ANIMAL TESTING SAFETY END POINTS IN CHEMICAL INDUSTRY

(Precisely EU legislation on registration, evaluation & authorisation of chemicals or REACH)

SUBMITTED BY-MIHIR PUJARA

DEPT. OF PHARMACEUTICAL MANAGEMENT,NIPER,MOHALI

FLOW OF PRESENTATION

Introduction

About REACH

REACH process in detail

What are “end points”?

Chart describing overall process related to chemical safety assessment

Testing involved in REACH proposal(annex V to X)

Acute toxicity tests

REACH-needed data of all the “end points”

Reproductive & developmental toxicology testing requirements

Examples

Brief introduction of other regulatory agencies

INTRODUCTION

CHEMICAL SECTOR ANIMAL TESTING

One of the biggest industrial sector in EU

Numerous regulations

Numerous substances

Numerous stakeholderso Different authoritieso Agencies from several countrieso Many companies/competitiorso Traderso Many customer(diverse usages

& worldwide marketing)

At least 50 new chemicals in EU eachyear. Approximately 60 000 – 100 000 chemical substances are on the market

Proper regulation is mandatory

Animal tests were developed and testing demands increased along with increasing awareness and data of toxicity of chemicals in man.

Current legislation dictates that animal test data is still required to help assess potential health hazards of chemicals to humans.

The new European REACH and CLP Regulations take this further and require approval to be obtained from ECHA, the European Chemicals Agency, before any animal tests are carried out, for both substances and preparations.

Approval for new testing on animals will only be granted once all other methods to generate the necessary data have been exhausted.

REACH is a European Union regulation concerning the Registration, Evaluation, Authorisation and restriction of Chemicals. It came into force on 1st June 2007 and replaced a number of European Directives and Regulations with a single system.

To protect human health & environment from the use of chemicalsUnderstand & manage risk associated with chemicals & their useTo allow free movement of substances on EU marketTo enhance innovation in and the competitiveness of the EU chemicals industryTo promote the use of alternative methods for the assessment of the hazardous properties of substances. e.g.QSARREACH foresees data sharing between registrants to gather the required information

The regulation applies to substances manufactured in, or imported to the EU in annual quantities of 1 tonne or more per company, unless the regulation indicates otherwise.

WHAT IS REACH?

PROPOSAL FOR SAFETY DRIVERS: WHY THERE IS SAFETY TESTING?

If the substances are not registered, then the data on them will not be available & as a result a firm will no longer be able to manufacture or supply them legally.

Pre registrationApplies from 1 june 2008Chemicals currently on EU market are pre-registered between 1

june to 1 december,2008Companies who pre-register their substances can benefit from

extended registration deadlinesLate pre registration

It applies, when any company starts manufacturing or importing 1 tonne or more chemical substance per year after 1 dec,2008Late pre-registration does not apply to companies that failed to meet the pre-registration deadline of 1 December 2008 for their substances

Chemicals manufactured or imported in large volumes have to register first

NO DATA, NO MARKET

NECESSARY THINGS FOR REGISTRATION

1.A technical dossier-for substances in quantities of 1 tonne or more per year

contains information on properties, classification, guidance & uses of chemical

Information is proportional to quantity. The higher the tonnage more information on the intrinsic properties of the chemical is required

2.A chemical safety report-for substances in quantities of 10 tonnes or more per year

documents the hazards

classify the substances and make assessment as to whether the substance is persistent, bioaccumulative and toxic (PBT) or very persistent and very bio-accumulative (vPvB).

REGISTRATION

PROCESS OF REGISTRATION

DOSSIER SUBMISSION IN IUCLID 5 FORMAT

PAYMENT OF FEES

UPDATE IF ANY CHANGES IN PRODUCTION OR IMPORT

DOSSIER COMPLETENESS CHECK

In cases where a substance is manufactured or imported by more than one company, they are required to jointly submit information on the hazardous properties of the substance and its classification. If registrants agree they can also submit a joint chemical safety report.

