Alternative Methods Validated For Regulatory Use

Alternative methods endorsed by the Organisation for Economic Co-operation and Development (OECD), the European Union Reference Laboratory for alternatives to animal testing (EURL ECVAM), the Japanese Center for the Validation of Alternative Methods (JaCVAM), the Korean Center for the Validation of Alternative Methods (KoCVAM), and/or the U.S. Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) are listed below.

See here for an 11×17 printable poster describing non-animal test strategies for eye irritation, skin irritation, and skin sensitisation.

Download an A4 or 8.5×11 printable version of the table below and skin and eye irritation and corrosion factsheets.

The table below is available in these languages.
To read in English, click on the format option: A4 or 8.5×11.
Zum Lesen auf Deutsch, hier klicken.
Para ler em português, clique aqui.
Para leer en español, haga clic aquí.
要用繁體中文閱讀,請點擊這裡

See also individual factsheets on:
EYE IRRITATION (here)
SKIN IRRITATION (here, training tool also available here)
SKIN SENSITISATION (here)
PYROGENICITY (here)
QSARS, GROUPING & READ-ACROSS (here)
EPA OPP EYE HAZARD (here)
MEDICAL DEVICES (here)
3D TISSUE MODELS (here)
ORGANS-ON-CHIPS (here)
GENOTOXICITY (here)

The following factsheets are available in French:
Les fiches de renseignements suivantes sont disponibles en français:
IRRITATION OCULAIRE (ici)
IRRITATION DE LA PEAU (ici)
SENSIBILISATION DE LA PEAU (ici)
PYROGÉNICITÉ (ici)

TOXICITY ENDPOINTTEST METHODS AND APPROACHESRECOMMENDATIONS AND STANDARD METHODS
OECDOTHER AUTHORITY
SERIOUS EYE DAMAGE AND IRRITATIONIntegrated approach on testing and assessment (IATA) for serious eye damage and irritationOECD guidance document (GD) 263, published in 2019_
Chemical toxicity assessment strategy_European Chemicals Agency guidance Chapter R.7a., R.7.2.8–R.7.2.11 (2017)
Use of a testing framework employing cytosensor microphysiometer (CM), BCOP, and the EpiOcular™ model for classification of pesticide products _US Environmental Protection Agency policy (2015)
Reconstructed human cornea-like epithelium (RhCE) test methodEpiOcular™ (MatTek, US)OECD test guideline (TG) 492, revised in 2019ESAC statement (2014); JaCVAM statements (2017 and 2018); KoCVAM guideline (2016)
SkinEthic™ (L’Oréal, France)
LabCyte (J-TEC, Japan)
MCTT HCE™ (Biosolution, South Korea)
Fluorescein leakage (FL) test methodOECD TG 460, revised in 2017ESAC statement (2009); JaCVAM statement (2013)
Short time exposure (STE) in vitro test methodOECD TG 491, revised in 2020ICCVAM report (2013); JaCVAM statement (2016); KoCVAM guideline (2017)
Vitrigel-eye irritancy test (EIT) methodOECD TG 494, revised in 2021-
In vitro macromolecular test methodOECD TG 496, published in 2019-
Bovine corneal opacity and permeability (BCOP) test methodOECD TG 437, revised in 2020ICCVAM report (2006); ESAC statement (2007); JaCVAM statements (2009 and 2014); KoCVAM guideline (2011)
Isolated chicken eye (ICE) test methodOECD TG 438, revised in 2018ICCVAM report (2006); ESAC statement (2007); JaCVAM statement (2009)
Cytosensor microphysiometer (CM) assay-ESAC statement (2009); ICCVAM report (2010)
SKIN CORROSION AND IRRITATIONIntegrated approach on testing and assessment (IATA) for skin corrosion and irritation OECD GD 203, published in 2014-
Chemical toxicity assessment strategy for skin corrosion and irritation-European Chemicals Agency guidance Chapter R.7a., R.7.2 (2017)
In vitro membrane barrier test Corrositex for skin corrosionOECD TG 435, revised in 2015ICCVAM report (1999); ESAC statement (2000); JaCVAM statement (2017)
In vitro skin corrosion: Reconstructed human epidermis (RhE) test EpiSkin™ (L’Oréal, France)OECD TG 431, revised in 2019ICCVAM report (2002); ESAC statement (1998); JaCVAM statement (2017)
EpiDerm™ (MatTek, US)ICCVAM report (2002); ESAC statement (2000); JaCVAM statement (2017)
SkinEthic™ (L’Oréal, France)ESAC statement (2006); JaCVAM statement (2017)
epiCS® (Phenion, Germany)ESAC statement (2009); JaCVAM statement (2017)
LabCyte EPI-MODEL24 SCT (J-TEC, Japan)-
Vitrolife-Skin™-JaCVAM statement (2008)
In vitro skin irritation: Reconstructed human epidermis (RhE) testEpiSkin™ (L’Oréal, France)OECD TG 439, revised in 2021ESAC statement (2007); JaCVAM statement (2010); KoCVAM guideline (2014)
EpiDerm™ (MatTek, US)ESAC statement (2008); JaCVAM statement (2013); KoCVAM guideline (2017)
SkinEthic™ (L’Oréal, France)ESAC statement (2008); JaCVAM statement (2013); KoCVAM guideline (2017)
LabCyte EPI-MODEL24 SIT (J-TEC, Japan)JaCVAM statement (2013); KoCVAM guideline (2017)
Skin+® (Sterlab, France)-
epiCS® (Phenion, Germany)
KeraSkin™ (Biosolution, South Korea)
SKIN SENSITISATIONAdverse outcome pathway (AOP) for skin sensitisationOECD GD 168 (Part 1, Part 2), published in 2012-
Guideline on defined approaches on skin sensitisationOECD Guideline 497, published in 2021-
Guidance on reporting of defined approaches and individual information sources to be used within
integrated approaches to testing and assessment (IATA)
OECD GD 256 (Annex I, Annex II), published in 2016-
Use of alternative approaches for skin sensitisation as a replacement for animal testing-US Environmental Protection Agency policy (2018)
Chemical toxicity assessment strategy -European Chemicals Agency guidance Chapter R.7a., R.7.3.4–R.7.3.7 (2017)
OECD QSAR ToolboxImplementing AOP workflow for skin sensitisationOECD training manual, released in 2017-
Example for predicting skin sensitisation of a mixture
Example of how to predict the skin sensitisation potential of a
chemical by read-across based on an analogue approach

