FRAME Response to the European CommissionConsultation on Cosmetics Testing Methods
A FRAME Response to the European Commission Consultation on the Draft Report on Alternative (Nonanimal) Methods for Cosmetics Testing: Current Status and Future Prospects
ATLA 38, 345�353, 2010 345 Comment A FRAME Response to the European Commission Consultation on the Draft Report on Alternative (Nonanimal) Methods for Cosmetics Testing: Current Status and Future Prospects -- 2010a Michael Balls and Richard Clothier Fund for the Replacement of Animals in Medical Experiments (FRAME), Nottingham, UK Summary -- This response on behalf of FRAME to the European Commission's consultation on the five chapters of the Draft Report on Alternative (Non-animal) Methods for Cosmetics Testing: Current Status and Future Prospects -- 2010, is via a Comment in ATLA, rather than via the template supplied by the Commission. This is principally so that a number of general points about cosmetic ingredient testing can be made. It is concluded that the five draft chapters do not provide a credible basis for the Commission's forthcoming report to the European Parliament and the European Council on the five cosmetic ingredient safety issues for which the 7th Amendment to the Cosmetic Directive's ban on animal testing was postponed until 2013. This is mainly because there is insufficient focus in the draft chapters on the specific nature of cosmetic ingredients, their uses, their local effects and metabolism at their sites of application, and, in particular, on whether their possible absorption into the body would be likely to lead to their accumulation in target sites at levels approaching Thresholds of Toxicological Concern. Meanwhile, there continues to be uncertainty about how the provisions of the Cosmetics Directive should be applied, given the requirements of the REACH system and directives concerned with the safety of other chemicals and products. Key words: alternative methods, animal tests, carcinogenicity, cosmetic ingredients, Cosmetics Directive, in silico, in vitro, REACH system, repeated dose testing, reproductive toxicity, skin sensitisation, toxicokinetics. Address for correspondence: Michael Balls, FRAME, Russell & Burch House, 96�98 North Sherwood Street, Nottingham NG1 4EE, UK. E-mail: email@example.com Introduction In July 2010, the European Commission indicated a desire to consult the public on the five chapters of a Draft Report on Alternative (Non-animal) Methods for Cosmetics Testing: Current Status and Future Prospects -- 2010, in order "to ensure that each chapter correctly reflects the current state of the art and the prospects". This draft report was prepared by working groups of experts nominated by the various stakeholders and chaired by the Commission's Joint Research Centre (JRC), in order to gain a broad and objective picture of the scientific/technical issues that relate to establishing alternative test methods for the five human health(-related) effects falling under the 2013 deadline for the marketing ban of the EU Cosmetics Directive. It was also intended that the draft report would contain, where possible, a sciencebased estimate of the time necessary to achieve full replacement of animal testing for the respective endpoints. Directive 76/768/EEC (the Cosmetics Directive) is a regulatory framework for the placing on the market of cosmetic products, and subsequent amendments and associated directives have aimed to phase out the use of animal testing for these purposes. Besides the complete testing ban, a marketing ban has applied since 11 March 2009, for all human health(-related) effects, with the exception aThe authors gratefully acknowledge their very useful discussions with Professor Horst Spielmann (Department of Biology, Chemistry and Pharmacy, Free University of Berlin, Berlin, Germany) during the preparation of this Comment. 346 Comment of five issues, namely, repeated dose toxicity (including skin sensitisation and carcinogenicity), reproductive toxicity and toxicokinetics, for which the deadline was extended to 11 March 2013. The consultation is related to a requirement that the Commission must inform the European Parliament and the European Council "in case alternative methods will not have been developed and validated by 2013 for the remaining endpoints that are exempted from the Cosmetic Directive's marketing ban until 2013". The introductory documents state that "The public is invited to comment on the five individual chapters of the draft report, each representing one human health(-related) effect. The focus of the consultation is to ensure that each chapter correctly reflects the current state of the art and the prospects. As regards prospects, the experts evaluated how long it will take to develop and optimise such approaches up to a level that fulfils the ECVAM criteria for entering prevalidation. The time required for the validation and regulatory acceptance of these alternative approaches was not considered for the purpose of this exercise. In addition, justified comments on the conclusion (i.e. the timelines given to achieve full replacement) are sought." It is clearly indicated that "The Commission is inviting factual comments, which complement the information provided. Comments should include references and other substantiation where possible." "Any comments and information on this public consultation should be submitted by using the provided comment template form by mail, fax or e-mail, by 15 October 2010 at