCS expert who

adheres to the security rules for users of ECHA's Information System.

17. The extraction of relevant data from REACH dossiers and their processing to establish

aggregated exposure levels should be completed by the designated SCCS expert within the

secure room of the Commission responsible Services and in accordance with all applicable

security rules. In case an evaluation of the CMR substance has already been completed under

REACH, exposure levels that have been established can also be used straightaway where

appropriate.

18. The EFSA should be consulted by the SCCS Secretariat to provide, if available, data or

estimates on exposure from food and other relevant sources.

19. Additionally, the EMA could be consulted by the SCCS Secretariat on a case-by-case basis

on exposure from substances used as pharmaceuticals.

20. The applicant should include in their submission all of the exposure information that they

have. In addition to the exposure information gathered as mentioned above, e.g., exchange

of data with the Agencies, public call for information, consultation with Member States, the

SCCS will consider the exposure information provided by the applicant.

21. It is necessary that the exchange of data takes place in a smooth and timely manner as,

for CMR 1A and 1B substances, the measure necessary for the derogation must be adopted

by the Commission within 15 months following the adoption of the classification as CMR

substance.

22. The SCCS, once it has received the scientific data from ECHA, EFSA, EMA and has taken

into consideration the data submitted by the industry and other available sources (such as

information gathered from Member States or following public consultation), shall assess the

specific CMR substance(s) for safety of use in cosmetic products taking into account the

overall exposure from other sources and vulnerable population groups within a timescale of

at least six months for finalising their Opinion after an adequate submission and a complete

set of exposure data is received.

23. It should be noted that, where the work of other scientific/regulatory bodies contains

information on exposure to humans via the environment, this may have been incorporated in

their overall estimates of exposure. However, Cosmetic Regulation (EC) No 1223/2009 only

covers the aspects of safety to human health. As indicated in recital 5 of that Regulation, the

environmental concerns that substances used in cosmetic products may raise are considered

through the application of Regulation (EC) No 1907/2006 (REACH)

24. As regards the scientific risk assessment of CMR substances of categories 1A and 1B used

in cosmetics, the SCCS will determine the most appropriate methodology for their safety

evaluation based on the best scientific knowledge and taking into account the exposure from

the specific uses in cosmetic products and the overall exposure from other sources.

DG ENTR and DG ENV co-managed the REACH legislation.

OJ L 396, 30.12.2006, p. 1.

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168

25. In order to provide transparency on the applied methodology and guidance to the industry,

the SCCS should develop and incorporate this methodology within the next revision of its

"Notes of Guidance for the testing of cosmetic substances and their safety evaluation"

IV. Final observations

26. This document is only meant to provide guidance for a harmonised approach to the

development and use of overall exposure estimates in assessing the safe use of CMR

substances in cosmetic products and it is by no means binding.

27. The SCCS evaluation will not automatically trigger action under any legislation other than

the Cosmetics legislation. The SCCS conclusions will be publicly available.

28. This document may be revised in the future in light of further scientific developments.

SCCS/1564/15 of 29 September 2015, revised on 16 March 2016.

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169

APPENDIX 6: REQUIREMENTS FOR THE CERTIFICATE OF ANALYSIS

FOR A COSMETIC INGREDIENT

The Certificate of Analysis for a cosmetic ingredient should include:

1. The name and address of the laboratory performing the tests.

2. The registration number of the certificate of analysis.

3. The name, description and number of the batch for which the certificate is issued, the

date of manufacture, and the expiry date.

4. The date on which the batch for which the certificate is issued was received.

5. A reference to the test procedure used, including the acceptance criteria (limits).

6. The results of all tests performed on the batch for which the certificate is issued (in

numerical form, where applicable) and a comparison with the established acceptance

criteria (limits), including information on Appearance, Identity (IR, NMR, MS), Purity,

Solubility, Impurities (% content), Heavy metals.

7. Any additional test results obtained on samples from the batch as part of a periodic

statistically based testing program

8. A statement indicating whether the results were found to comply with the

requirements.

9. The date(s) on which the test(s) was (were) performed.

10. The signature of the head of the laboratory or an authorised person.

11. The name, address, and telephone and fax numbers of the original manufacturer. If

supplied by repackers or traders, the certificate should show the name, address, and

telephone and fax numbers of the repacker/trader and a reference to the original

manufacturer.

