Minimising Animal Testing for the Registration, Evaluation, and Authorisation of Chemical Substances (REACH) Regulation

See Also:            Webinars            Ombudsman Decision            Achievements

When the EU passed the REACH legislation in 2006, this testing programme was intended to ensure that animals would be used only as a last resort. Yet so far, PETA UK has estimated that more than 1 million animals have died in tests for REACH. Moreover, reports published by the European Chemicals Agency (ECHA) in 2011 and 2014 revealed that 288 experiments were conducted on animals without prior approval or justification, and additional tests were conducted on rodents and rabbits despite the availability of non-animal tests. It is estimated that more than 100,000 animals were used in these experiments.

To minimise new tests on animals, REACH contains a number of specific measures and general provisions designed to establish and enforce the principle that animal testing should be performed only as a last resort. For example, read-across and weight-of-evidence approaches as well as non-animal testing methods must be used whenever possible. Furthermore, all in vivo tests conducted under REACH in European member states must be carried out in accordance with the European Directive 2010/63/EU on the protection of animals used for scientific purposes. The directive stipulates that experiments must be designed to avoid distress and unnecessary pain and suffering. Moreover, experiments on animals should not be performed if the results can be obtained by another scientifically valid method.

What does this mean for registrants?
Registrants need to understand the relevant information requirements under REACH Annexes VII to XI, appreciate the practicalities and procedures related to creating and submitting the registration dossier, and take responsibility for adhering to the legal principle of using animals only as a last resort by using alternative methods whenever possible.

The guidance below on minimising testing on animals is separated into two parts: Part One outlines specific methods for minimising the use of animals for Annex VII to X endpoints, and Part Two summarises the general adaptations listed under REACH Annex XI.

In addition to the below, registrants should review the following:

PART ONE: SPECIFIC METHODS FOR MINIMISING ANIMAL USE UNDER ANNEXES VII TO X

Annex III: Waiving Requirements Based on Criteria Met by a Substance

Registrants can waive the human health and environmental data requirements of Annex VII if their substance does NOT fulfil the criteria of Annex III for chemicals of 1 to 10 tonnes per year. For example, a reduced dataset may be submitted if there is no indication of the following:

  • The substance is likely to have carcinogenic or mutagenic properties or be toxic to reproduction (CMR) (category 1A or 1B).
  • The substance is likely to be persistent, bioaccumulative, and toxic (PBT) or very persistent and very bioaccumulative (vPvB).
  • The substance with dispersive or diffuse uses would be classified as hazardous to human health and/or as an environmental hazard under the Classification, Labelling and Packaging

ECHA has published an Annex III inventory to help registrants who manufacture or import between 1 and 10 tonnes per year decide whether they may be able to register their substances with limited information. For more information, registrants can review ECHA’s news release on waiving these endpoints based on Annex III criteria.

Annex VII and VIII Endpoints

Skin Irritation and Corrosion
If testing is required, registrants should use OECD-accepted in vitro methods in an integrated approach to testing and assessment (IATA) to predict skin irritation and corrosion for most chemical classes as well as to predict non-irritancy. The May 2016 update to the REACH annexes eliminated the in vivo testing as a standard information requirement. It is now an Annex VIII, column 2, requirement and thus required only when in vitro studies are not applicable or the results of the in vitro studies are not adequate for classification and labeling. ECHA’s endpoint-specific guidance (Chapter R.7a, version 6.0) clearly indicates that in vitro methods should be used whenever possible.  

OECD test guidelines (TGs) include the following:

For more information on using in vitro methods to predict skin irritation and corrosion, view the webinar co-hosted by the PETA Science Consortium International and Chemical Watch, or download a factsheet here.

Severe Eye Damage and Eye Irritation
If testing is required, registrants should use OECD-accepted in vitro methods in an integrated testing strategy to replace animal testing for severe eye damage as well as to predict non-irritancy. The May 2016 update to the REACH annexes eliminated the in vivo study as a standard information requirement. It is now an Annex VIII, column 2, requirement and thus required only if the in vitro studies are not applicable, the results obtained from the studies are not applicable, or the results obtained from the in vitro studies are not adequate for classification or labeling. ECHA’s endpoint-specific guidance (Chapter R.7a, version 6.0) clearly indicates that in vitro methods should be used whenever possible.

Substances that are corrosive or non-irritant can be classified using a top-down or bottom-up approach as described in the European Commission Joint Research Centre report, Alternative Methods for Regulatory Toxicology – A State-of-the-Art Review.