the latest, to: European Commission, Health and Consumers Directorate-General (DG SANCO), Unit SANCO B2, Cosmetics and Medical Devices, B-1049 Brussels, Belgium (Fax: 00 32 (0) 2 296 64 67; E-mail: SANCO-COSMETICS-REPORT@ec.europa.eu)". The template requires the submitters of comments to indicate their names, affiliations and types of organisation, the submission date, and whether the submission should/should not be treated as confidential. A separate template form should be submitted for each of the five chapters, and the forms have four columns, for "comment number", "line number", "comment and rationale", and "proposed revised text". The introductory documents, the five draft chapters, and the template are available at: http://ec. europa.eu/consumers/sectors/cosmetics/documents/ public_consultation/index_en.htm FRAME's Submission We have decided not to respond to the consultation by using the template system, but to convey our views via a Comment in ATLA, principally because there are a number of important general points to be made, which could not readily be made via the "line number/comment and rationale/proposed revised text" template system. In addition, a detailed set of informed comments would inevitably involve many pages for each draft chapter, and it is not clear how the Commission or the authors of the chapters would handle large numbers of suggestions for "proposed revised texts". We also doubt whether more than a handful of members of "the public" would have the competence or the information necessary for providing assurance that a chapter "correctly reflects the current state of the art and the prospects". People more cynical than ourselves would undoubtedly be tempted to consider that the consultation has deliberately been set up in this way, in order to discourage participation. In our analysis of the draft chapters, we will focus on what the Commission will need, if it is to provide the European Parliament and the European Council with information of the necessary quality, which is specifically relevant to the safety of cosmetic products, their ingredients, their purposes and the consequences of repeated exposure to them, in relation to the Cosmetics Directive, rather than on issues related to chemicals in general and to other products, which are regulated by the REACH system and other directives. In particular, it should be remembered that a cosmetic product is defined as "any substance or preparation intended to be placed in contact with the various 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 and/or correcting body odours and/or protecting them or keeping them in good condition" (1). By contrast, a medicinal product is defined as "a) any substance or combination of substances presented as having properties for treating or preventing disease in human beings; or b) any substance or combination of substances which may be used in or administered to human beings, either with a view to restoring, correcting or modifying physiological functions by exerting a pharmacological, immunological or metabolic action, or to making a medical diagnosis" (1). For the purposes of this Comment, we will go along with the current practice of the Commission and the European cosmetics industry, by avoiding the term `cosmeceutical', which refers to cosmetic products containing biologically active ingredients which are claimed to have medical or drug-like benefits. Nevertheless, this issue will have to be faced up to at some time in the future, since, if the claims increasingly being made about them are genuine, these products would have to be treated as medicinal products rather than cosmetics, in the Comment 347 interests of avoiding damage to human health. A crucial question concerning cosmetic ingredients, is the extent to which they are absorbed across the skin or other barriers which limit the entry of xenobiotics into the body (e.g. barriers in the lungs, buccal cavity and vagina, which, in this case, are more important than the gastric or intestinal barriers). Metabolism is often referred to (especially in pointing out the limitations of in vitro test systems), but it is metabolism in the skin and the other relevant barriers which may be of particular concern, since the importance of metabolism by visceral organs, such as the liver and kidneys, will depend on the site of exposure, the route of absorption and transport to them. Surely, it is reasonable to assume that cosmetic products are designed so that significant amounts of their ingredients are not taken into the body, so as to exceed Thresholds of Toxicological Concern (TTCs) and adversely affect the visceral organs or systems. Another important criterion in evaluating the draft chapters is the way in which they deal with the limitations of animal models and of the current Organisation for Economic Co-operation and Development (OECD) Health Effects Test Guidelines (TGs), the relevance of which tends to be compromised by species differences and the application of very high doses. How the data they provide can be interpreted rationally in terms of potential hazard, exposure and risk, and actual experience, in humans, not forgetting the high levels of qualitative and quantitative variation in responses to chemicals in the human population, is a vital component of the background against which non-animal methods and testing strategies for evaluating the safety of cosmetic ingredients should be developed, validated and applied. Draft Chapter 1: Repeated Dose Toxicity