12. A statement of the expected conditions of shipping, packaging, storage and

distribution, deviation from which would invalidate the certificate.

13. A copy of the certificate generated by the original manufacturer, if the sample is

supplied by a repacker or trader.

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170

APPENDIX 7: DETAILED EXPOSURE DATA FOR COSMETIC PRODUCTS

During the last years, exposure data for several cosmetic product categories became available

in the open literature. This can be useful for safety assessors and safety agencies when in

some particular cases refinement of risk assessment is necessary to show product or

ingredients safety. In Table A.7 a literature overview is provided of recent cosmetic product

consumer exposure data (e.g. different categories of cosmetics with frequency of use, amount

per application, amount per day) which are focused on consumers from one or more particular

countries. In a number of cases, data are shown stratified by age and/or gender, and for

different cosmetic formulations.

Table A.7: literature overview (2015-2020) of specific cosmetic consumer exposure data and

assessments

References

Country(ies)

Product categories

Additional information

Husoy et al., 2020

Norway

cosmetic products and

toothpaste

Adults, both genders

Gomez-Berrada et

al., 2018a

France

toothpaste

adults and children;

both genders

Gomez-Berrada et

al., 2018b

France

sunscreens

adults and children; both

genders under real-life

conditions

Bernard et al., 2018

France

face and oral care

cosmetic products

probabilistic exposure

assessment; both genders;

different age groups

Gomez-Berrada et

al., 2017

France/ (1 city: Rennes)

cosmetic products

children under 2 years

consumption; exposure

assessment

Ficheux and Roudot

2017

France

cosmetic products

general population; both

genders; different age groups

Dornic et al., 2017a

France

perfumes in cosmetic

products

adults and children

Dornic et al., 2017b

France

default values for skin surface

area

Dornic et al., 2017c

France

cosmetic products

exposure data;

both genders, different age

groups

Lee et al., 2017

Korea

baby care products

children 0-3 years

Garcia-Hidalgo et al.,

2017

Swiss

personal care products

use patterns both genders;

different age groups

Rieder et al., 2017

cosmetic ingredient

case of tea tree oil

Strittholt et al., 2016

toothpaste

in children (2-7yrs)

Bernard et al., 2016a

France

hair dye products

both genders

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171

use patterns; exposure

assessment

Ficheux et al., 2016a

France

different cosmetic products

children (0-3yrs)

Ficheux et al., 2016b

France

different hair cosmetic

products

both genders

Ficheux et al., 2016c

Ficheux et al.,2019

France

different cosmetic products

consumption amounts;

different age groups; both

genders

probabilistic aggregate exposure

for babies, children; both

genders

Dey et al., 2016a

USA, Germany, UK

baby wipes

lotion transfer via baby wipes

Dey et al., 2016b

world

exposure factor of disposable

diapers

Comiskey et al.,

2015

EU, USA

fragrance ingredients

probabilistic aggregate exposure

Manová et al., 2015

Swiss, Germany

UV filter

ethylhexylmethoxy-

cinnamate

probabilistic aggregate exposure

Tozer et al., 2015

USA

Zn pyrithione in rinse-off

personal cleansing

products

probabilistic aggregate exposure

Dudzina et al., 2015

siloxane D5

probabilistic aggregate exposure

(PACEM)

Nijkamp et al., 2015

fragrance geraniol in

personal care products

probabilistic aggregate exposure

Safford et al., 2015

fragrance ingredients in

cosmetic and personal care

products

probabilistic aggregate exposure

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172

APPENDIX 8: KEY CHARACTERISTICS OF CARCINOGENS

In the overall WoE assessment of a cosmetic ingredient, 10 key characteristics commonly

exhibited by established human carcinogens can be taken into account (Smith et al., 2019,

Al-Zougholl, 2019). High-throughput assay systems, such as the US EPA’s ToxCast program

(Chiu et al., 2018), which provide in vitro mechanistic data on several of the key

characteristics, may be helpful.

Table A.8: Key characteristics of carcinogens (based on: Smith et al., 2019 and Al-Zougholl

et al., 2019); AhR, aryl hydrocarbon receptor; ER, estrogen receptor; PPAR, peroxisome proliferator–

activated receptor.

