Table of Validated and Accepted Alternative Methods

Validation & Regulatory Acceptance Status of Alternative Test Methods & Testing Strategies

(Last Updated: February 2013)

 ¹ In vitro and ex vivo methods, and in vivo methods proposed to reduce or refine animal use, are listed
 ² National and regional regulatory acceptance is not fully documented here
 ³ Replaces animal use for initial dose setting, but in vivo test required to complete assessment
 ⁴ Three cell-based CTA assays can be used to partially replace or reduce animal use
 ⁵ Screening assays to be used as part of a broader testing strategy
 ⁶ BG1Luc ER TA test method has been adapted to a high throughput screening (HTS) format and it is expected to be incorporated into Tox21 screening
 ⁷ Recommended for use as initial step within a Top-Down Approach to identify ocular corrosives and severe irritants (EU R41, GHS Category 1, and EPA Category I) for water-soluble chemicals and/or as initial step within a Bottom-Up Approach to identify non-irritants (EU:NC; GHS: NC; EPA: cat IV) for water-soluble surfactants and water-soluble surfactant-containing mixtures; does NOT correctly identify moderate and mild ocular irritants (EU: R36; GHS: Cat 2A/B; EPA: Cat II/III) so can be used for only two of the three EU and GHS classification categories for ocular irritation; cannot be used for default categorization; additional limitations on equipment availability
 ⁸ Can be used as screening test to distinguish water-soluble surfactant chemicals and certain types of surfactant-containing formulations that are not labeled as irritants (i.e., EPA Category IV, EU Not Labeled, FHSA Not Labeled) from all other hazard categories (i.e., EPA Category I, II, III; EU R41, R36; FHSA Irritant) for hazard classification and labeling under EPA, EU and FHSA classification systems; high false negative rate (24%-40%) for non-surfactant substances and formulations; high false positive rate (50% to 69%) for substances not labeled as irritants. Can be used as a screening test to identify water soluble substances as ocular corrosives and severe irritants (i.e., EPA Category I, EU R41, GHS Category 1) in tiered-testing strategy as part of weight-of-evidence approach; negative results need to be tested in another test method
 ⁹ Recommended for use as initial step within a Top-Down Approach to identify ocular corrosives and severe irritants (EU R41, GSH Category 1, and EPA Category I) for water-soluble chemicals; further refinement with respect to variability and applicability domain recommended
 ¹⁰ Restricted to use of existing data only; retrospective LVET data can be used in weight-of-evidence approach to identify potential ocular irritants
 ¹¹ rLLNA can be used for hazard classification when dose-response information is not needed
 ¹² Subject to product-specific validation to demonstrate equivalence to the rabbit pyrogen test (RPT)
 ¹³ Statement on 2 reference chemicals for in vitro skin corrosion testing
 ¹⁴ Recommended as screening test or as part of sequential testing strategy; only positive test results accepted in the 2007 endorsement
 ¹⁵ Cell-based assay for botulinum toxin potency testing is limited to products produced using the Allergan method
 ¹⁶ Contributed to OECD TG development and/or review
 ¹⁷ Draft OECD GD 43, see page 18

Supplemental Information

A. ECVAM: Summary of test methods endorsed by ECVAM and accepted by EU regulatory authorities; ESAC statements; DB-ALM (DataBase service on ALternative Methods); TSAR (Tracking System for Alternative test methods Review)

B. ICCVAM: Summary of test methods evaluated by ICCVAM and accepted by US regulatory authorities; table showing validation and acceptance status of all methods reviewed; ICCVAM-recommended protocols

C. JaCVAM: Summary of test methods evaluated and accepted by Japan's regulatory authorities

D. OECD Test Guidelines (TGs), Guidance Documents, Draft Test Guidelines

E. ICH test guidelines: technical requirements for medicinal products containing new drugs accepted by the regulatory bodies of EU, Japan, and USA

F. VICH test guidelines: EU-Japan-USA programme aimed at harmonising technical requirements for veterinary product registration

G. ISO: toxicity test guidelines for the biological evaluation of medical devices and ecotoxicity (soil quality, water quality, fire effluents, biocides used for anti-fouling systems on ships)

Acronyms

CFR: Code of Federal Regulations
EMA: European Medicines Agency
EPA: US Environmental Protection Agency
ESAC: ECVAM Scientific Advisory Committee
EURL ECVAM: European Union Reference Laboratory for alternatives to animal testing
FDA: United States Food and Drug Administration
GD: OECD Guidance Document
GHS: Globally Harmonized System of Classification and Labeling of Chemicals
ICCVAM: US Interagency Coordinating Committee on the Validation of Alternative Methods
ICH: International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use
ISO: International Organization for Standardization
JaCVAM: Japanese Center for the Validation of Alternative Methods
OECD: Organisation for Economic Cooperation and Development
OPPTS: US EPA's Office of Prevention, Pesticides and Toxic Substances
PMDA: Japan’s Pharmaceuticals and Medical Devices Agency
TG: OECD Test Guideline
USDA: United States Department of Agriculture
VICH: International Cooperation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products