If the Agency does not indicate otherwise within 3 weeks of the registration, the registrant may begin (for non-phase-in substances) or continue (for phase-in substances) to manufacture or import the substance

Evaluation procedure deals with examination of testing proposals & compliance check of registrations & substancesMain objective behind evaluation:-

Reliable & adequate data are producedPrevent unnecessary animal testing

OBJECTIVES

DOSSIER EVALUATION

•Agency may select any reg. dossier to check whether the proper information is available & has been adequately reported.•They can also ask for further information if required.

COMPLIANCE CHECK CHECKING OF TESTING PROPOSALS

•For chemicals manufactured or imported in a quantity of 100 tonnes or more, a testing proposal must be submitted in the reg. dossier, agency then evaluate it.

SUBSTANCE EVALUATION•If the substance is hazardous or pose a risk to human or environment, the agency will include that substance on list for “substance evaluation”.•Then registrant will be requested to provide further information. 

EVALUATION

Substances of very high concern will be gradually included in Annex XIV of the REACH Regulation. Once included, they cannot be placed on the market or used after a date to be set (the so-called “sunset date”) unless the company is granted an authorisation.

It is essential to regulate them because the effects they can have on humans and the environment are very serious and often irreversible. There is no tonnage threshold for asubstance to be subject to authorisation.

These substances are Carcinogenic, Mutagenic or toxic to Reproduction (CMR) classified in category 1 or 2, PBT or vPvB according to the criteria in Annex XIII.

The authorisation process consist of four steps:-

Step 1: Identification of substances of very high concern (by authorities)Accordance with Annex XV, member state competent authorities prepare a

dossierThe outcome of this identification process is a list of identified substances,

which are candidates for prioritisation (the “candidate list”).

Step 2: Prioritisation process (by authorities) The substances on the candidate list are then prioritised to determine which ones

should be subject to authorisationInterested parties are invited to submit comments during this process

AUTHORISATION

Finally whether the substance is subject to authorisation or not is decided & the “sunset date” by when a substance can no more be used without authorisation is decided

Step 3:Applications for authorisation (by industry)

Applicant have to provide chemical safety report, suitable alternatives or techniques to reduce overall risks & economically feasible.

Applicant can include socio-economic analysis in his application.A fee has to be paid for each application.For all applications, the Agency will provide expert opinions. The applicant

can comment on these opinions.

Step 4: Granting of authorisations (by the European Commission)

If the risk associated with the use of substance is properly controlled authorisations will be granted.

If the risk is not adequately controlled, an authorisation may still be granted if it is proven that the socio-economic benefits outweigh the risks & there is no other option available.

All authorisations will be reviewed after a certain time-limit which will be set on a case-by-case basis.

The Annex XV dossier should demonstrate that there is a risk to human health or the environment that needs to be addressed at Community level and should identify the most appropriate set of risk reduction measures.

Annex XVII of the REACH Regulation contains the list of all restricted substances, specifying which uses are restrictede.g. the ban on asbestos and restrictions on the uses of certain azo-dyes

There is no tonnage threshold for a substance to be subject to restriction

SO WHAT IS “END POINT”?

An endpoint is an observable or measurable inherent property of a chemicalsubstance. It can for example refer to a physical-chemical property like vapour pressure or to degradability or to a biological effect that a given substance has on human health or the environment. e.g. carcinogenicity, irritation, aquatic toxicity

In order to register a substance under REACH, the registrant must submit specific information about the intrinsic properties of the substance in each of the following areas:-physical/chemical properties, human toxicological information, ecotoxicological information.Data on the intrinsic properties of a substance are categorised into endpoints. For instance, “carcinogenicity” is a human toxicological endpoint.

RESTRICTION

OVERALL REACH PROCESS-SUMMARY

PRE REGISTRATION

REGISTRATION

EVALUATION

AUTHORISATION

Consortia/partner build upInform European Chemical AgencySIEF(data exchange)

Chemical safety report to ECATesting proposalsAllocation of competent authorityPay reg. fees

Dossier audit &/or evaluationAgree testing plans

Socio economic analysisRisk management checksReplacement plansTime limit authorisation

Overall process related to information requirements and chemicals safety assessment under REACH

Ref:-http://guidance.echa.europa.eu/docs/guidance document/information requirement r2 .pdf

TESTING INVOLVED IN THE REACH PROPOSAL

Annexes V to VIII of the REACH proposal include lists of tests which will be involved in the chemical safety testing programme.

