National Toxicology Program

National Toxicology Program

Regulatory Acceptance of Alternative Methods

http://ntp.niehs.nih.gov/go/regaccept

ICCVAM was established to facilitate the review and adoption of scientifically valid safety testing methods designed to protect human health, animal health and the environment while refining, reducing or replacing animal use where feasible. Below is a list of alternative safety testing methods that are accepted by U.S. and international regulatory authorities. Appropriate use of these test methods can significantly reduce animal use and improve animal welfare. ICCVAM has also identified critical research, development, and validation efforts needed to further advance numerous other alternative methods.

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Acute Dermal Systemic Toxicity

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement

OECD Adoption

EU Regulatory Acceptance/ Endorsement

In vitro dermal absorption methods

ICCVAM working group contributed to U.S. OECD test guideline review, expert consultation meetings

Accepted by U.S. via OECD TG 428

OECD TG 428 (2004)

Via OECD

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Acute Oral Systemic Toxicity

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Up-and-down procedure for acute oral toxicity

ICCVAM peer review and report; recommended in 2001

Accepted by U.S. in 2003; EPA OPPTS 870.1100 (2002)

OECD TG 425 (2001)

Via OECD

Fixed dose procedure for acute oral toxicity

ICCVAM working group contributed to test guideline development

Accepted by U.S. via OECD TG 420

OECD TG 420 (2001)

Via OECD

Acute toxic class method for acute oral toxicity

ICCVAM working group contributed to test guideline development

Accepted by U.S. via OECD TG 423

OECD TG 423 (2001)

Via OECD

Acute toxicity in vitro starting dose procedure, 3T3 cells

ICCVAM 2001 workshop report; ICCVAM 2006 peer review and report; recommended in 2008

Accepted by U.S. agencies in 2008

OECD GD 129 (2010)

Via OECD

Acute toxicity in vitro starting dose procedure, NHK cells

ICCVAM 2001 workshop report; ICCVAM 2006 peer review and report; recommended in 2008

Accepted by U.S. agencies in 2008

OECD GD 129 (2010)

Via OECD

Avian acute oral toxicity test (reduction of animal use)

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 223

OECD TG 223 (2010)

Via OECD

Harmonized guidance for nonclinical safety studies for pharmaceuticals (reduction of animal use)

FDA activity

Accepted by FDA via ICH in 2010

ICH Guidance Document M3(R2)

 

Waiving or bridging of mammalian acute toxicity tests for pesticides and pesticide products (reduction of animal use)

EPA activity

EPA guidance (2012) provides for authorizing exemptions from standard requirements under 40 CFR 158.500, 40 CFR 161.340, other regulations and notices

NA

 

In vitro post-column oxidation test method for paralytic shellfish toxin detection

ICATM activity

(may be used on an interim basis while awaiting implementation)

Association of Analytical Communities official method (2011); Canadian Food Inspection Agency (2011)

 

Harmonized guidance for preclinical safety evaluation of biotechnology-derived pharmaceuticals (reduction of animal use)

FDA activity

Accepted by FDA via ICH in 2012

ICH Guidance Document S6(R1)

 

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Acute Inhalation Toxicity

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Inhalation toxicity - acute toxic class method

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 436

OECD TG 436 (2009)

Via OECD

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Dermal Phototoxicity

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

3T3 NRU phototoxicity test for skin photo-irritation

ICCVAM contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 432

OECD TG 432 (2004)

Via OECD

3T3 NRU phototoxicity test:  application to UV filter chemicals

ICCVAM contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 432

OECD TG 432 (2004)

Via OECD

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Dermal Corrosivity and Irritation

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Corrositex® in vitro membrane barrier skin corrosivity test

ICCVAM peer review and report; recommended in 1999

Accepted by U.S. agencies in 1999; 49 CFR 173.137 (2011)

OECD TG 435 (2006)

Via OECD

EpiSkin™ in vitro human skin model skin corrosivity test

ICCVAM review and report; recommended in 2002

Accepted by U.S. via OECD TG 431; 49 CFR 173.137 (2011)

OECD TG 431 (2004)

Via OECD

EpiDerm™ in vitro human skin model skin corrosivity test

ICCVAM review and report; recommended in 2002

Accepted by U.S. via OECD TG 431; 49 CFR 173.137 (2011)

OECD TG 431 (2004)

Via OECD

SkinEthic™ in vitro human skin model skin corrosivity test

ICCVAM contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 431 (meets performance standards 2006)

OECD TG 431 (2004)

Via OECD

Rat TER in vitro skin corrosivity test

ICCVAM review and report; recommended in 2002

Accepted by U.S. via OECD TG 430

OECD TG 430 (2004)

Via OECD

EST-1000 in vitro test method for skin corrosivity testing

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 431 (meets performance standards 2009)

OECD TG 431 (2004)

Via OECD

EpiSkin™ in vitro human skin model skin irritation test

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 439

OECD TG 439 (2010)