Characteristic

Description

1. Is electrophilic or can be

metabolically activated to

electrophiles

Formation of protein adducts indicates the presence of reactive

chemicals, which are sometimes also considered as indirect

indicators/predictors of DNA damage (see characteristic 2, below)

Requires biotransformation (metabolic activation) to produce reactive

metabolites, e.g. alkylating agents, epoxide metabolites, aryl-nitrenium

ion

Evidence for ADME of the agent affecting its carcinogenicity

2. Is genotoxic

Direct evidence of DNA damage – this category includes nuclear and

mitochondrial DNA damage (in vitro or in vivo): DNA adducts, DNA

strand breaks (single- and/or double-strand breaks), DNA–protein cross-

links, DNA–DNA cross-links.

Indirect indicators or biomarkers of DNA damage (in vitro or in vivo).

Disruption or breakages of chromosomes leading to sections of the

chromosome being deleted, added, or rearranged.

Reversions and forward mutations in microorganisms or mammalian

cells. Mutations affecting oncogenes, tumour-suppressor genes, and

other genes involved in cell cycle control.

3. Alters DNA repair or

causes genomic instability

Effects on key DNA-repair mechanisms such as base-excision repair

(BER) and nucleotide-excision repair (NER). Inherited abnormalities in

DNA-repair function lead to enhanced cancer susceptibility.

4. Induces epigenetic

alterations

Stable, long-term alterations in the transcriptional potential of a cell.

These effects can be caused by factors such as altered methylation of

DNA, micro-RNA expression, and changes in chromatin and histone

structure.

5. Induces oxidative stress

Disturbance in the balance between the production of reactive oxygen

species (free radicals) and antioxidant defenses within a cell.

6. Induces chronic

inflammation

Chronic inflammation and/or irritation leading to oxidative DNA damage.

7. Is immunosuppressive

Measures of altered function of the immune system that may lead to

increased cancer risk (e.g. HIV-related effects).

8. Modulates receptor-

mediated effects

Interference with cell-signaling pathways leading to expression of

carcinogenic trait/phenotype in the cell, e.g. facilitating cell invasion or

induction of genes for inflammatory mediators, oncogenes

Interference with the synthesis, secretion, transport, binding, action, or

elimination of natural hormones in the body. External agents can

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173

interfere with the synthesis, secretion, transport, binding, action, or

elimination of natural hormones in the body.

9. Causes immortalization

A. Oncogenic transformation, i.e. anchorage-independent growth, loss of

contact inhibition.

B. Increased motility and invasiveness of cancer cell lines

C. Cell transformation

Activation of a telomerase that prevents loss of telomere length, leading

to immortalization of cells.

10. Alters cell proliferation,

cell death or nutrient supply

Interference with cell-signaling pathways leading to expression of

carcinogenic trait/phenotype in the cell e.g. facilitating cell invasion or

induction of gene promotion for inflammatory mediators, oncogenes.

Induced defects in programmed cell death (apoptosis). Evasion of

apoptosis is a requirement for both neoplastic transformation and

sustained growth of cancer cells.

Detection of alterations in cell proliferation and cell-cycle effects (e.g.

DNAreplication changes, cell-cycle control, ploidy), mitogenesis. Altered

nutrient supply affects cell viability.

Change in pro-angiogenesis factors

Disruption of gap-junction intercellular communication pathways that

can cause a loss of ‘contact inhibition’ and abnormal cell growth.

The bystander effect was first identified in radiobiology and refers to the

situation where non-irradiated cells exhibit effects caused by radiation

as a result of chemical signals (messengers) received from nearby

irradiated cells. These effects are often mediated through gap-junction

transfer of chemical agents.

Any of the 10

characteristics in this table

could interact with any

other (e.g., oxidative

stress, DNA damage, and

chronic inflammation),

which when combined

provides stronger evidence

for a cancer mechanism

than would oxidative stress

alone.

Any of the 10 characteristics in this table could interact with any other (e.g., oxidative stress, DNA

damage, and chronic inflammation), which when combined provides stronger evidence for a cancer

mechanism than would oxidative stress alone.