This 42-page draft chapter, by Stuart Creton, Pilar Prieto, Alan Boobis, Wolfgang Dekant, Jos Kleinjans, Hannu Komulainen, Paolo Mazzatorta, Anna Price, Vera Rogiers, Greet Schoeters and Mathieu Vinken, focuses on the main available in vitro models in relation to six of the most common targets for toxicity (liver, kidney, central nervous system, lung, cardiovascular system, and haematopoietic system). It is concluded that complete replacement will not be possible by 2013. Various OECD TGs for repeated dose toxicity are discussed, and it is noted on line 136 that "the 90-day oral toxicity in rats is the most commonly conducted repeated dose toxicity study" for cosmetic ingredients, which is mostly used to derive the No Observable Adverse Effect Level (NOAEL) for cosmetic ingredients. The NOAEL is used in the calculation of the MoS (Margin of Safety) and/or the Margin of Exposure (MoE) for cosmetic ingredients (line 91). There is no mention of OECD TGs 427 (2) and 428 (3), on skin absorption in vivo and in vitro, respectively, and the assumption seems to be that the relevant dose is that which comes within the compass of the animal tests, rather than the amount of a compound which is likely to reach these six most common targets as a result of repeated exposure to the cosmetic product. Surely, it would be more useful to think in terms of a Margin of Uptake (MoU), i.e. the difference between the likely uptake of a cosmetic ingredient and the level in the body which would be a cause for concern (i.e. the TTC). The skin seems to be regarded merely as a route of entry into the body in general, as there is no mention of dermal toxicity per se -- none of the six tables on alternative methods for repeated dose toxicity deal with the skin. It is also interesting to note (line 598) that the liver is the main focus of the current COLIPA co-funded projects on non-animal methods for assessing repeated dose toxicity. Nevertheless, much is being achieved in other toxicological studies with reconstituted human skin equivalents, which might well prove to be suitable for use in repeated dose studies. Little attention is paid to the kinds of chemicals which are used as cosmetic ingredients, and which, on the whole, unlike pharmaceuticals, are not intended to be readily taken up into the bloodstream for distribution to internal organs. Indeed, the authors of this draft chapter would appear to be more comfortable in dealing with issues related to the use and testing of pharmaceuticals. This is particularly true of the section on the concept of Mode of Action (MoA) in the induction of adverse health effects, which might be better applied in its development stage to compounds likely to be highly toxic, rather than to cosmetic ingredients, which would overwhelmingly be expected to be toxicologically inert at the levels to which users of cosmetics are exposed. There is little information on how the cosmetic industry deals with the issues raised by repeated exposure to their products, and it is regrettable (line 677) that the Working Group received little response to their enquiry via COLIPA as to whether companies would share their own experiences with illustrative examples. The paragraph outlining the Unilever approach is encouraging (especially when considered with the 2009 review by Carmichael et al. , which is not listed in this draft report), but the short Nestl� section, although it does, at least, mention how exposure is taken into account, is more relevant to food products than to cosmetics. Other companies, such as Beiersdorf, L'Or�al, and Procter and Gamble, have invested huge sums in the development of non-animal methods -- it would have been good to see 348 Comment some input from them as well. In particular, it would be interesting to ask the cosmetic industry for information known to them concerning cosmetic ingredients which are taken into the body at levels which could result in damage to the six targets which are the focus of this draft chapter. It should be noted, for example, that, if damage to the nervous system were to be caused, such damage would be more likely to affect the peripheral nervous system than the central nervous system. There are in vitro methods for looking into this kind of problem (e.g. 5, 6). The draft chapter mentions some of the limitations of animal models, but rather than using the structure of the placenta as an example of an anatomical difference between animal and humans (line 763), it would have been more relevant to cosmetic ingredient toxicity/safety to draw attention to differences between animal and humans in the structure of the skin, eyes and lungs. Some regret is expressed that larger numbers of animals, sufficient to detect rare events, cannot be used, but it would, in any case, be very difficult to assess the relevance of such rare cases to the human situation. The same could be said of human studies, but there is undoubtedly a case for making greater use of human experience, including ethically-conducted human trials, and of post-marketing surveillance (other than for marketing purposes). We will make rather few comments on the sections on non-animal methods, since their use could only be considered meaningfully against a sufficient understanding of cosmetics and their ingredients. There are a few references to in silico systems in the draft chapter, and the focus is on the six common targets and the Maximum Recommended Therapeutic Dose (MRTD), which will tell the Commission very little of relevance