Source: EURLEX
Language: en
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###### **COMMISSION OF THE EUROPEAN COMMUNITIES**

Brussels, 05.05.1997
COM(97) 182 final

#### **1996 COMMISSION REPORT** **ON THE DEVELOPMENT, VALIDATION AND LEGAL** **ACCEPTANCE OF ALTERNATIVE METHODS TO ANIMAL** **EXPERIMENTS IN THE FIELD OF COSMETICS.**

###### TABLE OF CONTENTS **SUMMARY 4**

**PARTI** **8**

###### 1996 COMMISSION REPORT

**A. INTRODUCTION AND BACKGROUND** **8**

**B. OBJECTIVES AND CONSTRAINTS** **9**

1. Consumer safety 9

2. The reduction and elimination of animal suffering 10

3. International aspects and the GATT/TBT rules 12

4. The interests of the SMEs 14

**C. EXISTING AND EXPECTED RESULTS** **14**

1. Tests on ingredients and combinations of ingredients 14

2. Finished cosmetic products 16

**D. CONCLUSIONS DRAWN BY THE COMMISSION FROM THESE**
**OBJECTIVES, CONSTRAINTS AND RESULTS IN ITS DRAFT DIRECTIVE**
**PARTIALLY POSTPONING THE DEADLINE OF 1 JANUARY 1998 AND IN**

**ITS** **REFLECTIONS ON THE PRESENTATION OF A PROPOSAL FOR A**

**EUROPEAN PARLIAMENT AND COUNCIL DIRECTIVE AMENDING ARTICLE**
**4(1)(l) OF THE BASIC DIRECTIVE** **17**

**E. CONCLUSIONS AND OUTLOOK** **18**

**A**

**PART** **2** **21**

SCIENTIFIC ASPECTS

**A. INITIATIVES TAKEN** **IN** **1996** **21**

1. ECVAM 21

2. Scientific Committee on Cosmetology (managed by DG XXIV) 22

3. DG XII 22

4. COLIPA 23

5. OECD... 24

**B. STATISTICS** **ON** **ANIMAL EXPERIMENTS** **25**

**C. STATE** **OF** **PROGRESS IN** **1996** **IN** **EACH INDIVIDUAL AREA** **26**

1. Photoirritation/phototoxicity 26

2. Percutaneous absorption 27

3. Eye irritation 30

4. Skin irritation 30

5. Skin sensitisation 31

6. Skin compatibility of the finished product 32

### **`Summary`**

##### `INTRODUCTION AND BACKGROUND`

This 1996 report is the third report to be presented by the Commission. It reviews
the situation at October 1996, i.e. just before the date by which the Commission
had to present draft measures postponing the date for the ban on animal testing in
cases where alternative methods of testing have not been scientifically validated
as offering an equivalent level of protection for the consumer, taking into account
OECD guidelines. It belongs in the context of the sixth amendment to the basic
Cosmetic Products Directive adopted by the Council on 14 June 1993 (Directive
93/35/EEC), which provides for the banning of cosmetic products which contain
ingredients or combinations of ingredients tested on animals as of 1 January 1998
unless alternative methods are not available by this date, in which case the
Commission must present draft measures to postpone the ban. The sixth
amendment also provides that the Commission shall present annual progress
reports to the EP and Council.

The 1996 report consists of two parts. The first, more general part:

- describes the objectives and constraints which have to be addressed.

- summarises the existing and expected results,

- specifies the conclusions drawn by the Commission from these objectives,
constraints and results in preparing its draft proposal for a Directive
postponing the deadline of 1 January 1998.

The second part of the report is more scientific. It explains:

- the initiatives taken in 1996 by the various parties involved in the
research, validation and legal acceptance of alternative methods.

- the difficulties involved in obtaining statistics,

- the state of progress in each individual domain.

OBJECTIVES AND CONSTRAINTS

There are _two key objectives_ - namely ensuring the safety of cosmetic products
for human health and the elimination/reduction of animal suffering.

Cosmetic products, which include personal hygiene products, are used
throughout a person's life. The Cosmetic Products Directive makes it
incumbent on manufacturers to ensure that cosmetic products are safe for
human health and, as of 1 January 1997, to keep a safety information

dossier which will be accessible to the monitoring authorities. The
Cosmetic Products Directive has also established a system of lists of
substances that are banned, subject to restrictions, or authorised
respectively. These lists are regularly adapted to technical progress, on the
basis of an opinion delivered by the Scientific Committee on
Cosmetology, which takes into account the results of tests.

The banning of all new ingredients would not make tests superfluous
because the safety of ingredients must be regularly reexamined in the light
of scientific progress. Neither can one outlaw all innovation - a measure
which would bankrupt numerous SMEs.

- The reduction and elimination of animal suffering is an ethical imperative.
Although animal tests in the field of cosmetic products account for a mere
0.03% of all animal tests, they have been given priority.

The report describes the work done in 1996 by the following parties:

    - the Commission and notably ECVAM, the European Centre for the
Validation of Alternative Methods, a unit of the Joint Research
Centre; DG XI "Environment, Nuclear Safety and Civil Protection,
which manages Directive 86/609 on the protection of animals used
for scientific and experimental purposes; DG XII "Science,
Research and Development", in funding research programmes;
DG XXIV "Consumer Policy", which manages the Cosmetic
Products Directive and the Scientific Committee on Cosmetology.

    - the Scientific Committee on'Cosmetology;

   - COLIPA/the European Cosmetic, Toiletry and Perfumery
Association;

    - the OECD.

However, the report emphasises that the development, validation and acceptance
of alternative methods is a process which has proven more complex and arduous
than initially foreseen and that, as in all scientific research, it is impossible to
guarantee a precise result by a given date. The report also stresses that the utmost
prudence is called for as regards the use of human volunteers, who cannot serve
as a substitute for animal experiments pure and simple.

_The constraints_ that have to be borne in mind are mainly:

 - compliance with the rules of international trade, notably those of the WTO
(World Trade Organisation). The point is that any measure having the
effect of banning products from third countries on the grounds that these
products have been tested on animals poses problems of compatibility
with the rules of international trade. It would appear necessary to
investigate the subject in greater detail in the context of preparing an EP

and Council Directive which the Commission intends to propose during
the coming months, designed to amend the basic Cosmetic Products
Directive so as to address, in a legally appropriate text, the ban on animal
testing by regulating in particular the question of finished products.

the interests of the employment-generating SMEs, which also must be
taken into account. Measures must be taken to ensure that they are trained
and informed.

EXISTING AND EXPECTED RESULTS

The results are summarised in the first part of the report and described in greater
detail in the second part.

A distinction must be made between (a) ingredients and combinations of
ingredients and (b) finished cosmetic products.

As regards _tests on_ _ingredients/_ _combinations of ingredients_ the report makes it
clear that no alternative method offering an equivalent degree of protection is
available to date, or likely to be available by 1 January 1998.