OECD TGs include the following:

  • OECD TG 460: Fluorescein leakage test method for identifying ocular corrosives and severe irritants
  • OECD TG 437: Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants
  • OECD TG 438: Isolated chicken eye test method
  • OECD TG 491: The short time exposure in vitrotest method for identifying (a) chemicals inducing serious eye damage and (b) chemicals not requiring classification for eye irritation or serious eye damage
  • OECD TG 492: Reconstructed human cornea-like epithelium (RhCE) test method for identifying chemicals not requiring classification and labelling for eye irritation or serious eye damage
  • OECD QSAR Toolbox

In addition to the OECD TGs, the cytosensor microphysiometer method is recommended by EURL ECVAM to differentiate water-soluble ocular corrosives, severe irritants, and non-irritants in top-down and bottom-up approaches, respectively. A draft OECD TG for the cytosensor microphysiometer is available.

For more information on using in vitro methods to predict severe eye damage and eye irritation, view the webinar co-hosted by the Consortium and Chemical Watch, or download our factsheet.

Skin Sensitisation
The updated Annex VII requirements for skin sensitisation entered into force in October 2016, making non-animal testing the default requirement. The updated data requirements include in vitro methods for predicting skin sensitisation. An in vivo study will be required only if the in vitro or in chemico test methods are not applicable or if the results obtained from those studies are not adequate for classification and risk assessment. The updated REACH annex and legislation are available here.

Testing is not required if the substance is classified as corrosive (Category 1), is spontaneously flammable in air or in contact with water or moisture at room temperature, is a strong acid (pH ≤ 2.0), or is a strong base (pH ≥ 11.5).

Registrants should use the OECD adverse outcome pathway for skin sensitisation, which describes the key events in an adverse response, starting with the molecular initiating event and ending with the adverse health effects in humans.

In vitro OECD test guidelines (TG) include the following:

For more information on using in vitro methods to predict skin sensitisation, view the webinar co-hosted by the Consortium and Chemical Watch, or download a factsheet.

Mutagenicity
If in vitro testing under Annex VIII triggers in vivo mutagenicity testing, registrants must submit a testing proposal to ECHA for approval prior to conducting testing.

In vitro OECD TGs include the following:

Acute Systemic Toxicity
Acute systemic toxicity testing generally does not need to be conducted if the substance is classified as corrosive to the skin. Consult ECHA’s endpoint-specific guidance (Chapter R.7a, version 6.0) for more information on using a weight-of-evidence approach, which includes the use of existing data, exposure information, read-across, in silico methods, and/or in vitro methods, to fulfil data requirements for acute systemic toxicity.

Acute Oral Toxicity
Registrants should use the 3T3 neutral red uptake (NRU) cytotoxicity test to predict non-toxic chemicals as part of a weight-of-evidence approach and to support read-across arguments.

If 28-day oral toxicity test data exist or registrants are required to conduct this testing, registrants should use the results in a weight-of-evidence approach for waiving acute toxicity when the substance is anticipated to be non-toxic (see Gissi et al., 2016).

If an in vivo test is perceived to be required by regulators, the results of the 3T3 NRU test should be used to set starting doses in an in vivo study.

For more information, view the acute systemic toxicity webinar hosted by the Consortium and Chemical Watch, and see here for information about a Consortium co-sponsored workshop and other activities related to alternative approaches to acute systemic toxicity testing.

Acute Dermal Toxicity
Substances that do not meet the criteria for classification as acutely toxic or specific target organ toxicity, single exposure (STOT SE) by the oral route do not require testing for acute dermal toxicity as a second route of administration. The updated annex was published in May 2016.

Dermal toxicity can be waived based on low dermal penetration (OECD TG 428 and OECD GD 28).

Acute Inhalation Toxicity
Consider waiving acute inhalation studies based on substance vapour pressure and whether aerosols, particles, or droplets are an inhalable size. If inhalation toxicity testing is required by regulators, use the acute toxic class method OECD TG 436 instead of OECD TG 403.

Registrants should also use the OECD QSAR Toolbox to fill in any data gaps.

See here for a series of Consortium co-sponsored webinars describing alternative approaches to acute inhalation toxicity for the purpose of addressing global regulatory and non-regulatory data requirements.

Repeated Dose and Reproductive Toxicity
Screening for reproductive/developmental toxicity testing (OECD TG 421OECD TG 422) does not need to be conducted if (1) the substance is known to be a genotoxic carcinogen or germ cell mutagen and appropriate risk management measures are implemented; (2) relevant human exposure can be excluded in accordance with Annex XI, section 3; (3) a pre-natal developmental toxicity study is available; or (4) either a one-generation reproductive toxicity study or a two-generation study is available.