to cosmetic ingredients. This is a lost opportunity, since in silico methods could be expected to have great value in the future, as ways of predicting whether cosmetic ingredients would be likely to be metabolised and/or have adverse effects at their sites of application, and/or be absorbed into the body in significant amounts relevant to TTCs. It is concluded in the Executive Summary (line 59) that none of the available in vitro methods are suitable for quantitative risk assessment for repeated dose toxicity. Since much of the focus of the draft chapter is on approaches or systems in the course of development, that is an oversimplification. In fact, methods have been developed which permit repeated assays to be performed on the same cell cultures, and this opens the possibility of studies on the extent of recovery from adverse effects (7, 8). Squamous differentiation in bronchial epithelial cell cultures can be studied by using a non-cytotoxic endpoint involving fluores- cent cadaverine (9, 10). The possibility of repeated treatment of the same cell cultures has also been exploited in studies on the repeated effects of radiation (11). These methods permit studies on recovery from damage, and on the effects of cosmetic ingredients on recovery. Overall, then, whatever its value in relation to repeated dosage and visceral target organ toxicity, this draft chapter will provide the Commission with little information of value concerning the application of the ban on the animal testing of cosmetic ingredients. Draft Chapter 2: Skin Sensitisation The conclusion in this 24-page draft chapter, by David Basketter, Silvia Casati, Klaus Ejner Andersen, Alexandre Angers-Loustau, Aynur Aptula, Ian Kimber, Reinhard Kreiling, Henk van Loveren, Gavin Maxwell and Hanna T�hti, is that no complete replacement of the in vivo methods will be available by 2013, but that it should be possible by 2017�19 "to make skin sensitisation risk assessment decisions by using a toolbox of non-animal test methods for all cosmetic ingredients and exposure scenarios". Understandably, perhaps, since its area of coverage is more-tightly defined, this is a much more useful draft chapter. It demonstrates that very good effort is being focused on increasing our understanding of the induction and expression of skin allergies, and indicates how this new knowledge can be applied to predicting the skin sensitiser potency of cosmetic ingredients. There is a good balance between the consideration of in silico tools and in vitro assays in the assessment of key steps in the initiation of an allergic response. The in silico approaches could be particularly valuable in predicting various effects of the structures of chemicals, including haptenation (the covalent binding of a chemical to a skin protein), upon which sensitisation potential depends, and metabolism in the skin, as well as skin sensitising potential itself. Knowing that a chemical would not bind to protein and that it could not be metabolised by enzymes in the skin, would be of great value. Much is made of the value of the local lymph node assay (LLNA) in predicting the results of the human repeated insult patch test (HRIPT), which is somewhat surprising, since the evidence from other toxicological studies is that species variation between animal models and humans limits the value of animal data for predicting human effects. That could be because the good correlations have mainly involved the most highly-potent sensitisers, where, perhaps, the power of the chemical in the reaction overwhelms the variability in the Comment 349 responding system which may be involved in the wide range of sensitivities to allergens in the human population. It is extremely unlikely that such highly-potent sensitisers, which would affect the human population in general, would be used as cosmetic ingredients. The use of in vitro systems has great promise, as is reflected in this draft report. One recent study is worth mentioning here, which involved a comparison of the human cell-line activation test (h-CLAT) and the LLNA, in which 100 chemicals were tested independently by two laboratories (12). The test set included chemicals classified as extreme, strong, moderate and weak sensitisers, and nonsensitisers, in the LLNA. The correlation between the results of the two tests was 84%, and reasons for the difference were discussed. The draft chapter would benefit from an indication of how the various approaches in use, or in the course of development, are, or could be, used in a stepwise, decision-tree strategy for making decisions about the sensitising potential/safety of cosmetic ingredients. Also, the overall conclusion about the time needed to achieve the replacement of animal tests may be unduly pessimistic. It is to be hoped that this is not because there is satisfaction that the reduction and refinement aspects of the LLNA are sufficient to justify its continued use in the long term. Draft Chapter 3: Carcinogenicity This 38-page draft chapter, by Jan van Benthem, Susan Felter, Tuula Heinonen, Albrecht Poth, Rositsa Serafimova, Joost van Delft, Emilio Benfenati, Pascal Phrakonkham, Andrew Worth and Raffaella Corvi, is based on the belief that "the twoyear bioassay in rodents is widely regarded as the gold-standard to evaluate cancer hazard and potency", and regrets that "this test is rarely done on cosmetic ingredients because of expense, time, and animal welfare issues" (lines 43 and 91). Shortterm in vivo and in vitro tests for