Methods which do not involve animal trials will however become progressively
available in the fields of percutaneous absorption and local risks to the eye and
skin: photoirritation, eye irritation and skin irritation.

However, as regards tests concerning the system risks - i.e. risks involving
exposure of the organism as a whole as opposed to local risks - there is no hope
that animal trials can be replaced on schedule. However, the number of animals
used is being steadily reduced.

In general it will be possible to evaluate the safety of the _finished products_ after
1 January 1998 without recourse to animal trials, thanks notably to existing
knowledge on the safety of the ingredients, and by methods not involving the use
of animals, even if these methods are not liable to be the subject of an OECD
guideline, which deals exclusively with ingredients and combinations of
ingredients. However, in certain cases there may be a risk of toxic effects, due for
example to the interaction of the ingredients or to the fact that skin penetration of
the ingredients is facilitated by the vehicle used. In such cases tests on animals
would remain necessary.

CONCLUSIONS DRAWN BY THE COMMISSION FROM THESE

OBJECTIVES, CONSTRAINTS AND RESULTS

The Commission's draft directive designed to postpone the deadline of
1 January 1998 takes into account the objectives, constraints and results described
in the report.

The Commission notes that, while progress has been made in research into
alternative test methods, notably in the fields of percutaneous absorption and local
risks to the eyes and skin, the fact remains that no alternative method has yet been
scientifically validated and accepted andthat the OECD has not yet adopted
pertinent guidelines on toxicity tests in the field of alternative testing methods.

The situation is unlikely to change before 1 January 1998.

Hence the draft Directive proposes the postponement of the date mentioned in
Article 4(1 )(i) of Directive 76/768/EEC, in compliance with the second sentence
of that provision, until 30 June 2000, before which date it can be foreseen that no
alternative testing method will have been adequately scientifically validated and
accepted.

It also provides that before 1 January 2000 the Commission shall present draft
measures taking into account progress made by that date.

The Commission also wishes to promote research into and the validation of
alternative methods.

As regards finished products, the Commission will take measures to promote the
dissemination among SMEs of methods not involving the use of animals.

The Commission has also initiated reflections and intends, in the coming months,
to present a proposal for a European Parliament and Council Directive on the
amendment of Article 4 (1) (i) to address, in a legally appropriate text, the ban on
animal testing in respect of finished cosmetic products - barring exceptional cases

- as of 1 January 1998, in order to comply with the requirements of Directive
76/768.

This proposal for a Directive should also regulate aspects relating to products
from third countries and clarify certain concepts and references in the text that
have given rise to questions.

Since the Directive will be based on Article 100a, this Commission proposal
requires the approval of the European Parliament and the Council.

### **f**

**PART!**

### **1996 Commission Report**

on the development, validation and legal acceptance of alternative methods to animal

experiments in the field of cosmetics.

```
A. INTRODUCTION AND BACKGROUND

```

The Commission has already presented two annual reports on 1994 and 1995 data
concerning progress in the development, validation and legal acceptance of
alternative methods to animal experiments in the field of cosmetics.

This 1996 report reviews the scientific situation at October 1996.

This 1996 report belongs in the context of Directive 76/768/EEC on cosmetic
products, whose new Article 4 (1) (i), introduced by Council Directive 93/35 of
14 June 1993, provides that Member States shall ban the placing on the market of
cosmetic products containing ingredients or combinations of ingredients tested on
animals after 1 January 1998 in order to meet the requirements of this Directive.

However, this Article specifies that "if there has been insufficient progress in
developing satisfactory methods to replace animal testing, and in particular in
those cases where alternative methods of testing, despite all reasonable
endeavours, have not been scientifically validated as offering an equivalent level
of protection for the consumer, taking into account OECD toxicity test guidelines,
the Commission, shall, by 1 January 1997, submit draft measures to postpone the
date of implementation of this provision, for a sufficient period, and in any case
for no less than two years, in accordance with the procedure laid down in Article
10. Before submitting such measures, the Commission will consult the Scientific
Committee on Cosmetology".

This 1996 report comprises two parts. The first, more general part, describes the
objectives and constraints that have to be addressed, the existing and expected
results as regards ingredients and combinations of ingredients on the one hand
and finished products on the other, and the conclusions drawn by the Commission
from these objectives, constraints and results in preparing its draft proposal for a
Directive to postpone the deadline of 1 January 1998.

The second part of the report is more scientific. It describes:

the initiatives taken in 1996 by the different parties involved in the
development, validation and legal acceptance of alternative methods,
specifying wherever possible the cost of these initiatives

     the difficulties in obtaining statistics

**8**

the state of progress in each individual domain.

**B.** **OBJECTIVES AND CONSTRAINTS**

Two principal objectives must be pursued and wherever possible reconciled in the
field of cosmetic products: (a) ensuring consumer safety and (b) reducing and,
ideally, eliminating animal suffering.

Moreover, various constraints must be borne in mind, the two primary ones being
(a) the international arena and the need to comply with WTO rules and the rules
of free competition, and (b) the interests of firms, notably employment-generating
small and medium-sized enterprises.

**1.** **Consumer safety**

Consumer safety must remain paramount. Here it should be noted that the
notion of "cosmetic products" encompasses not only so-called
"decorative" products, such as lipstick or nail varnish, but also articles
such as soap, shampoos, toothpaste which are used throughout a person's
lifetime from early childhood on. It is essential that these products should
not cause harmful, immediate and visible effects, such as irritation, or
long-term, latent effects such as risks of carcinogenicity or teratogenicity,

etc.

To ensure that cosmetic products are safe for human health in all these
respects, it is necessary to check not only the safety of the finished product
but also - and primarily - that of the ingredients used in its manufacture.
It is for the manufacturer to ensure the safety of the ingredients, in
compliance with Article 2 of the Cosmetic Products Directive, according
to which cosmetic products must not be liable to cause damage to human
health when they are applied under normal conditions of use;
manufacturers may also be held liable in the event of damages pursuant to
Council Directive 85/374/EEC on liability for defective products.
Pursuant to Council Directive 93/35/EEC of 14 June 1993 amending for
the sixth time the basic Cosmetic Products Directive, the manufacturer
must, as of 1 January 1995, keep certain information readily accessible to
the competent authorities, including "assessment of the safety for human
health of the finished product. To that end the manufacturer shall take into
consideration the ' general toxicological profile of the ingredient, its
chemical structure and its level of exposure."

The Cosmetic Products Directive also includes a series of annexes

featuring positive and negative lists of substances. The general principle is
that substances may be used freely, unless they are prohibited (Annex II),
or subject to certain limitations and conditions (Annex III). There are three
exceptions to this general principle: the only colouring agents,
preservatives and UV filters authorised are those featuring in positive lists
(Annexes IV, VI and VII).