If screening for reproductive/developmental toxicity is required but repeated dose data is available, registrants should conduct the OECD TG 421 reproduction/developmental toxicity screening test.

If both repeated dose toxicity and reproductive toxicity studies need to be conducted, registrants should conduct the “combined” study OECD TG 422 in place of OECD TG 421 to reduce the number of animals used.

ECHA’s endpoint-specific guidance (Chapter R.7a, version 6.0) was updated in July 2017.

Short-Term Aquatic Toxicity
Testing does not need to be conducted if the substance is highly insoluble in water or unlikely to cross biological barriers or if a long-term fish toxicity study is already available.

If testing is required, use the fish embryo toxicity test (OECD TG 236 and EURL ECVAM recommendation) to predict fish acute toxicity as an alternative to the fish acute toxicity test (OECD TG 203).

If it is not possible to use the fish embryo test, the threshold approach for acute fish toxicity testing should be used.

Registrants should use the OECD QSAR Toolbox to fill in data gaps. More information on QSAR tools is available here.

A webinar co-hosted by the Consortium and Chemical Watch and detailing the current requirements and validated non-animal methods for short-term aquatic toxicity is available here.

Annex IX and X Endpoints

For substances manufactured or imported at 100 tonnes per annum or more, testing proposals must be submitted to ECHA and permission granted before testing can take place.

Repeated Dose and Reproductive Toxicity
The reproductive/developmental toxicity study does not need to be conducted if the substance is known to be a genotoxic carcinogen or germ cell mutagen and appropriate risk management measures are implemented or, as specified under Article 13 of REACH, if relevant human exposure can be excluded in accordance with Annex XI, section 3.

Furthermore, the study does not need to be conducted if the substance is of low toxicological activity (no evidence of toxicity is seen in any of the tests available), it can be proved from toxicokinetic data that no systemic absorption occurs via relevant routes of exposure, or there is no significant human exposure.

If both repeated dose toxicity and reproductive toxicity studies must be conducted for substances at 100 tonnes per annum or more (Annexes IX and X), the existing guidance for reducing the number of animals used for these endpoints is to propose long-term repeated dose toxicity studies and standard reproductive and developmental toxicity studies and not shorter-term studies, as described in Annexes VII and VIII.

Currently, the standard test required under Annexes IX and X is the OECD TG 443: the Extended One-Generation Reproductive Toxicity Study (EOGRTS). ECHA’s endpoint-specific guidance (Chapter R.7a, version 5.0) was updated in July 2017.

The requirement for a reproductive toxicity study (Section 8.7.3 of Annex IX) may be waived for substances registered under Annex IX (ie, between 100 and 1000 tonnes per annum) if no adverse effects on the reproductive tissues are observed in the available repeated dose studies (28-day or 90-day) or screening studies for reproductive effects. The reproductive toxicity study would only need to be conducted if the tonnage reached over 1000 tonnes per annum.

Long-Term Aquatic Toxicity
Testing does not need to be proposed unless required to clarify risks.

If testing is required, registrants should consider EURL ECVAM recommendations for waiving long-term aquatic toxicity studies.

Registrants should use the OECD QSAR Toolbox to fill in data gaps.

Bioaccumulation in Fish
Testing need not be proposed if (1) the substance has a low potential for bioaccumulation (eg, a log Kow < 3), (2) the substance has a low potential to cross biological membranes, or (3) direct and indirect exposure of the aquatic compartment is unlikely.

If testing is required, registrants should use the revised OECD TG 305 to reduce the number of fish used.

Registrants should also consider using information on absorption, distribution, metabolism, and excretion processes ascertained through in vitro methods to improve the prediction of bioconcentration factor models. The OECD Working Group of the National Coordinators of the Test Guidelines Programme approved a project on the development of a new OECD test guideline on in vitro fish hepatic metabolism.

Registrants should use the OECD QSAR Toolbox to fill in data gaps.

Long-Term Toxicity to Birds
Testing need not be proposed if risks to birds can be assessed using existing mammalian toxicity data.

PART TWO: GENERAL OPPORTUNITIES FOR MINIMISING ANIMAL USE UNDER ANNEX XI

In addition to the specific methods described in Part One, Annex XI outlines general procedures that registrants may use to waive the standard testing regimen and thus minimise testing on animals. These procedures can be applied individually or, in a weight-of-evidence argument, in lieu of new animal testing.