genotoxicity are seen as inadequate substitutes, along with a 90-day repeated dose test for non-genotoxic carcinogens (line 94). In the Executive Summary (line 67), it is concluded that "Taking into consideration the present state of the art of the non-animal methods, the experts were unable to suggest a timeline for full replacement of animal tests currently needed to fully evaluate carcinogenic risks of chemicals. Although a timeline for full replacement cannot be developed, clearly the timeline is expected to extend past 2013." It is further concluded that "animal testing bans under the 7th Amendment to the Cosmetic Directive will have a profound effect on the ability to evaluate and conduct a risk assessment for potential carcinogenicity of new cosmetic ingredients" (lines 49, 97 and 1103). It is amazing that such opinions could be recorded in 2010, in what should be an authoritative document, and this draft chapter could not possibly be a reliable basis for the advice to be given by services of the Commission to the Parliament and the Council. It is now widely felt that animal carcinogenicity data are not adequate to support the classification of a chemical as "probable human carcinogen" or "probable non-carcinogen" (13, 14). Indeed, the data from the bioassay in one rodent species (e.g. the rat) cannot be used to predict what would be expected to happen in another rodent species (e.g. the mouse), so the lack of applicability of rodent data to humans is not surprising. This is because, in addition to significant physiological differences between humans and laboratory animals, the lifetime doses applied in the rodent bioassay are very high, and are indeed much higher than could possibly be experienced in humans, particularly in the case of cosmetic ingredients. There is a danger that some toxicologists would settle for in silico and in vitro approaches which could provide the identifications and classifications currently provided by the rodent bioassay. Indeed, the in vitro cell transformation assays, some of which have been in the course of development and evaluation for decades, are based on rodent cell cultures. What is needed is a complete rethink, with the aim of developing methods for identifying potential human carcinogens. Hopefully, these methods could employ human material and/or human experience, and could be based on understanding of key events in the carcinogenic process. Meanwhile, particular attention should be focused on the likelihood that some cosmetic ingredients could be carcinogenic in humans, but this would require the paying of rigorous attention to the nature of the chemicals themselves, their effects at their sites of exposure, their uptake into the body, and the levels at which they might be transported to internal body tissues. The TTC concept is very important here, and it is well summarised in the draft chapter (line 522). Insufficient attention is paid to the skin as a target organ for genotoxins and carcinogens, though the use of human reconstructed epidermis in vitro models for genotoxicity testing in, for example, assays such as the micronucleus assay and the Comet assay, is discussed (lines 701 and 731). The need to focus on cosmetic ingredients themselves should be clear. Most of them will be innocuous in terms of all kinds of toxicity, but some of them will be potent chemicals which will need close attention. This latter group includes the colourants, including hair dyes, UV filters (sunscreens) and preservatives, which are specifically referred to in the Cosmetics Directive. It was 350 Comment the positive results obtained with hair dye components in bacterial tests for mutagenicity (e.g. the Ames Test), which led to the emphasis on genotoxicity testing in the application of the Directive in relation to cosmetic products ingredients, and to the inclusion of so many genotoxicologists among the experts who were directly involved or consulted. Little attention is paid to cosmetic ingredients per se in the draft chapter. There is a brief discussion on hair dyes, but only to make the point that the in vitro systems used in a study failed to predict the results of the rodent bioassay, so the `falsepositive' results they provided might have led to the unnecessary abandonment of the use of 17 hair dyes. However, given the poor record of predictions based on the rodent bioassay, the in vitro results might have been `true-positive' for humans. As in the case of draft Chapter 1, this draft chapter focuses too much attention on the tests themselves, rather than on the chemicals that need to be considered, how the body is exposed to them, and where and in what circumstances they might reach a TTC. As a result, the statement that, if "an in vivo test is no longer possible, the safety of many potential new cosmetic ingredients will not be able to be substantiated" (line 1103) has no merit and should not influence the application of the 7th Amendment to the Cosmetics Directive. If in vivo testing "is rarely done on cosmetic ingredients because of expense, time, and animal welfare issues" (lines 44 and 92), what problems have arisen as a consequence of this, which now make this a matter of such concern? Draft Chapter 4: Toxicokinetics The 73-page draft chapter, by Olavi Pelkonen, Sandra Coecke, Sofia Batista Leite, Ulrike Bernauer, Jos Bessems, Esther Brandon, Frederic Bois, Ursula Gundert-Remy, George Loizou, Emanuela Testai and Jos�-Manuel Zald�var, is very interesting and encouraging, but it is so heavily