A substance can only be included in these lists, which are adapted to
technical progress each year by a Commission Directive, if it has been the
subject of an opinion delivered by the Scientific Committee on
Cosmetology concerning its toxicity for human health. This SCC opinion
is prepared on the basis of all the existing scientific data and notably the
test results furnished by industry, which may concern sensitisation,
mutagenicity, eye irritation, skin irritation, photoirritation,
carcinogenicity, teratogenicity, and acute, subchronic and chronic toxicity,

etc.

While some 7 000 substances are already used in cosmetic products and
while for many cosmetic substances new tests are not generally required.
it should however be stressed that such tests may sometimes be crucial in
reevaluating, on the basis of technical progress and new scientific
knowledge, certain substances which have been in use for a very long
time. Hence each year the SCC examines a series of banned substances
(numbering approximately 400) and substances subject to certain
limitations and requirements (numbering approximately 50).

It must also be possible to examine the risks associated with the use of
these products in combination. Thus on safety grounds it is not possible to
dispense entirely with animal trials, even in respect of substances already
in use. Moreover, in the cosmetic products sector - as in all other
industrial sectors - it would be neither wise nor reasonable to bring
innovation to a standstill. Even if consumers were to foresake their quite
legitimate hope of obtaining ever better products, a blanket ban on
innovation by outlawing the use of new substances would certainly spell
ruin for numerous European cosmetic product manufacturers, mainly
employment-generating SMEs.

**The reduction and elimination of animal suffering**

Reduction and, where possible, elimination of animal suffering is an
objective that deserves the closest attention and in respect of which
resources should be mobilised at all levels.

This objective reflects ethical imperatives concerning respect for life,
deep-seated public aspirations, and the wishes of the European Parliament.
It finds expression in several European instruments, including Directive
86/609/EEC on the protection of animals used for experimental and other
scientific purposes and Directive 93/35/EEC (the sixth amendment to the
initial Cosmetic Products Directive).

While consumer safety must be assured, we must also do our utmost to
ensure that animal tests are replaced by alternative tests as soon as
possible, provided these afford an equivalent level of protection. Although
animal trials in the cosmetics field account for a mere 0.03% of all animal

tests and although all chemical substances already have to be tested in

**10**

order to provide the toxicity data required under Directive 92/32/EEC,
cosmetic products have been prioritised and there has been a major drive
in recent years, notably since the adoption of Directive 93/35/EEC, to
promote the development of alternative methods. These endeavours, as
well as progress to date and the outlook for the future, are described in the
1994 and 1995 annual reports on the development, validation and legal
acceptance of alternative methods to animal experiments in the field of
cosmetic products, already presented by the Commission to the EP and the
Council.

Notably, these reports describe the work done by a number of players. Part
II reviews the activities of these players in 1996.

In particular, these players are:

_ECVAM_ (the European Centre for the Validation of Alternative Methods),
a unit of the Ispra Joint Research Centre, which coordinates the validation
of alternative methods at Community level, provides a forum for pooling
information and works on the development, updating and management of
a database and promotes dialogue between all interested parties.

In 1996 _ECVAM_ continued to organise workshops of direct relevance to
evaluating the safety of cosmetic products (based on e.g. data concerning
alternative methods); it published in ATLA recommendations methods for
assessing percutaneous absorption and for skin sensitisation testing, and
recommendations on the application of biostatistical methods to the
development and validation of alternative toxicological methods; it
subsidised and coordinated a study for testing UV filters listed in Annex
VII to the Cosmetic Products Directive (photoirritation); it conducted a
formal validation study on four tests of skin corrosion, and maintained
regular contacts with the OECD and began a number of prevalidation
studies relevant to the cosmetics industry.

The Commission _(DG XI_ "Environment, Nuclear Safety and Civil
Protection") which manages Directive 86/609/EEC on the protection of
animals used for experimental or other scientific purposes, has continued
to follow the matter. The Commission _(DG XII_ "Science, Research and
Development") has also subsidised research programmes including the
development of alternative methods.

In the context of its consumer policy, the Commission _(DG XXIV)_ has
funded research and participated in or organised meetings with all the
partners involved and organised and followed up the meetings of the SCC.

_The Scientific Committee on Cosmetology_ - whose mandate is to deliver
scientific opinions to the Commission on ingredients listed in the Annexes
to the Cosmetic Products Directive and on the applicability of alternative
methods validated in the process of evaluating the safety of cosmetic
products, and on the proposal for a Directive which the Commission must

**i l**

present concerning the postponement of the deadline for the ban on animal
tests in cases in which alternative methods will not be available by
1 January 1998     - met on numerous occasions in 1996 both in plenary and
in its Alternative Methods Subcommittee, sometimes jointly with the
European Industry. Besides the activities documented in Part II, the SCC
adopted in 1996 a document titled "The use of alternatives to animal
studies in the safety evaluation of cosmetic ingredients or combinations of
ingredients".

Finally, at _OECD_ level, work has been done on percutaneous absorption
(meeting of the national coordinators in Paris in September 1996).

It is important to note that the development, validation and acceptance of
alternative methods cannot be achieved overnight, despite all the effort
that is being invested.

Hence the validation phases - i.e. the complex process by which the
pertinence, reliability and reproducibility of a test developed for routine
application and legal acceptance is evaluated and monitored - have turned
out to be more numerous and complex than initially foreseen, and new
phases have had to be incorporated.

Moreover, as in the case of all scientific research, it is well-nigh
impossible to guarantee a precise result by a given date, and certain
studies, which were thought to be on the verge of completion - such as the
EU/HO study on alternatives to the Draize test - turned out to be
disappointing as regards the predictability of the risk.

As regards alternative tests on human volunteers, for example in the field
of skin irritation, the utmost prudence is called for; such tests should only
be countenanced in the proven absence of real risks.

For all that, progress to date has already made it possible to substantially
reduce the number of animals used and to mitigate their suffering. - for
example the use of three basic in vitro mutagenicity tests will obviously
lead to further improvements in the future.

3. **International aspects and the** **GATT/TBT** **rules**

It is important to note that any measure that leads to banning the
importation of cosmetic products into the Community solely because these
products or their ingredients have been tested on animals might prove
incompatible, with the rules of international trade. The main restriction that
public international law imposes on a state is to forbid - except in the
event of a specific rule to the contrary - any exercise of its power on the
territory of another state.

**12**

Firstly, the WTO outlaws quantitative restrictions on imports (Art XI§1).
Moreover, Article III §4 proscribes all discriminatory measures between
_like_ products, i.e. in accordance with the accepted interpretation, products
which do not present physical differences. Since the test method clearly
has no physical effect on the product, a discrimination based on this
criterion might be held to fall foul of Article III§4.

Finally, such a measure would not be easy to justify in the context of the
general exceptions featured in Article XX of the GATT (and more
specifically subsection b), which authorises - within the terms of Article
XX - measures "necessary to protect human, animal or plant life or
health". In the absence of case law in this area, it is not possible to claim
that protection of animal welfare could be covered by the exception.