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174

APPENDIX 9: GUIDELINES ON MICROBIOLOGICAL QUALITY OF THE

FINISHED COSMETIC PRODUCT

This part has been taken over from the 9th Revision of the NoG (SCCS/1564/15):

https://ec.europa.eu/health/sites/health/files/scientific_committees/consumer_safety/docs

/sccs_o_190.pdf

Although the NoG are concerned with the safety evaluation of ingredients, this appendix is

concerned with the finished cosmetic product. The reason for this is the fact that in other

pieces of legislation, reference has been made to it as being part of the NoG.

Preamble:

Skin and mucous membranes are protected from microbial attack by a natural mechanical

barrier and various defence mechanisms. However, these may be damaged and slight

trauma may be caused by the action of some cosmetics that may enhance microbial

infection. This may become of particular concern when cosmetics are used around the eyes,

on mucous membranes in general, on damaged skin, on children under 3 years, on elderly

people and persons with compromised immune system. Consequently, two separate

categories of cosmetic products are defined in the microbiological quality control limits:

Category 1: Products specifically intended for children under 3 years, to be used in the eye

area and on mucous membranes.

Category 2: Other products.

Microbial contaminants usually come from two different origins: during production and filling,

and during the use of the cosmetic by the consumer. From the moment the cosmetic unit is

opened until the last use of the product by the consumer(s), a permanent, variable and

additive microbial contamination of the cosmetic is introduced, caused by the domestic

environment and contact with the skin of the consumer(s) (hands and body).

Reasons for microbial preservation of cosmetics are:

- to ensure the microbial safety of cosmetics for the consumer,

- to maintain the quality and specifications intended of the product,

- to confirm hygienic and high-quality handling.

Although only a small number of cases of microbiological contamination of cosmetics leading

to microbial infections of the consumer has been reported, microbial contamination of

cosmetic products may spoil them or seriously reduce the intended quality. In order to

ensure the quality of the product and the safety for the consumer, it is necessary to carry

out routine microbiological analysis of each batch of the finished product coming on the

market. In some justified cases (e.g. alcohol content > 20%), end product testing is not

necessary (ISO 29621, 2010). The parameters examined, the criteria and methods used,

and the results obtained per batch should be specified in properly filed reports and be taken

up in the TIF.

Quantitative and qualitative limits

Quantitative and qualitative limits are based on the European Standard EN ISO 17516:2014

Cosmetics – Microbiology – Microbiological limits. The European Standard EN ISO

17516:2014 was approved by CEN on 9 August 2014 and at present is widely used by the

cosmetics industry as international standard (Table A9). It is reviewed and confirmed in

2020

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175

Table A9: Microbiological limits for cosmetics. European Standard EN ISO 17516:2014

Cosmetics –Microbiology – Microbiological limits

Types of microorganism

Products specifically

intended for children

under three years of age,

the eye area or the

mucous membranes

Other products

Total Aerobic Mesophilic

Microorganisms (Bacteria

plus yeast and mould)

≤ 1 x 10

CFU per g or

≤ 1 x 10

CFU per g or

Escherichia coli

Absence in 1 g or 1 ml

Pseudomonas aeruginosa

Absence in 1 g or 1 ml

Staphyloccocus aureus

Absence in 1 g or 1 ml

Candida albicans

Absence in 1 g or 1 ml

Due to inherent variability of the plate count method, according to USP Chapter 61 or EP

Chapter 2.6.12, Interpretation of results, results considered out of limit if

a > 200 CFU/g or ml,

b > 2 000 CFU/g or ml.

NOTE When colonies of bacteria are detected on Sabouraud Dextrose agar, Sabouraud

Dextrose agar containing antibiotics may be used.

Challenge testing (based on US Pharmacopoeia 2014, European Pharmacopoeia 2014)

Note that this chapter addresses microbiological contamination, i.e. unwanted presence of

microorganisms. Total germ counts and challenge test are not directly applicable for the case

of probiotic cosmetic formulations to which live or viable microorganisms have been

deliberately added.

The efficacy of the preservation of a cosmetic product under development has to be assessed

experimentally in order to ensure microbial stability and preservation during storage and

use. This is done by challenge testing. The latter is mandatory for all cosmetic products that,

under normal conditions of storage and use, may deteriorate or form a risk to infect the

consumer.

A challenge test consists of an artificial contamination of the finished product, followed by a

subsequent evaluation of the decrease in contamination to levels ensuring the microbial

limits established for Categories 1 and 2. The microorganisms used in the challenge test may

be issued from official collection strains from any state in the EU to ensure reproducibility of

the test and are: Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans and

Aspergillus brasiliensis.