ANNEX V

• Includes standard information requirement for substances manufactured or imported in quantities of 1 tonne or more• Data will generally collected with non-animal tests

ANNEX VI,VII,VIII

• Provide additional standard information requirements for substances manufactured or imported in quantities of 10,100 & 1000 tonnes or more respectively

• Mainly based on animal tests.

ANNEX X(PREVIOUSLY ANNEX V OF THE DANGEROUS SUBSTANCES DIRECTIVE(67/548/EC)

• Details the animal protocols• Does not include all in vitro and other non-animal tests currently available.

Skin irritation or skin corrosion (in vitro)Eye irritation (in vitro)Skin sensitisation (Local Lymph Node Assay)Mutagenicity (in vitro)Proposal to include an acute toxicity test (in vivo or in vitro)

Skin irritation (in vivo)Eye irritation (in vivo)Mutagenicity (in vitro)Acute toxicity (in vivo by oral route and/or by inhalation and/or dermal route)Repeated dose toxicity (28-day in vivo study, one species, male and female)

Reproductive toxicity (Screening for developmental toxicity in vivo, if there is no (Q)SAR estimates that the substance may be a developmental toxicant. A positive result in the screening should be confirmed by further in vivo tests)Toxicokinetics – if it can be derived from the available informationShort term toxicity testing on fishAdsoption/desorption screening

Annex V – all chemicals

Annex VI – above 10 tonnes

Mutagenicity in vivo – if positive results from the Annex V and VI tests28-day repeated dose toxicity or sub-chronic toxicity study (90-day, one species, rodent)Developmental toxicity study (two species)Two-generation reproductive toxicity study (in vivo, one species, male and female)Long term toxicity testing on fish (fish early-life stage toxicity test or fish short term toxicity test onembryo and sac-fry stages or fish juvenile growth test)Accumulation in one aquatic species, preferably fishFurther studies on adsoption/desorption depending on results of studies required in Annex If appropriate, further mutagenicity studies shall be proposedA long term (12 month) repeated toxicity study may be proposedTwo generation reproductive toxicity study, one species, male and female, if not provided as part ofAnnex VII requirementsA carcinogenicity study may be proposed by the applicant or requested by the competent authorityLong term or reproductive toxicity in birds

Annex VII – above 100 tonnes

Annex VIII – above 1,000 tonnes

oThe standard testing regime for all substances may be adapted for the following accepted reasons:-

1.Existing data (physical-chemical properties, animal experiments, historical human data)

o If exposure duration is comparable & if adequate & reliable documentation is provided, data consider to be equivalent to test

2.Weight of evidenceo If newly develop test methods or other independent sources indicate that

substance has not a particular dangerous property, further testing on vertebrate animals for that property shall be omitted(and reverse is also true).

3.QSARoResults obtained from valid qualitative or quantitative structure activity

relationship models ((Q)SARs) may indicate the presence or absence of a certain dangerous property.

4.In vitro methodsoIn some cases in vitro data indicate the presence or absence of certain dangerous

property. So there is no need to carry out animal experiments.e.g. VIABLITY (end point) measured by MTT test using epidermal cells.

ANNEX IX: GENERAL RULES FOR ADAPTATION OF THE STANDARD TESTING SET OUT IN ANNEXES V TO VIII

5.Grouping of substances and read-across approacho Substances whose physicochemical, toxicological and ecotoxicological

properties are likely to be similar are considered as group and their human health effects & environmental fate may be predicted by comparing with reference substance within the group.

ACUTE TOXICITY TESTS

Acute toxicity data is mainly to label and classify chemicals based on their toxicity, for application to the human situation.

Lethality has been the primary toxicological endpoint in acute toxicity tests, which have been in use for nearly 80 years but a large number of animals has been sacrificed.