Secondly, as regards measures designed to verify compliance with the
provisions of the Cosmetic Products Directive - viz. requirements in
regard to the protection of human health - the provisions of the Agreement
on Technical Barriers to Trade (TBT) and notably those of Article 6 must
be borne in mind. Article 6 promotes the mutual recognition of conformity
evaluation procedures: "Members shall ensure that, whenever possible,
that results of conformity assessment procedures in other Members are
accepted, even when these procedures _differ_ from their own, provided they
are satisfied that those procedures offer an assurance of conformity with
applicable technical regulations or standards _equivalent_ to their own
procedures".

Bearing these provisions in mind, the extraterritorial application of a
Community measure banning animal'trials could be considered as contrary
to the Community's international commitments. Moreover, Article 4(1 )(i)
of this Directive specifies that the ban on animal testings may be
postponed if there are no scientifically validated alternative methods
which offer the consumer an _equivalent_ degree of protection. Hence it
would be difficult to deny the equivalence, from the viewpoint of human
health protection, of animal tests with alternative test methods which are
recognised as equivalent.

It should be noted that the rationale behind the ban is not that animal tests

do not permit correct assessment of compliance with the provisions of the
Cosmetic Products Directive, but is based purely on ethical
considerations.

Thus any decision on the use of alternative methods could also be
challenged if it were adopted on a unilateral basis. Moreover the TBT
Agreement strongly encourages the members of the WTO to adopt the
pertinent international systems for assessment of the conformity of a
technical regulation (Article 9). However, Article 9 also specifies that this
applies only to the extent that these international systems are in
compliance with the provisions of Articles 5 and 6 of the selfsame TBT
Agreement (Article 9§3).

Thus it is very important to remain at least within the framework of the
Organisation for Economic Cooperation and Development (OECD), to
which moreover Directive 93/35 refers.

Part II of this report explains the role played by the OECD in regard to
alternative methods in the different domains of the safety of cosmetic
products and highlights the close links which the Commission has also
developed and maintains with the OECD in this specific field.

The Commission is reflecting on to all matters relating to the rules of
international trade in the context of preparing a proposal for a European
Parliament and Council Directive designed to amend the basic Cosmetic
Products Directive to address, in a legally appropriate text, the ban on
animal testing, notably by regulating the question of finished cosmetic
products.

**4.** **The interests of the SMEs**

Finally, apart from consumer safety and the ethical imperative to
reduce/eliminate animal suffering, the reasonable interests of the
cosmetics industry should also be taken into consideration, notably those
of the employment-generating SMEs. The alternative methods to be
developed should be accessible to SMEs, even if they do not have as many
"in-house" toxicologists or the same amount of experience or field data, or
the internal laboratories and financial resources available to the large
firms.

In this context, COLIPA will be launching a major drive to provide
information and training, in collaboration with its large member firms.
Moreover, it seems that specialised laboratories are springing up whose
services the SMEs will be able to draw on for alternative trials, as and
when such trials reach maturity.

ECVAM could also be involved - possibly together with COLIPA (Scaat)

       - in training SMEs in the use of alternative methods for evaluating the
safety of cosmetic products.

**C.** **EXISTING AND EXPECTED RESULTS**

**1.** **Tests on ingredients and combinations of ingredients**

It should be noted that, as shown in Part II, no alternative method offering
consumers an equivalent degree of protection has yet been scientifically
validated and accepted by the regulatory authorities. The OECD has not
yet adopted pertinent guidelines for toxicity tests in the field of alternative
testing methods for ingredients and combinations of ingredients used in
cosmetic products.

**14**

However progress has been made and it looks as though alternative
methods will progressively become available in the various safety
domains pertaining to percutaneous absorption and local risks to the eye
and skin (photoirritation, eye irritation and skin irritation).

However, these methods will not be validated _and accepted_ by 1 January
1998, despite the high expectations that were had, mainly in regard to
percutaneous absorption and photoirritation.

In regard to percutaneous absorption and local risks, the situation is as
follows:

_Photoirritation_

Photoirritation mainly concerns UV filters. A special study to follow the
EU/COLIPA validation study is scheduled for 1996. This is directed by
Zebet, and subsidised and coordinated by ECVAM. The study should
make it possible to test UV filters in Annex VII to the Cosmetic Products
Directive and to prepare a protocol with a view to submitting a draft
guideline to the OECD. The first results of this special study should be
available by September 1997.

_Percutaneous absorption_

In May 1996, after discussions with the Commission, the industry
presented the OECD with a draft new guideline on in vitro percutaneous
absorption. "In-house" data were provided to support this dossier. A
meeting of the OECD national coordinators' group was held in Paris in
September 1996, but the rate of progress toward acceptance slow.

The Scientific Committee on Cosmetology has emphasised that the test
protocols used by industry had not yet been subjected to the formal
validation study and that the documentation was incomplete. However, the
SCC has in recent years agreed to use in vitro percutaneous absorption
data in evaluating the safety of several cosmetic ingredients.

_Eye irritation_

The 1995 Report stressed that the results of the validation studies of
alternative methods to the Draize eye irritancy test had been disappointing
(EU/Home Office study and COLIPA study).

ECVAM and COLIPA held a meeting in December 1996 to discuss the
outcomes of the EC/HO and COLIPA studies and plan new initiatives.

**15**

_Skin irritation_

ECVAM has conducted a formal validation study involving four skin
corrosion tests, whose results are expected in 1997.

The OECD is also planning a draft testing strategy to determine the skin
irritation potential of chemical substances.

The draft OECD strategy is to use a hierarchical approach by eliminating
severe skin irritants through consideration of the structure activity
relations (SAR's) and structure property relations (SPR's) and data
obtained from in vitro corrosion tests, before considering the use of
conventional in vivo tests on rabbits or the use of human volunteers.

A summary of the situation regarding local risks shows that progress
should proceed apace but that 1 January 1998 is too early for the
replacement of animal trials. However, as regards reducing the number of
animals and their suffering, it may be noted that in practice in vitro tests
are already used as screening or guideline tests and that certain data have
already been provided to the SCC.

_Skin sensitisation_

Research has led to a better understanding of the scientific bases of
skin sensitisation.

ECVAM is to take action in the light of the report of the ECVAM
workshop on this topic.

_Mutagenicity_

The combination of three basic in vitro tests already makes it
possible to demonstrate a mutagenic potential and this combination
is already being applied in evaluating the safety of cosmetic
ingredients.

_All tests on ingredients relating to systemic effects_

(Acute toxicity, subchronic toxicity, carcinogenicity,
teratogenicity). Because they are linked to exposure of the
organism as a whole, there is no hope that alternative
methods to replace animal trials will be available in the
foreseeable future.

However, the refinement of the methods used has already
made it possible to reduce the number of animals used and
mitigate their suffering, and this trend is likely to continue.

**2.** **Finished cosmetic products**

**16**

As regards finished cosmetic products, progress to date suggests that it
will generally be possible to evaluate their safety on the basis of available
knowledge on the toxicity of their ingredients and their physicochemical
properties, using methods which do not involve the use of animals, even if
these methods are not liable to be the subject of an OECD guideline which
deals dealing only with ingredients and combinations of ingredients.