Via OECD

EpiDerm™ in vitro human skin model skin irritation test

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 439

OECD TG 439 (2010)

Via OECD

SkinEthic™ in vitro human skin model skin irritation test

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 439

OECD TG 439 (2010)

Via OECD

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Immunotoxicity: Allergic Contact Dermatitis

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Murine local lymph node assay (LLNA) for skin sensitization

ICCVAM peer review and report; recommended in 1999

Accepted by U.S. agencies in 1999; EPA OPPTS 870.2600 (2003) and FDA Guidance for Industry: Immunotoxicology Evaluation of Investigational New Drugs (2002)

OECD TG 429 (2002) ISO (2002)

Via OECD

Updated LLNA protocol (requires 20% fewer animals)

ICCVAM peer review and report; recommended in 2009

Accepted by U.S. agencies in 2010; EPA updated policy on the use of the LLNA for end-use pesticide products in 2011

OECD TG 429 (2010)

Via OECD

Reduced LLNA protocol (requires 40% fewer animals by using only the high dose group)

ICCVAM peer review and report; recommended in 2009

Accepted by U.S. agencies in 2010; EPA adopted the reduced LLNA in 2011

OECD TG 429 (2010)

Via OECD

LLNA: DA for skin sensitization testing (a nonradioisotopic LLNA test method)

ICCVAM peer review and report; recommended in 2010

Accepted by U.S. agencies in 2010

OECD TG 442A (2010)

Via OECD

LLNA: BrdU-ELISA for skin sensitization testing (a nonradioisotopic LLNA test method)

ICCVAM peer review and report; recommended in 2010

Accepted by U.S. agencies in 2010

OECD TG 442B (2010)

Via OECD

LLNA for potency categorization of skin sensitizers (refinement and reduction of animal use)

ICCVAM peer review and report; recommendations in 2011

Accepted by U.S. agencies in 2012

GHS (2009)

Via GHS

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Ocular Corrosivity and Irritation

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Bovine corneal opacity and permeability in vitro test method to identify severe eye irritants/corrosives

ICCVAM review and report; recommended in 2007

Accepted by U.S. agencies in 2008

OECD TG 437 (2009)

Via OECD

Isolated chicken eye in vitro test method to identify severe eye irritants/corrosives

ICCVAM review and report; recommended in 2007

Accepted by U.S. agencies in 2008

OECD TG 438 (2009)

Via OECD

Cytosensor microphysiometer in vitro test method for eye safety testing

ICCVAM peer review and report; recommended in 2010

Accepted by U.S. agencies in 2011

New OECD test guideline under consideration

 

Use of anesthetics, analgesics, and humane endpoints for in vivo eye safety testing (refinement of animal use)

ICCVAM peer review and report; recommended in 2010

Accepted by U.S. agencies in 2011

Updated OECD TG 405 (2012)

Via OECD

In vitro fluorescein leakage test method for identifying ocular corrosives and severe irritants

ICCVAM working group contributed to OECD test guideline review

Accepted by U.S. agencies via OECD TG 460

OECD TG 457 (2012)

Via OECD

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Pyrogen Testing

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Human whole blood/interleukin-1β in vitro pyrogen test

ICCVAM peer review and report; recommended in 2008

Accepted by FDA in 2009; use addressed in June 2012 FDA guidance on pyrogen testing

NA

Published in European Pharmacopeia

Human whole blood/interleukin-1β in vitro pyrogen test: application of cryopreserved human whole blood

ICCVAM peer review and report; recommended in 2008

Accepted by FDA in 2009; use addressed in June 2012 FDA guidance on pyrogen testing

NA

Published in European Pharmacopeia

Human whole blood/interleukin-6 in vitro pyrogen test

ICCVAM peer review and report; recommended in 2008

Accepted by FDA in 2009; use addressed in June 2012 FDA guidance on pyrogen testing

NA

Published in European Pharmacopeia

Human peripheral blood mononuclear cell/interleukin-6 in vitro pyrogen test

ICCVAM peer review and report; recommended in 2008

Accepted by FDA in 2009; use addressed in June 2012 FDA guidance on pyrogen testing

NA

Published in European Pharmacopeia

Monocytoid cell line Mono Mac 6/interleukin-6 in vitro pyrogen test

ICCVAM peer review and report; recommended in 2008

Accepted by FDA in 2009; use addressed in June 2012 FDA guidance on pyrogen testing

NA

Published in European Pharmacopeia

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Biologics Testing

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

ELISA test for batch potency testing of human tetanus vaccines (refinement: antibody quantification)

FDA activity

21 CFR 610.10; use reviewed on a case-by-case basis

NA

Published in European Pharmacopeia (2003)

ToBI test for batch potency testing of human tetanus vaccines (refinement: antibody quantification)

FDA activity

Per 21 CFR 610.10; use reviewed on a case-by-case basis

NA

Published in European Pharmacopeia (2003)