It is well known today that the consistency of challenge tests relies more on the capability

of the used microorganisms to contaminate a specific cosmetic product than on the

taxonomic status of the microorganisms, their initial concentrations, or the conditions of

incubation and media of recovery used. Microorganisms with the capability to contaminate

specific cosmetics are the best candidates for use in a challenge test. The microbicidal

activity of preservatives or any other compound in the finished cosmetic must be ruled out

in the challenge test by dilution, filtration, addition of neutralisers or any other means.

The experimental performance of the microbial controls and the challenge tests must be

carried out/supervised and validated by a microbiologist. As mentioned before, the

responsible person must guarantee the efficacy of the preservation of his products

experimentally by challenge testing. However, as no legal or universal challenge test method

is currently available, it is up to the responsible person to decide on the details of the test

to be used.

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176

Good Manufacturing Practice (GMP)

In order to comply (mandatory but no certification required) with Good Manufacturing

Practice and Microbial Quality Management, manufacturers of cosmetics have to define and

follow specific cleaning, sanitation and control procedures to keep all apparatus and materials

appropriately clean and free of pathologic microorganisms. Procedures also include

microbiological control of raw materials, bulk and finished products, packaging material,

personnel, equipment and preparation and storage rooms. Compliance should be checked

with the currently available European Committee for standardization (CEN) standards

(available through http://www.cenorm.be/cenorm/index.htm) and/or ISO standards

(available through http://www.iso.org/iso/en/ISOOnline.frontpage). According to Article 8 of

Regulation (EC) No 1223/2009, good manufacturing shall be presumed where the

manufacture is in accordance with the relevant harmonised standards, the references of

which have been published in the Official Journal of the European Union.

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177

APPENDIX 10: FREE ACCESS TO IN SILICO MUTAGENICITY /

GENOTOXICITY AND CARCINOGENICITY DATABASES

 The Danish QSAR database (http://qsar.food.dtu.dk/) which includes QSAR models

based on structural alerts for DNA reactivity; in vitro Ames test in S. typhimurium,

chromosome aberration in Chinese Hamster Lung (CHL) and ovary (CHO) cells;

Comet assay in mouse; micronucleus test in mouse erythrocytes; sister chromatid

exchange in mouse bone marrow cells; mutations in HGPRT locus in Chinese hamster

ovary (CHO) cells; mutations in thymidine kinase locus in mouse lymphoma cells;

and sex-linked recessive lethal test in Drosophila melanogaster.

 The OECD QSAR Toolbox (https://qsartoolbox.org/), which also incorporates the

models and tools from the Danish QSAR database, provides a versatile suite of

programs for chemical profiling, categorisation, and data gap filling by (Q)SAR models

and read-across for various toxicological endpoints, including mutagenicity. The

system also includes metabolic simulators that further enable the prediction and

genotoxicity assessment of metabolites. The Toolbox also provides profilers for

mutagenicity that are based on structural alerts for in vitro mutagenicity (Ames test),

in vivo mutagenicity (micronucleus) chromosomal aberration and micronucleus test,

and DNA and protein binding. The predictions from the profilers can provide

supporting information when used in conjunction with QSAR predictions. The Toolbox

also provides a few profilers that combine several structural alerts for the purpose of

category formation on the basis of carcinogenicity potential of chemical substances.

A notable one is the ISS profiler that combines 58 structural alerts for carcinogenicity

(both genotox and non-genotox) from the Toxtree software

(http://toxtree.sourceforge.net/ ). Around 20 of the alerts are for non-genotoxic

carcinogenicity, and the remaining ones for genotoxic carcinogenicity (mutagenicity).

A recent study (Aljallal, 2020) has indicated that some of the structural alerts and

the profilers provided in the OECD QSAR Toolbox need further refinement, and their

use in conjunction with QSAR models and read-across would be required to improve

the accuracy of predictions.

 VEGA QSAR platform (www.vegahub.eu/) provides QSAR models for mutagenicity

developed in line with the OECD principles using high quality datasets with the aim

to use for regulatory purposes;

 The US-EPA’s Toxicity Estimation Software Tool (T.E.S.T.)