Hence it will only be necessary to resort to animal testing in rare,
exceptional cases, where there are grounds to fear that the toxic effects of
the ingredients may be potentiated, notably when skin penetration of the
ingredients is facilitated by the vehicle used or when such effects result
from the interaction of the ingredients. The SCC has emphasised this fact
in its Guidelines on the use of alternative methods to animal studies in the

safety evaluation of cosmetic ingredients or combinations of ingredients of
24 Mav 1996.

Certain cosmetics firms already very often use methods which do not
involve the use of animals to test their finished products.

COLIPA (the European Cosmetic, Toiletry and Perfumery Association)
has announced that it could progressively establish a voluntary ban on
animal testing for finished cosmetic products - except in the exceptional
cases referred to above - for the entire European cosmetics industry, and
that it would be willing to organise technology transfer in this respect.
COLIPA has also said that it could ensure the widest possible
dissemination of findings in this area and promote training in European
companies and laboratories; this training could be coordinated by the
Commission's services. The Commission, in particular, will promote the
dissemination of methods not involving animals among the SMEs. It
could also endeavour to promote such measures at international level.

D. CONCLUSIONS DRAWN BY THE COMMISSION FROM THESE OBJECTIVES,

CONSTRAINTS AND RESULTS IN ITS DRAFT DIRECTIVE PARTIALLY POSTPONING

THE DEADLINE OF 1 JANUARY 1998 AND IN ITS REFLECTIONS ON THE

PRESENTATION OF A PROPOSAL FOR A EUROPEAN PARLIAMENT AND COUNCIL

DIRECTIVE AMENDING ARTICLE 4(1)(I) OF THE BASIC DIRECTIVE

1. The draft Directive prepared by the Commission takes account of the fact
that, while progress has been made in research into alternative test methods,
notably in the fields of percutaneous absorption and local risks to the eyes and
skin, the fact remains that no alternative method has yet been scientifically
validated and accepted. The OECD has not yet adopted pertinent guidelines for
toxicity tests in the field of alternative testing methods.

The situation is unlikely to change before 1 January 1998.

**17**

Hence the draft Directive proposes the postponement of the date mentioned in
Article 4(1 )(i) of Directive 76/768/EEC, in compliance with the second sentence
of that provision, for not less than two years.

Currently it is difficult to predict precisely when certain alternative methods for
testing certain ingredients or combination of ingredients will be scientifically
validated. However as it is likely that alternative methods will progressively
become available in the field of percutaneous absorption, photoirritation, eye
irritation skin irritation and skin sensitivation, and whereas, besides, the
Commission is mindful of the objective of Article 4(1 )(i), the draft Directive
postpones the deadline to a date by which it can be foreseen no alternative method
will have been adequately scientifically validated and accepted, viz. 30 June 2000.
It also stipulates that before 1 January 2000 the Commission shall present draft
measures taking into account progress made by that date.

Finally, in the context described above, and also in compliance with the
provisions of Article 130f(3) of the Treaty and Fourth Research Framework
Programme, the Commission will take the necessary measures to promote
research into and the validation of alternative methods to animal testing in the
field of ingredients and combinations of ingredients in cosmetic product
formulations. This could take the form of a recital in the draft proposal for a
Directive.

2. Presentation of a proposal for a European Parliament and Council
Directive amending Article 4(1 )(i) of the basic Directive

In the context described in point C.2. above relating to the existing and expected
results for finished cosmetic products, the Commission has initiated reflections
and intends, in the coming months, to present a proposal for a European
Parliament and Council Directive amending Article 4 (1) (i) in order to address, in
a legally appropriate text, the ban on animal testing in regard to finished cosmetic
products - barring exceptional cases - as of 1.1.1998, in order to respect the
obligations of Directive 76/768.

This proposal for a Directive should also regulate, in a legally appropriate text,
aspects concerning products from third countries, and clarify the concept of
"finished products".

Since the Directive will be based on Article 100a, this Commission proposal
requires the approval of the Council and the European Parliament.

E. CONCLUSIONS AND OUTLOOK

1. This report reviews the situation in the immediate run-up to the date by
which the Commission must present a draft measure postponing the
deadline of 1 January 1998 in areas in which alternative methods for
testing ingredients and combinations of ingredients will not be available
by that date.

2. Two objectives must be pursued: consumer safety, which the Commission
insists must not be compromised in any way, and the
elimination/reduction of animal suffering.

    - Banning all new ingredients would not remove the need for animal
tests because the safety of ingredients has to be reexamined in the
light of new scientific knowledge. Moreover, blocking all
innovation could put firms out of business and in particular the
employment-generating SMEs. It should be noted also that
cosmetic products include personal hygiene products, products for
babies (...) and not only so-called "decorative" products.

    - Priority should be given to eliminating animal suffering in the field
of cosmetic products, even if tests in this domain account for a
mere 0.03% of all tests. The Commission (ECVAM, DG XI, XII,
XXIV), the Scientific Committee on Cosmetology, the OECD and
Industry continued to work to this end in 1996.

However the validation process has turned out to be more complex than foreseen.
In the SCC's opinion the utmost prudence is called for as regards the use of
human volunteers, who cannot simply serve as a stand-in for animal experiments.

**j .** Two constraints must also be borne in mind:

a) compliance with the rules of international trade, notably those of the
WTO(World Trade Organisation). The point is that any measure having
the effect of banning products from third countries on the grounds that
these products have been tested on animals poses problems of
compatibility with the rules of international trade. It would appear
necessary to investigate the subject in greater detail in the context of
preparing an EP and Council Directive which the Commission intends to
propose during the coming months, designed to amend the basic Cosmetic
Products Directive so as to address, in a legally appropriate text, the ban
on animal testing by regulating in particular the question of finished
products.

b) the interests of the SMEs, which must be trained and informed.

k As regards the existing and expected results, a distinction must be made
between ingredients/ combinations of ingredients and finished products.

a) As regards ingredients and combinations of ingredients, no
alternative method will be ready by 1 January 1998. However,
methods should progressively become available for percutaneous
absorption, photoirritation, eye irritation skin irritation and skin
sensitivation.

**19**

However, in the case of tests concerning system risks, i.e. exposure
of the entire organism, there is no hope for alternative methods
maturing in the foreseeable future.

b) In general, it should be possible to ensure the safety of finished
products, thanks mainly to our knowledge of the ingredients,
without resorting to animal testing, after 1 January 1998. However,
the SCC has emphasised the need for exemptions to this rule in the
case of toxic effects caused, for example, by interactions of the
ingredients.

c) Progress has however already made it possible to reduce the
number of animals used.

5. The Commission has drawn conclusions from these objectives, constraints
and expected results:

     - the Commission has prepared a draft Directive postponing the date
of 1 January 1998 in regard to ingredients and combinations of
ingredients.