based on experience with pharmaceuticals that its relevance to the Cosmetics Directive and 2013 is questionable. For example, Table 1 (five pages) on replacement methods for absorption and bioavailability repeatedly states "developed for pharmaceuticals", "not used for cosmetics" and "suitability for cosmetics should be evaluated". Then Table 2 and the respective text sections (lines 685 to 857) on replacement methods for distribution are mainly focused on partition between blood and tissues, the blood�brain barrier, the blood�placental barrier and the blood�testis barrier -- important issues for pharmaceuticals, but not of primary concern for cosmetic ingredients unless they enter the body in significant amounts which approach the TTC. Figure 2 provides a valuable insight into future human risk assessment with no animal assays, but again, it is likely to be more interesting to the pharmaceutical industry than to the cosmetics industry. The same could be said about the sections on plasma protein binding (lines 634 and 658). The sections on models for bioaccessibility (line 505), absorption (line 534) and bioavailability (line 575) are more relevant, and OECD TG 428 (3), on in vitro methods for determining dermal absorption across animal and human skin, is discussed, but the limitations of these methods are emphasised, as they may not provide suitable input for more-conventional physiologicallybased pharmacokinetic (PBPK) models (which, one might say, are more closely-related to oral uptake). Table 3 and the respective text sections (lines 860 to 1099) on metabolism are mainly concerned with the metabolism of pharmaceuticals by the cytochrome p450 system. The situation is best summed up by words from the draft chapter itself (line 1227): "Since only in cases where a cosmetic ingredient is bioavailable following dermal, oral, or inhalation exposure, [will] further tests on systemic and not just local toxicity be necessary, additional efforts are needed to provide reliable alternative methods to assess the bioavailability after oral and inhalation exposure. Several efforts have been undertaken to improve the reliability [of] alternative test methods available [for] assessing absorption via [the] gut and lungs, but still more work would be required." Surely, however, the key potential route of entry for most cosmetic ingredients is via the skin, and this should have been given more attention in this draft chapter. This criticism is partly countered by the final, short section on "strategic considerations of risk assessment of cosmetic ingredients" (line 1421), and the decision-trees for absorption-based testing (Figure 6) and for (internal) exposure-based testing (Figure 7), which contrast well, at least in terms of relevance, with the schematic representation of a physiologically-based toxicokinetic (PBTK) model for a woman (Figure 3). However, that the authors of the draft chapter are more comfortable with products other than cosmetics re-emerges in the Recommendations section (line 1491), where the need for better models of absorption via the lung and excretion via the kidney is emphasised, and the efficiency of the conventional validation procedure is questioned, and an "expert consensus procedure" is recommended to replace it. However, if these five draft chapters provided by experts for the Commission are anything to go by, that would not be a wise way forward. The validation process need not, and should not, be conventional, but should be scientifically rigorous, adaptable, transparent, and free of bias in support of vested interests. Comment 351 Draft Chapter 5: Reproductive Toxicity In this 45-page draft chapter, by Sarah Adler, Thomas Broschard, Susanne Bremer, Mark Cronin, George Daston, Elise Grignard, Aldert Piersma, Guillermo Repetto and Michael Schwarz, relatively little attention is paid to specific issues related to cosmetic ingredients or products. What is provided is a general review which is perhaps of greater relevance to other chemicals and other products. This is more or less admitted in the Executive Summary (line 59). However, even as a general review, this draft chapter is inadequate. We will return to that point, after making some comments on some other aspects of the draft chapter. As is pointed out (line 127), for substances submitted for inclusion in the positive lists of the Cosmetics Directive, a comprehensive dossier must be provided for evaluation by the Scientific Committee on Consumer Safety. Moreover, When considerable oral intake is expected, or when dermal penetration data suggest a significant systemic absorption, information on toxicokinetics, carcinogenicity and reproductive toxicity "may become necessary" (line 132). However, there appears to be little guidance on what studies may need to be conducted, on how oral uptake and dermal absorption should be estimated for cosmetic ingredients, or on what "considerable" or "significant" levels would make testing "necessary". Nevertheless, it is pointed out that a rat developmental toxicity study according to OECD TG 414 (15) is usually considered sufficient, although data from a one-generation (TG 415) or a two-generation (TG 416) study may be included in the manufacturer's dossier (line 136). An extensive inventory of current animal tests is provided (lines 150 to 414), including the notorious Uterotrophic Bioassay (line 325) and the Hershberger Bioassay (line 356). It should be noted that a limit test can be performed with TG 414 (line 173), and