Use of Existing Data
To enable a decision on classification and labelling to be made and/or to conduct a chemical safety assessment (CSA), historical data may be considered if it is of sufficient quality (ie, reliable and valid).

Weight of Evidence
A number of sources when taken alone may not provide sufficient data on which to base an assumption or conclusion but when taken together may provide an effective means of avoiding new studies or testing proposals involving animals. Such an approach may be used for new testing methods that are not yet recognised in the Test Method Regulation (TMR) or that are internationally accredited as evidence that a substance possesses, or does not possess, a particular dangerous property.

Quantitative Structure Activity Relationships (QSARs)
The results from QSARs, which are considered adequate for the purpose of classification and labelling and/or risk assessment, could be used to fulfil one or more of the REACH information requirements or help build chemical categories without the need for animal tests.

A repository of acceptable QSARs for use by potential registrants is the OECD QSAR Toolbox, developed by the OECD to incorporate information and tools from various sources into one program for ease of use.

For more information on QSARs download a factsheet here.

Grouping and Read-Across
To avoid testing every substance for every endpoint, data gaps can be filled in by reading across from existing data for substances that are grouped together in a chemical category because they share structural similarities or common metabolic pathways. However, with ECHAꞌs narrow interpretation of read-across, it is essential that registrants provide a scientifically robust justification for waiving new animal tests and refer to ECHA’s Read-Across Assessment Framework.

Registrants should use OECD No 194: Guidance on Grouping of Chemicals, Second Edition.

In Vitro Methods
For positive results, an in vitro test is considered acceptable if it meets the EURL ECVAM criteria for acceptance into the pre-validation process. For negative results to be accepted, the test must have been scientifically validated (eg, methods with EURL ECVAM recommendations), the results should be adequate for classification and labelling and/or use in a CSA, and adequate documentation must be available. In vitro tests that have been successfully validated must be used in preference to the equivalent in vivo test.

Testing Is Not Technically Possible
Some tests may be excluded or adapted if conducting the test is not technically possible or the substance is very hazardous (eg, because it could generate an explosive atmosphere). The TMR or OECD test guidelines should be consulted, as some test methods, such as the acute dermal and acute inhalation tests, identify specific circumstances under which testing is inappropriate.

Exposure-Based Waiving
Exposure-based waiving of certain tests may be permitted in cases where it can be shown that exposure to the substance is insignificant or absent in all scenarios of its manufacture and all identified uses. This waiving is particularly relevant for preventing multiple routes of administration in acute systemic toxicity testing.

KEY MESSAGES FOR REGISTRANTS

  • Before concluding that there is an information gap for toxicity, the Substance Information Exchange Forum (SIEF) should consider all the available existing information.
  • If the substance being registered is a non-phase-in substance, the registrant should contact all other registrants listed in the response from ECHA to the Article 26 inquiry submitted by the registrant to ensure that animal testing has not already been initiated to fill in data gaps.
  • Registrants should consider whether animal testing can be minimised by following one or more of the specific testing adaptations provided in Part One above.
  • Registrants should consider whether animal testing can be omitted in more general ways as described in Part Two above.
  • If adaptation is not possible and a new animal test is perceived to be required, registrants should ensure that the least severe test using the fewest animals is employed. The test should be one that is expected to cause the least pain, suffering, distress, and lasting harm.
  • If testing is perceived to be required, registrants should check the OECD website for the most updated versions of all documents.

Further information on in vitro and in silico testing methods, including links to contract research organisations offering such testing services, is available here and here.

 

18 August 2020

Cosmetics ingredients to be tested on animals under REACH

In August 2020, two decisions published by the European Chemicals Agency (ECHA) Board of Appeal ruled that ingredients used solely in cosmetics can be tested on animals under the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) regulation. Tests on animals for cosmetics ingredients have been banned in the EU since 2013 under the Cosmetics Regulation, but these decisions effectively ignore the ban.

As a direct result of these decisions, more than 5,500 rats, rabbits, and fish are required to be used in new tests with two cosmetics ingredients: 2-ethylhexyl salicylate and homosalate. The decisions also open the door to more cosmetics tests on animals under REACH.

The PETA International Science Consortium Ltd. intervened in the ECHA Board of Appeal cases concerning these testing decisions. Although the Board of Appeal rejected many of the arguments put forward by the Science Consortium and the company responsible for appealing the testing decisions, the Science Consortium and its members are exploring options to resolve the issue.