The Commission has also decided to pursue its deliberations on presenting
a proposal for a European Parliament and Council Directive amending
Article 4 (1) (i) of the basic Directive in order to address the ban on
animal testing, notably by regulating the question of finished products.

**20**

PART 2

**SCIENTIFIC ASPECTS**

**A.** **INITIATIVES TAKEN IN 1996**

**1.** **ECVAM**

Of the workshops organised and/or published by ECVAM in 1995 and
1996 on the development and validation of alternative tests, those of direct
interest to the assessment of consumer safety are as follows:

Workshop 13. Methods for Assessing Percutaneous
Absorption
Angera, Italy; 30 May-3 June 1994
Howes et al. (1996) ATLA 24, 81
106

Workshop 16 Acute toxicity testing in vitro and
the classification and labelling of
chemicals

Angera, Italy; 18-22 April 1994
Seibert et al. (1996) ATLA 24, 499
510

Workshop 19 Alternative methods for skin
sensitisation testing
Angera, Italy; 24-28 April 1995
de Silva et al. (1996) ATLA 24, 683705

Workshop 24 The development and validation of
expert systems for predicting toxicity
Angera, Italy; 1-4 October 1996
Dearden et al. (1997) ATLA 24, in

press

Workshop 25 Current Status and Future
Development of Databases on
Alternative Methods

Neublberg, Germany; 12-15
September 1996. Report in
preparation.

**21**

ECVAM has also published in ATLA recommendations on the application
of biostatistical methods in the development and validation of alternative
toxicological methods (ATLA 24, 511-530, 1996).

Of the other ECVAM Task Forces, those dealing with Prevalidation, Skin
Irritation and Integrated Testing are also particularly relevant to the
cosmetic testing issue.

In addition to the ECVAM validation studies on skin corrosivitv and

phototoxicity, prevalidation studies were begun on modified protocols for
the bovine corneal capacity and permeability (BCOP) test and the
fluorescein leakage (FL) test.

**2.** **Scientific Committee on Cosmetology (managed by DG XXIV)**

The Alternative Methods/Guidelines Subcommittee of the SCC organised
three joint meetings with COLIPA/SCAAT (15 February 1996,
19 March 1996, 24 April 1996), a meeting with the Director of ECVAM
(6 March 1996) and a meeting designed to draw conclusions and adopt an
opinion (6 May 1996), following consultations with DG XXIV.

During these meetings the following points were addressed:

a) Preparation of a discussion document on the "use of alternative
methods to animal studies in the safety evaluation of cosmetic
ingredients or combinations of ingredients.

b) Examination of the final report on the EC/HO study on the
validation of the Draize eye irritancy test.

c) Examination of the statistical assessment data of phase 1 of the
COLIPA study - validation of the eye irritancy test in the rabbit.

d) Discussion with ECVAM's Unit head on the implementation of the
provisions of Article 4 of Directive 93/35/EEC. involving a
general review of the validation studies.

e) Discussions with COLIPA/SCAAT on Industry's approach to the
implementation of Directive 93/35/EEC, which is based on the
experience of the cosmetic firms and the outlook for replacing
animal tests.

3. DG **XII**

Since 1985 the Commission has been supporting research in the field of
new in vitro alternatives to animal experiments in pharmaco-toxicology
applicable to the cosmetic industry.

The BIOTECH II programme launched after the BAP, BRIDGE and
BIOTECH I programmes is also contributing to the development of these
in-vitro alternatives applicable to cosmetic products. Following the second
call for proposals, three projects are being funded with a total budget of
ECU 3 825 000.

The projects are: .

      - BIO4-CT96-0086:

New immuno-pharmaco-toxicological model: human
reconstructed epidermis containing Langerhans cells
Coordinator: Dr. Rainer Schmidt

L'OREAL S.A.

Centre Charles Zviak

9a, rue du Général Roguet
F - 92383 Clichy Cedex

      - BIO4-CT96-0036:

Development of _in_ _vitro_ systems using human
immortalized cell lines for testing skin irritancy
Coordinator: Professor Charles Lapière

Laboratory of Experimental Dermatology
Université de Liège
Tour de Pathologie, B 23
B - 4000 Liège

      - BIO4-C96-0246:

Development for _in vitro_ tests for drug allergenicity and B
cell switching to IgE synthesis
Coordinator: Dr. I. W. Coleman

Department of Pharmacology &
Therapeutics
New Medical Building
Ashton Street

GB - Liverpool L69 3BX

**4.** **COLIPA**

_COLIPA/SCAAT_ (the European Cosmetic, Toiletry and Perfumery
Association/ Steering Committee on Alternatives to Animal Testing)

Following the international two-day scientific symposium organised at the
end of 1995, Scaat has continued its efforts towards getting validated
alternative methods accepted and introduced into the regulatory
framework.

On EYE IRRITATION, the results of the COLIPA validation study
became fully available and were submitted for review to ECVAM and to
the SCC and DG XXIV. The relevant working party has drafted a first
paper on the outcome of the study, which has been accepted for

**23**

publication in _Toxicology In Vitro._ Further statistical analysis of the data
generated by the study is in progress.

On SKIN IRRITATION, the specific working party is drafting guidelines
on human testing of cosmetic ingredients, expected to be available early
1997. This follows the issuing of guidelines on human testing of finished
products, issued in 1995 and now published in _Food and Chemical_
_Toxicology._ Both guidelines incorporate the need for strict ethical
standards for such testing.

On PERCUTANEOUS ABSORPTION, a working party has been
collecting data generated by the cosmetic industry substantiating the
validity of in vitro testing, as laid down in the COLIPA guidelines
prepared in 1995. An extensive data package has been made available to
ECVAM, to OECD, and to the SCC and DG XXIV.

On PHOTOIRRITATION, a second phase of the EU/COLIPA validation
study has been successfully completed and discussed with the SCC. The
SCC has asked for a special study to verify the applicability of the in vitro
method for UV-filters listed in Annex VII of the Cosmetics Directive.

This special study will be sponsored by ECVAM and carried out over the

next year.

SCAAT has also created a working party "SKIN SENSITISATION". The
main objective of this new group is to define and execute a research
programme on mechanisms, which could pave the way to suitable in vitro
test methods.

5. **OECD**

OECD Guidelines for the Testing of Chemicals and the Principles of
Good Laboratory Practice (GLP) are developed in the broader context of
the concept of mutual acceptance of data. Both of these instruments for
ensuring harmonized data generation and data quality are an integral part
of the 1981 OECD Council Decision on the Mutual Acceptance of Data
(MAD). In accordance with this decision, OECD's 28 Member Countries
agree that data generated in the testing of chemicals in any OECD
Member Country, when in accordance with OECD Test Guidelines and
Principles of GLP, shall be accepted in any other Member Country for the
purpose of assessment and other uses relating to the protection of man and
the environment. The practical consequence of this decision is that data
developed in a Member Country under these conditions and submitted for
fulfilling regulatory requirements in another Member Country, cannot and
will not be refused. Consequently, OECD Test Guidelines are globally
accepted as the standard methods for safety testing and as such enhance
the validity and international acceptance of test data. Recognising the
significance of OECD Test Guidelines, the European Commission
strongly supports work on the development of new test guidelines and the
updating of existing ones.