Use of humane endpoints in animal testing of biological products

USDA activity

Addressed in 9 CFR 117.4e, Center for Veterinary Biologics (CVB) Notice No. 04-09 (2004)

NA

 

Rabies vaccine, humane endpoints

USDA activity

Addressed in 9 CFR 117.4e, CVB Notice No. 04-09 (2004)

NA

 

Relevance of the target animal safety test for batch safety testing of vaccines for veterinary use

USDA activity

9 CFR 113.4 provides for authorizing exemptions from standard requirements

NA

Published in European Pharmacopeia (2004)

ELISA test for batch potency testing of Leptospira interrogans serovar pomona (replacement: antigen quantification)

USDA activity

USDA Supplemental Assay Method (SAM) 624 (2008)

NA

 

ELISA test for batch potency testing of Leptospira interrogans serovar canicola (replacement: antigen quantification)

USDA activity

USDA SAM 625 (2008)

NA

 

ELISA test for batch potency testing of Leptospira interrogans serovar icterohaemorrhagiae (replacement: antigen quantification)

USDA activity

USDA SAM 627 (2008)

NA

 

ELISA test for batch potency testing of erysipelas vaccines
(replacement: antigen quantification)

USDA activity

USDA SAM 613 (2008)

NA

Published in European Pharmacopeia

ELISA test for batch potency testing of Leptospira kirschneri serovar grippotyphosa (replacement: antigen quantification)

USDA activity

USDA SAM 626 (2009)

NA

 

Cell-based potency assay for stability and potency of botulinum neurotoxin type A products

FDA activity

Allergan, Inc., method accepted by FDA in 2011

NA

 

USDA guidelines on master reference qualification and requalification for vaccine potency assays (reduction of animal use)

USDA activity

Addressed in 9 CFR 113.8(d)(2), Veterinary Services Memorandum 800.211 (2011)

NA

 

USDA guidelines on use of humane endpoints and methods in animal testing of biological products (refinement of animal use)

USDA activity

Addressed in 9 CFR 117.4(e), CVB Notice No. 12-12 (2012)

NA

 

Serum neutralization test for potency testing of inactivated veterinary rabies vaccines (reduction and refinement of animal use)

ICCVAM workshop in 2011

NA

NA

Published in European Pharmacopoeia Monograph 0451 (2012)

Alternative test procedure for tuberculin, PPD Bovis, intradermic (reduces animal use by 65%)

USDA activity

Applies to 9 CFR 113.409(c), described in Veterinary Services Memorandum 800.114 (2012)

NA

 

Histamine sensitization test for acellular pertussis vaccines based on temperature measurement (refinement of animal use)

Discussed at ICCVAM workshop in 2010

NA

WHO Technical Service Report 878 (update adopted 2012)

 

USDA elimination of the upper limit LD50 for rabies vaccine potency testing (reduction of animal use)

USDA activity

Modification to SAM 308, CVB Notice No. 13-10 (2013)

NA

 

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Endocrine Disruptors

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

Uterotrophic bioassay in rodents: a short-term screening test for estrogenic properties

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 440 in 2007; EPA 890.1600 (2009)

OECD TG 440 (2007)

Via OECD

Hershberger bioassay in rats: a short-term screening assay for (anti) androgenic properties

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 441; EPA OPPTS 890.1400 (2009)

OECD TG 441 (2009)

Via OECD

Stably transfected human estrogen receptor-α transcriptional activation assay for the detection of estrogenic agonist-activity of chemicals

ICCVAM working group contributed to U.S. OECD test guideline review, expert consultation meetings

Accepted by U.S. via OECD TG 455; EPA OPPTS 890.1300 (2009)

OECD TG 455 (2009); updated 2012 to include performance standards

Via OECD

In vitro H295R steroidogenesis assay

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. agencies via OECD TG 456

OECD TG 456 (2011)

Via OECD

In vitro BG1Luc ER TA agonist assay to identify substances that induce human ER activity

ICCVAM peer review and report; recommendations in 2012

Accepted by U.S. agencies in 2012

OECD TG 457 (2011)

Via OECD

In vitro BG1Luc ER TA antagonist assay to identify substances that inhibit human ER activity

ICCVAM peer review and report; recommendations in 2012

Accepted by U.S. agencies in 2012

OECD TG 457 (2011)

Via OECD

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Genetic Toxicity

Method

ICCVAM or ICCVAM Agency Contributions

U.S. Regulatory Acceptance/ Endorsement and Applicable Regulations and Guidance

OECD Adoption

EU Regulatory Acceptance/ Endorsement

In vitro mammalian cell micronucleus test

ICCVAM working group contributed to U.S. OECD test guideline review

Accepted by U.S. via OECD TG 487

OECD TG 487 (2010); included in 2011 International Conference on Harmonisation harmonized guideline for testing human pharmaceuticals

Via OECD

The NTP is administratively located at the National Institute of Environmental Health Sciences, part of the National Institutes of Health.

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