For testing acute toxicity by the oral route, the LD50 method has been replaced in the OECD guidelines by other protocols which use fewer animals.

o Fixed dose procedureo Up & down procedureo Acute toxic class method

For testing acute toxicity by inhalation or by the dermal (skin) route, LD50/LC50 tests are still most predominantly used.

These tests are highly criticised due to extent of animal suffering that it causes.

All serve to provide an estimated LD50 value

Ref:-the report of CONAM/ecopa chemical policy working group, March 2007

REACH-NEEDED DATA :“PHYSICO-CHEMICAL ENDPOINTS”

TOXICOLOGICAL ENDPOINTS

Aquatic toxicity

Growth inhibition with aquatic plants (algae)

Y Y Y

Short-term toxicity on invertebrates (Daphnia)

Y Y Y

Short-term tox, fish N N Y Y

Activated sludge respiration inhibition

N Y Y

Long-term toxicity on invertebrates

N N Y

Long-term toxicity on fish N N N Y

Fish juvenile growth test orFish short-term test on embryos and sac-fry stages or Fish early-life stage (FELS)

N

Fate and behaviour in environment

Adsorption/desorption screening

Y Y

Bioaccumulation in aquatic species, preferably in fish

N N Y

Further information on adsorption/desorption

N N Y

Short-term toxicity to invertebrates

N N Y

Effects on soil micro-organisms

N N Y

Short-term toxicity to plants

N N Y

Degradation

Biotic Y Y Y

Biotic: stimulation testing on ultimate degradation on surface water

N N Y

Soil simulation testing N N Y

Sediment simulation testing N N Y

Identification of degradation products

N N Y

Abiotic (hydrolysis as function of pH

•Accordance with REACH Annex XI Section 2, testing for specific endpoints may be omitted if it is technically not possible to conduct the study as a consequence of the properties of the substance

e.g. very volatile, highly reactive or unstablesubstances (specific cases can be found in Column 2 of REACH Annexes VII-X).

•Where possible, scientifically sound approaches to the implementation of the 3Rs (reduction, refinement or replacement of animal use) which are already stipulated under the REACH Regulation, should be used

EXAMPLE 1

Ref:-the report of CONAM/ecopa chemical policy working group, March 2007

EXAMPLE 2

CLP-New European Regulation on Classification, Labelling and Packaging of chemical substances and mixtures

• It is about the hazards of chemical substances & mixtures & how to inform others about them.

NC3Rs-The National Centre for the Replacement, Refinement and Reduction of Animals in Research

• Established by the UK Government, is an independent scientific organization

ECETOC-European Center for Ecotoxicology and Toxicology Of Chemicals • Concerned with the identification of research needs and provision of scientific

rationale for the assessment of health effects and environmental impact, and thereby to justify industry's licence and freedom to operate.

EPAA-European Partnership for Alternative Approaches to Animal Testing• Accelerate the development, validation and acceptance of alternative approaches

to further the reduction, refinement and replacement (3Rs) of animal use in regulatory testing.

CEFIC-European Chemical Industry Council• Ensuring chemical safety and maintaining animal welfare are both issues of great

societal and ethical concern.

REGULATORY AGENCIES OTHER THAN REACH INFLUENCINGCHEMICAL INDUSTRY & ANIMAL TESTING

REFRENCES

1.www.hse.gov.uk2.guidance.echa.europa.eu3.www.ficam.fi4. Ppt presentation on “regulatory animal testing” by Dr. Karsten Muller, EPAA WG4, product safety chemicals5.www.nc3rs.org.uk6.The impact of REACH-the report of CONAM/ecopa chemical policy working group, March 20077.www.intertek.com/REACH8.REACH navigator9.Regulatory Toxicology and PharmacologyJournal homepage-www.elsevier.com/locate/yrtph10.Dangerous Preparations Directivehttp://www.scienceinthebox.com/en_UK/sustainability/dangerouspreparations_en.html11.A report on-Acute toxicity testing by Animal Defenders International National Anti-Vivisection Society, London.www.ad.international.org12.www.frame.org.uk13.www.specchemonline.com