**24**

**B.** **S T A T I S T I C S O N A N I M A L E X P E R I M E N T S**

On numerous occasions the Commission has urged the Member States, both
orally and in writing, to provide the necessary statistics.

1995:

1. Eight Member States reported that finished cosmetic products had not
been tested on animals on their territory (Italy, Greece, Belgium, Ireland,
Sweden, Finland, Luxembourg, Germany).

2. Six Member States reported that cosmetic ingredients had not been tested
on animals on their territory (Greece, the Netherlands, Ireland. Sweden,
Finland, Luxembourg).

3. Only three Member States (Austria, France, the United Kingdom)
communicated figures on the number of animals used, while emphasising
that these figures were not interprétable and that there was uncertainty as
to what they really signified. In these circumstances the 1995 Report could
not present comparative tables comprising numerical data, because such a
table would have given a false picture of reality.

4. As regards the declarations on thé absence of tests, it is worth mentioning
that certain Member States that do not manufacture ingredients will
certainly be using ingredients tested in other Member States or in third
countries and that certain ingredients used in cosmetic products may have
been tested for other purposes.

Following the final reminder from the Commission in October 1996:

     - Ireland and Finland confirmed that cosmetic products had not been tested
on animals in their countries in 1996. Sweden did likewise for 1995 but

reported that in November 1996 it did not have data relating to 1996.

     - Portugal reported that cosmetic products were not tested on animals in its

country.

     - Germany confirmed that finished products were not tested on animals and
that it had no data as regards ingredients testing.

     - Finally, in the Netherlands the Experiments on Animals Act, which
entered into force on 5 February 1997, provides that animal experiments
may not be carried out with a view to developing new cosmetics or to
testing existing cosmetics which come within the remit of the
Commodities Act.

The Commission regrets that it does not have more statistics and is pursuing its

efforts to obtain them.

**25**

**C.** STATE **OF PROGRESS IN 1996 IN** **EACH** **INDIVIDUAL** **AREA**

**1.** **Photoirritation/phototoxicity**

_ECVAM/COLIPA_

The biométrie analysis (= statistical evaluation) of the data produced
during the second validation phase has been completed for the 3T3 NRU
PI [1] cell viability test.

The raw data were imported into an EXCEL file and checked for internal
consistency. A new mathematical model was developed to analyse the
complex dose-response relationships.

The predictive value was evaluated by comparing the in vivo
photoirritation classification of the materials tested with the classification
based on the values and biological endpoint of the in vitro test.

The variability of the in vitro classification was calculated on the basis of
the rate of classification errors.

The degree of association between the results of the tests conducted by the
different laboratories was subjected to statistical analysis.

The results of the biométrie analysis (dated 29° March 1996) **indicate**
**that the NRU assay is capable of correctly predicting the irritation**
**potential in man** in regard to most of the substances tested. It may be
noted that:

a. (In many cases) the PIF does not represent the exactly measured
values but estimates which are below or above the critical value

(>5<) established in phase I of the validation study.

b. The prediction model proposed in the SOP (Standard Operation
Procedure) of the 3T3 NRU test predicts well the in vivo
phototoxicity of most of the substances tested.

c. The variability of the PIF values grows with the increase in
absolute PIF values.

The number of independent tests in each laboratory was too small
to authorise a valid estimate of intralaboratory variability data;

NRU: Neutral red uptake assay", which uses the cells as target and cell toxicity as the outcome.

26

d. The effects of data variability on the in vitro classification are
negligible, as the computer analysis of the classification errors
demonstrates.

The technical problems responsible for the incorrect classification of
certain test materials have been identified and will be taken into account in

evaluating the performance of the 3T3 NRU PI test in the final report.

In particular, one should bear in mind:

      - possible in vivo classification errors
errors in preparing the solution due to the total lack of information
on the coded test substances

      - the deviations in regard to the SOP.

_SCC_

**Following an SCC recommendation,** which emphasised the importance
of verifying the applicability of the NRU test in evaluating the safety of
Annex VII UV filters and creating a database, **a third experimental**
**phase,** _directed by_ _ZEBET,_ _and subsidised and_ _coordinated_ _by_ _ECVAM,_ is
**planned for** **1996.** The objectives of this study are **to test the Annex VII**
**UV filters** and to prepare an optimised protocol with a view to **submitting**
**a draft guideline on in vitro photoirritation to the OECD. The first**
**results** of the third phase **should be in by September 1997.**

_OECD_

The OECD group of national coordinators has decided not to develop the
in vivo method on its schedule before termination of the third in vitro

validation phase at end 1997. If the in vitro method has not been
completed by that date, the OECD will try to develop the in vivo method.

2. **Percutaneous absorption**

_COLIPA_

In May 1996 a method proposed in the ECVAM workshop report, as
discussed by the industry, ECVAM and OECD secretary, was presented to
the OECD.

This draft, which is currently being studied by the OECD group of
national coordinators, explicates the general principles governing the
design of protocols for skin penetration tests using the excised skin of
different mammalian species post mortem. The skin is excised from the
dead animals:

[- "know-how" regarding the substance tested and choice of vehicle;

**27**

choice of the type of cell, receptor fluid, cutaneous membrane and
control of membrane integrity;
potential significance of the skin metabolism;

- (finite or infinite) dose and duration of application;

- temperature and humidity conditions;

- sample collection and analysis;

- presentation of the results and content of the final report.]

To support this guideline COLIPA submitted to the OECD, ECVAM and
the SCC a dossier consisting of "in-house" data and articles from the
literature designed to demonstrate the predictive value, reliability and
robustness of the in vitro approach.

It may be summarised as follows:

- Predictive value

The in vitro tests are useful predictors of in vivo cutaneous
absorption.

[Bearing in mind the variability of in vivo data, there is good
correlation between

     - in vitro (pig) and in vivo (rat) data for 10+8 hair dyes;

     - in vitro (rat) and in vivo (rat) data for 8 coded substances of
different solubility;

     - in vivo (pig) and in vitro (man) data for 7 colouring agents;

     - in vitro (pig) and in vivo (man) data for 1 UV filter in
ethanol solution.

The skin of the pig's ear is a very good model of the human skin in in vitro
cutaneous absorption tests of oestradiol.]

It is possible to design prediction models of skin pharmacokinetics on the
basis of in vitro tests.

[The in vitro data in the rat can be used in combination with a skin
pharmacokinetic mode.

The model correctly predicts experimental in vivo measurements in man
for an ester in suspension in a mixture of organic solvents and surfactants.]

- _Reliability_

"In-house" experience in this domain pertained to different
categories of cosmetic ingredients, including UV filters, hair dyes,
surfactants, mineral hydrocarbons, etc.