(www.epa.gov/nrmrl/std/qsar/qsar.html) is an Expert system that uses an ensemble

of QSARs to estimate toxicity - including mutagenicity (Ames test in S. typhimurium);

 Toxtree (http://toxtree.sourceforge.net/) enables estimation of toxicity hazard by

applying a decision tree approach;

 Lazar (https://lazar.in-silico.ch/predict) is an automated system of read across to

calculate toxicity predictions.

 OpenTox for carcinogenicity through OpenTox platform (ToxPredict)

(www.opentox.net/library/toxicity-prediction

 OncoLogic (US EPA) (www.epa.gov/tsca-screening-tools/oncologictm-expert-

system-evaluate-carcinogenic-potential-chemicals

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178

APPENDIX 11: INHALATION PARAMETERISATION

Table A. 11: Example for the parameterisation of a 2- Box model for sunscreens based on

Rothe et al., 2011 and SCCS recommendations. Product-dependent parameter values in this

example are specific for sunscreens and denoted with an asterix * (see also 3-3.5.4.1

calculation of the inhalation SED)

Parameter

Parameter description

Propellant

spray

Pump

spray

Unit

Reference

product

amount per application*

g/application

SCCS, NoG

air

air-borne fraction of spray

mist

0.2

fraction

Bremmer et al, 2006

Box 1 (Near-field around

the head)*

1000

SCCS, Octocrylene

duration of exposure in

Box 1

(near field)*

min

SCCS, Octocrylene

inh

inhalation rate

13**

L/min

US-EPA 2011

Box 2 (Far-field, e.g.

bathroom)*

10000

SCCS, Octocrylene

duration of exposure in

Box 2 (far field)*

min

SCCS, Octocrylene

resp

respirable fraction

experimental

data

experimental

data

fraction

ret

substance retention

fraction in the lungs (25%

exhaled)

0.75

fraction

Rothe et al., 2011

appl

frequency of application*

per day

SCCS, NoG

bodyweight

SCCS, NoG

* Product-dependent parameter value;

**highest median among several adult age categories;

SCC/1627/21 opinion on octocrylene; SCCS NoG = SCCS Notes of Guidance.

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179

APPENDIX 12: LIFETIME CANCER RISK (LCR) APPROACH

The "T25 method" (Sanner et al., 2001) is used as a simple method for quantitative risk

assessment of carcinogens in the REACH Regulation (ECHA, 2017). It should be noted that,

in six cases where high quality epidemiology and animal carcinogenicity studies were

available, quantitative risk characterisation based on epidemiological data and data based on

animal studies using the T25 method differed by factors of less than three (Sanner and

Dybing, 2005).

Determination of the LCR is carried out in different steps. After having decided what animal

data set to use and type of tumour to consider, the dose descriptor T25 is determined, which

is described in detail (ECHA, 2012a; Dybing et al., 1997).

The animal dose descriptor (T25) is converted to the human dose descriptor (HT25) based

on comparative metabolic rates (Sanner et al., 2001):

HT25=

T25

(body weight

human

/body weight

animal

)

0.25

Based on the daily lifetime SED, the LCR is calculated by linear extrapolation by use of the

following formula:

LCR =

SED

HT25/0.25

Subsequently, a statement is generated describing whether the actual risk may be higher or

lower than the risk calculated for a specific scenario. The procedure and the following

elements are reported and discussed in detail (Sanner et al., 2001; ECHA, 2012a).

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180

APPENDIX 13: POD USED FOR TTC DERIVATION

Table A.13: Chemical classes and 5

percentile Cramer Class PoDs of selected published TTC

datasets. n = number of substances / PoDs in the dataset. 5

percentiles were derived from log-normal

parametric distributions, except by Pinalli et al., (2011); van Ravenzwaay (2017) and Kalkhof et al.