that an extended one-generation study could replace the two-generation study in the near future (line 392). The section on alternative methods is comprehensive, but uncritical. It includes the amphibian FETAX assay (line 457), which did not impress in a validation study, and zebra fish embryo (line 438) and chick embryo (line 471) assays, which are unlikely to be necessary, if suitable mammalian stem cell-based tests (line 508), and especially, tests employing human material (lines 524 and 543), become available. It is surprising that tests based on human embryonic stem cells are not regarded more positively by the authors of this draft chapter, and it should be noted that the use of such cells in combination with metabolomics shows great promise for predicting human developmental toxicity (16). Many other in vitro tests and biochemical tests, involving, for example, the placenta, male fertility and endocrine disruption, are described, but their relevance to the safety of cosmetic ingredients is given little attention. After a brief review of their current status, it is concluded that "in silico approaches to predict reproductive toxicity are seldom robust and often have poor predictive capacity" (line 924). A brief section on QSARs and ADME related to reproductive toxicity (line 870) is blatantly focused on pharmaceuticals and oral dosing, and the need for in silico systems for predicting the uptake of cosmetic ingredients is not given the attention it surely deserves. The in vitro systems are criticised for representing "only a very simplified picture of reality" (line 973), but the limitations of the current strategies based on animal testing, are overlooked. That final point is a crucial one, as the draft chapter does not recognise the warnings given by Thomas Hartung (17), Hartung and Costanza Rovida (18) and others (e.g. 19, 20), concerning the cost of animal testing and its inadequacy, which threaten disaster for the REACH system, and, unavoidably, for any other toxicity or safety evaluation which relies on the current in vivo toxicity test procedures. This is so important that we think it is worth including a long quotation from Hartung's Tenth FRAME Annual Lecture (17): "The thalidomide scandal had a tremendous impact on toxicology. Between 1956 and 1962, approximately 10,000 children in Africa and Europe were born with severe malformations, because their mothers had taken thalidomide during pregnancy. As a result, ever since then, there has been a strong emphasis on testing to identify the teratogenic potentials of synthetic substances. Hence, more than 70% of the costs and more than 80% of the animals required for compliance with the EU REACH testing programme for new and existing chemicals will probably be devoted to reproductive toxicity testing. Although experts estimate that only 2% of birth defects are caused by chemicals (21), the belief that we must identify these, whatever it costs, is difficult to overcome. Nevertheless, as we have shown (22), all this effort, cost and animal use might actually result in the false-positive assignment of reproductive toxic potential to about 60% of all the substances tested! This is because reproductive toxic potential has a low prevalence among the thousands of chemicals to which the REACH system will apply, coupled with the low predictive value of the animal tests currently required by the regulators. Thus, the true positive reproductive toxicants might not be identified, because of a large number of false-positive results, and many valuable chemicals might be unnecessarily used only under strictly controlled conditions or not approved for use at all!" 352 Comment What is needed is a thorough and independent review of the purpose of reproductive toxicity testing and how it should be conducted, with a harmonised approach for chemicals and pharmaceuticals. The inescapable fact is that the predictive value of animal test data for humans is poor (23). The urgent need for new concepts has been put very powerfully in a recent authoritative comment on The way forward in reproductive/development toxicity testing (24), which is not listed among the 105 references attached to the draft chapter. Until such a review has been conducted, and a set of constructive proposals have been presented for critical evaluation, we support the proposal by Hartung and Rovida (18) for "a moratorium on reproductive-toxicity testing, or at least limiting testing to the most suspicious substances, until the OECD guidelines [for an extended one-generation study] are completed and alternative strategies for screening lots of chemicals are available". Meanwhile, since no convincing case has been presented for the reproductive toxicity testing of cosmetic ingredients as a special case for application in addition to the testing conducted in compliance with the REACH system, such testing should be banned forthwith, and should certainly not be conducted beyond 11 March 2013. Conclusions Without substantial revision, these five draft chapters cannot provide a credible basis for the Commission's report to the European Parliament and the European Council on the five cosmetic ingredients testing issues for which the ban on animal testing was postponed until 2013. Nor could they be used to provide reliable estimates on the time needed to develop and validate replacement alternative test methods. This is principally because the draft reports of the five groups of experts are of dubious relevance to any actions