- _Robustness_

**28,**

The dossier contains scanty information on the intra- and
interlaboratory variability of the results and their statistical
analysis.

[The skin penetration coefficients (Kp) and their standard
deviations give an idea of the degree of reproducibility of the
results obtained in a series of in vitro tests conducted on benzyl
alcohol. Good interlaboratory reproducibility is observed in _one_
study conducted in two laboratories using the same in vitro
protocol (rat) to measure the skin penetration of two UV filters.]

- Influence of operating conditions

There are some articles in the literature that provide patchy and
unsystematic information on the influence of storage conditions,
the type of cell used, and the batches of skin used.

[- The use of the excised skin of fresh or deep-frozen pig does
not influence the results for two UV filters (lipophiles) and
lactic acid (a hydrophile);

     - the type of cell used (static or continuous flux) does not
influence the cutaneous absorption of water-soluble
substances through various types of membranes taken from
different species (pig, rat, man) into different destination
solutions;

     - the choice of the destination solution is crucial for

substances which have low solubility in water;

     - batches of skin taken from three different animals (rats) do
not influence the data on cutaneous absorption and
distribution through the skin in respect of two UV filters
tested using the same vehicle.]

_OECD_

The USA and the EU are at odds about the in vitro approach within the
OECD's group of national coordinators.

**At its 7th meeting,** held in Paris on 18 and 19 September 1996, **the group**
**of national coordinators decided:**

**1.** **to devote a forthcoming workshop to the study of existing in**
**vitro percutaneous absorption data;**
**2.** **to base the decisions on** a guideline document to be prepared in
the coming year by the OECD Secretariat on **the criteria for the**
**validation and acceptance of alternative methods** (on the basis
of the Solna workshop report);
3. to develop a dual guideline on in vivo and in vitro tests, in the
event of rapid development of the in vitro dossier.

_SCC_

**29**

**Although** the SCC has **emphasised** that **the test protocols used by**
**industry were not subjected to a formal validation test** and although it
recommended that the existing documentation be supplemented, notably
as regards intra- and intra-laboratory reproducibility and the influence of
the vehicle on the release of the cosmetic ingredients, **the SCC is**
**convinced of the relevance of the in vitro methods** and has in recent

years agreed to draw on in vitro percutaneous absorption data in the
evaluation of the safety of several cosmetic ingredients for human use.

3. **Eye irritation**

_ECVAM/COLIPA_

**The international EC/HO validation study of alternatives to the**
**Draize eye irritancy test did not achieve the expected objectives but it**
**inspired the organisation of an** **ECVAM/ERGATT** **workshop on the**
**practical aspects of validation and the preparation of a prevalidation**
**schedule by ECVAM,** as well as the planning of the COLIPA study.

**Limited results were obtained** for a small number of protocols **in**
**COLIPA's first international validation phase** of alternatives to the
Draize test, where prediction models had been prepared for each test.

Statistical evaluation of the results of the first validation phase indicate
that the most promising protocols are FLtest [2], TEA [3] and RBCtest, [4] which
conform reasonably with their prediction models; however, for the first
two more reproducibility data is required because in each case they have
only been used by two large laboratories, while the utility of the FL and
RBC tests has been challenged because the pertinence of their
classification models is not universally supported. Moreover, no attempt
has been made to evaluate the idea of a battery of two or more tests to
replace in vivo assays.

**Currently there are no validated alternative methods capable of**
**replacing the** **OECD-405** **in vivo eye irritancy test.**

**4.** **Skin irritation**

_ECVAM_

Fluorescein leakage test, measures the damage caused to a cell barrier.

Tissue Equivalent Assay, measures the time required to cause a 50% reduction in the viability of
cells in reconstituted human skill.

Red blood cell haemalysis test, used to evaluate damage to the cell membranes.

50.

Following a prevalidation exercise in 1995 on skin corrosion tests (TER,
CORROSITEX and SKIN and EPISKIN), **ECVAM undertook a formal**
**validation study** involving four skin corrosion tests and 60 test materials,
whose results are expected in 1997.

Considerable progress has been made in developing a database to predict
irritation potential. However the in vitro tests currently being validated
concern highly irritant substances and so this research - which is very
useful for classifying and labelling dangerous substances - is only of
limited use in evaluating the skin tolerance of cosmetic ingredients, which
are a priori weakly- or non-irritant substances (at their concentration of
use).

**There are to date no validated alternative methodologies capable of**
**replacing the OECD 404 in vivo skin irritancy test. An** **ECVAM**
**workshop on human skin equivalents and their uses is planned for**
**1997.**

_OECD_

A **draft test strategy** for determining the skin irritation potential of
chemical substances is **on the programme of the OECD** group of
national coordinators.

_The strategy proposed by the OECD envisages first eliminating the most_
_irritant substances through examination of the structure activity relations_
_and structure property relations, as_ _well_ _as by using information derived_
_from in vitro corrosion tests, before considering recourse to conventional_
_tests on rabbits, in_ _vivo,_ _or the use of human volunteers._

5. **Skin sensitisation**

Identifying skin irritation potential which manifests itself as contact
allergy is important in evaluating the safety of cosmetic ingredients.

The impressive research work done up to now has helped clarify our
understanding of the mechanistic bases of skin sensitisation. It seems that
predicting sensitisation potential is feasible only by combining
information derived from several supplementary tests.

_ECVAM_

The report of ECVAM workshop 19 reviews the situation and contains
recommendations for research aimed at replacing current animal tests,
viz.:

            - refine the DEREK expert system identifying the
structural alerts of skin sensitisation and develop
supplementary systems based on the SARs
(Structure Activity Relationships)

**31**

             - standardise the tests "on cultures" of the most

promising in vitro cells, and notably study the
essential role of mediators derived from human

dendritic cells and standardise the methods of

isolating and cultivating these cells

             - develop other in vitro models, notably on skin
expiants

             - develop an in vitro skin metabolism system

             - develop an adequate database on the basis of these
tests, through a validation exercise using a suitable
in vivo database.

ECVAM hopes to support a prevalidation study on an in vitro test, in
1997.

Bearing in mind the complexity of the process and the fact that predicting
sensitising potential is a difficult task because of the variability of
individual response, **definitive results in this domain can** be **expected**
**only in the medium** **term,** **if not indeed the long term.**

**6.** **Skin compatibility of the finished product**

The guidelines prepared by COLIPA in November 1995 on evaluating the
skin compatibility _oi finished cosmetic products_ in the framework of
strictly controlled clinical studies, which were informed by ethical
considerations and based on prior knowledge of the composition and
stability of the tested products and prior evaluation of the toxicity data
pertaining to the ingredients used in the product, have been published in
_Food and Chemical Toxicology_ [34 (1996) 651-660].

#### ISSN 0254-1475

## COM(97) 182 final

# DOCUMENTS

### EN 10 15 02 Catalogue number : CB-CO-97-176-EN-C ISBN 92-78-19036-5

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