(2012) who did not report the calculation method. NO(A)EL = No Observed Adverse Effect Level; PoD

= Point of Departure; CC = Cramer Class; DB = data base; dev. = developmental

References

Dominant chemical classes

5th percentile NO(A)ELs

adjusted to chronic (PoD,

mg/kg bw/day)

Cramer

Class I

Cramer

Class II

Cramer

Class III

Yang et al., 2017,

‘federated’

Cosmetic-related,

packaging and pesticides

977

4.57

0.62

0.23

Munro et al., 1996

Food contact, pesticides

612

3.0

0.91

0.15

Yang et al., 2017, Munro

et al., 1996 ‘Munro-2016’

Some Cramer Classes and

other errors corrected,

harmonised assessment

factors

612

4.90

1.07

0.15

Yang et al., 2017,

COSMOS 2017

Cosmetic-related & packaging

552

4.20

0.58

0.79

Tluczkiewicz et al., 2011

Industrial chemicals and

pesticides

521

0.62

0.038

Kalkhof et al., 2012

German pre-REACH DB,

industrial chemicals

813

2.5 (n=69)

2.5 (n=20)

1.3

(n=724)

Pinalli et al., 2011

Food contact materials

232

CCI/II not reported

CCIII reported in

Feigenbaum et al.,

2015

0.4

(n=113)

Feigenbaum et al., 2015

Pesticides without carbamates

and organophosphates

279

0.2

Munro+ Pinalli+ pesticides

with carbamates and

organophosphates

840

0.15

Laufersweiler et al., 2012

Only reproductive and

developmental endpoints.

From Kroes et al. (2000),

Bernauer et al. (2008), plus

literature.

283

13.1

(n=69)

1.87

(n=11)

0.31

(n=203)

Van Ravenzwaay et al.,

2017

Chemicals & pesticides,

developmental studies only

150§/

537*

§rabbits: 5/9.5 maternal/dev

*rats: 7.6/10 maternal/dev

Patel et al., 2020

‘RIFM’

Fragrance chemicals

476

5.39

(n=238)

1.97

(n=76)

1.17

(n=162)

Patel et al., 2020

‘RIFM/COSMOS/Federated’

Fragrance chemicals,

Cosmetic-related, packaging

and pesticides

1327

4.91

(n=421)

1.27

(n=111)

0.29

(n=795)

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181

ABBREVIATIONS AND GLOSSARY OF TERMS

2D Two Dimensional

Three Dimensional

Refinement, Reduction, Replacement

3T3 NRU PT

3T3 Neutral Red Uptake Phototoxicity Test

Androgen

Angström

ADME

Absorption, Distribution, Metabolism, Excretion

ADRA

Amino acid Derivative Reactivity Assay

Adverse

An adverse response is defined as any treatment-related

response that results in change in the morphology,

physiology, growth, development or life-span of an organism,

which results in an impairment of functional capacity, an

impairment of the capacity to compensate for additional

stress, or an increase in susceptibility to other environmental

influences (WHO 2004)

AEL

Acceptable Exposure Level

AhR

Aryl hydrocarbon Receptor

AIC

Akaike Information Criterion

A.I.S.E.

International Association for Soaps, Detergents and

Maintenance Products

Alternative methods

All those procedure which can completely replace the need

for animal experiments, which can reduce the number of

animals required, or which can reduce the amount of pain

and stress to which the animal is subjected in order to meet

the essential needs of humans and other animals

(Rogiers et al., 2000; Russell et al., 1959)

AMA

Amphibian Metamorphosis Assay

AOP

Adverse Outcome Pathway

Androgen Receptor

Art.

Article

AhR

Aryl hydrocarbon Receptor

ATM

Alternative Test Method

ATP

Adaptation to Technical and scientific Progress

AUC

Area Under the Curve

BCOP

Bovine Corneal Opacity and Permeability

BCRP

Breast Cancer Resistance Protein

BHT

Butylated HydroxyToluene

BMD

The BenchMark Dose is proposed as an alternative for the

classical NOAEL and LOAEL values. The BMD is based on a

mathematical model being fitted to the experimental data

within the observable range and estimates the dose that

causes a low but measurable response (the benchmark

response BMR) typically chosen at a 5 or 10% incidence

above the control.

BMDS

BMD Software

BMDL

BMD Lower limit refers to the corresponding lower limit of a

one-sided 95% confidence interval on the BMD.

BMDU

BMD Upper limit refers to the corresponding upper limit of a

one-sided 95% confidence interval on the BMD.

BMR

BenchMark Response

BoA

Board of Appeal

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182

BrdU

5-Bromo-2-deoxy-Uridine

BSE

Bovine Spongiform Encephalopathy

Body Weight

Concentration

CAS n°

Chemical Abstracts Service registry number

Cat.