related to the Cosmetics Directive, because they are not sufficiently focused on issues related to the nature of cosmetic ingredients, their uses, their local effects and metabolism at their sites of application, and their possible uptake and absorption into the circulation, resulting in their distribution to visceral organs and tissues. Without meaningful predictions of the likelihood of uptake and, where uptake is likely, of the amounts that would be distributed to various internal sites, discussions on repeated dose testing, toxicokinetics, and, in particular, carcinogenicity and reproductive toxicity, can have little or no value as a basis for deciding what tests should be conducted and what tests could safely be banned. A crucial question is whether the uptake of a cosmetic ingredient could lead to a level in the body which approached the TTCs for various possible toxic effects. If, as would probably be the case in the vast majority of instances, such TTCs would be unlikely ever to be approached, then testing for the toxicity concerned, whether by in vivo, in silico or in vitro methods, could not be justified as necessary. The MoU concept, mentioned above, deserves serious consideration. Indeed, a different kind of risk assessment would be advisable, namely, an Absorption Risk Assessment, i.e. the risk that an ingredient and/or its metabolites might be absorbed and accumulate at levels which would approach TTCs and would therefore justify a Toxicity Risk Assessment. This whole exercise would have been much more useful, if the Commission had asked another working group of experts to consider the uptake of cosmetic ingredients, and to produce a report before the other working groups began their deliberations. It is especially disappointing that so little attention is paid in the draft chapters to intelligent testing strategies and/or integrated testing strategies. Many schemes have been proposed, which are related to the toxicity of chemicals and to pharmaceuticals, as evaluated in compliance with the REACH system legislation or directives other than the Cosmetics Directive. The leading cosmetic companies undoubtedly have such schemes for inhouse use, which must take account of dermal penetration or other routes of entry into the body. It would have been useful to have this reflected more fully in the draft chapters. It is particularly important that such schemes show how in silico and in vitro methods, along with the use of other information not derived from animal experiments, could be used in complementary and mutually supportive ways, to show how human welfare could best be protected, without resort to compromising animal welfare. It is not clear, from the introductory pages from the Commission, how the experts in the five working groups were nominated, and how they were selected. Apparently there was no open call for experts to serve on the working groups. There remains a lack of transparency, since the experts seem to have been "hand-selected", and the resulting evaluations are not unbiased. Of the 41 experts, 6 were from Commission's JRC, 7 from industry, 11 from research centres, 16 from academia, with one private consultant. Their geographic locations were as follows: Belgium (2), Denmark (1), Finland (4), France (1), Germany (8), Italy (8), Spain (1), Switzerland (1), The Netherlands (6), the UK (7), and the USA (2). Nor is there any indication of what information was given by the Commission to the working groups, to guide them on how their discussions on cosmetic ingredients should be conducted, bearing in mind the requirements of other legislation, and, in particular, that related to the REACH system. Comment 353 Decisions about the risk and safety of cosmetic products must surely depend on what is known about the chemicals they contain, most of which are likely to be used in other kinds of products. The Cosmetics Directive cannot be meaningfully applied in a knowledge vacuum. References 1. Anon. (2004). 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(2008). Toward a new toxicology -- evolution or revolution. ATLA 36, 635�639. 18. Hartung, T. & Rovida, C. (2009). Chemical regulators have overreached. Nature, London 460, 1080�1081. 19. Spielmann, H. & Vogel, R. (2006). REACH testing requirements must not be driven by reproductive toxicity testing in animals. ATLA 34, 365�366. 20. Combes, R. (2007). Reproductive toxicity testing under the REACH System: Time for a paradigm shift. ATLA 35, 1�4. 21. Schaefer, C., Spielmann, H. & Vetter, K. (2006). Arzneiverordnung in Schwangerschaft und Stillzeit, 781pp. Jena, Germany: Urban & Fischer. 22. Bremer, S., Pellizzer, C., Hoffmann, S., Seidle, T. & Hartung, T. (2007). The development of new concepts for reproductive toxicity applicable to large scale programmes. Current Pharmaceutical Design 13, 3047�3058. 23. Bailey, J., Knight, A. & Balcombe, J. (2005). The future of teratology is in vitro. Biogenic Amines 19, 97�145. 24. Spielmann, H. (2009). The way forward in reproductive/development toxicity testing. ATLA 37, 641�656. After this Comment was completed, the Editoriala for this issue of ATLA was received from Horst Spielmann, Associate Editor, Europe. It makes significant comments about the consultation and, in particular, about draft Chapter 5, on Reproductive Toxicity. a Spielmann, H. (2010). The EU Commission's Draft Report on Alternative (Non-animal) Methods for Cosmetics Testing. Current Status and Future Prospects -- 2010: A Missed Opportunity. ATLA 38, 339�343.