In chemico assays addressing the adverse outcome pathway (AOP) key event on covalent binding to proteins Direct peptide reactivity assay (DPRA)OECD TG 442C, revised in 2021EURL ECVAM recommendation (2013); JaCVAM statement (2015); KoCVAM guideline (2016)
Kinetic DPRA
Amino acid derivative reactivity assay (ADRA)
ARE-Nrf2 luciferase test method KeratinoSens™OECD TG 442D, revised in 2018EURL ECVAM recommendation (2014); JaCVAM statement (2015); KoCVAM guideline (2017)
LuSens
In vitro assays addressing the adverse outcome pathway (AOP) key event on activation of dendritic cells Human cell line activation test (h-CLAT)OECD TG 442E, revised in 2018EURL ECVAM recommendation (2015); JaCVAM statement (2017); KoCVAM guideline (2017)
IL-8 Luc assay-
U937 skin sensitization test (U-SENS™)
PHOTOTOXICITY3T3 neutral red uptake (NRU) phototoxicity testOECD TG 432, revised in 2019ESAC statement (1997); ICH S10; KoCVAM guideline (2007)
Reactive oxygen species (ROS) assayOECD TG 495, published in 2019JaCVAM statement (2016); ICH S10
Reconstructed human epidermis phototoxicity test methodOECD TG 498, published in 2021ICH S10
SKIN ABSORPTION/PENETRATIONIn vitro diffusion methodOECD TG 428, published in 2004JaCVAM statement (2014); KoCVAM guideline (2009)
ACUTE SYSTEMIC TOXICITYGuidance for waiving tests for pesticide formulations-Canada Pest Management Regulatory Agency guidance (2013); US Environmental Protection Agency guidance for acute dermal toxicity tests (2016)
Strategy to replace, reduce, and refine the use of animals in the assessment of acute mammalian systemic toxicity-EURL ECVAM guidance (2014)
Collaborative Acute Toxicity Modeling Suite (CATMoS)-NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) (2019)
3T3 neutral red uptake (NRU) cytotoxicity test to identify substances not requiring classification-EURL ECVAM recommendation (2013)
GENOTOXICITY/MUTAGENICITYOECD QSAR Toolbox: Example for predicting Ames mutagenicity using read-acrossOECD training manual, released in 2017-
In vitro micronucleus testOECD TG 487, revised in 2016ESAC statement (2006); ICH S2(R1)
Bacterial reverse mutation testOECD TG 471, revised in 2020ICH S2(R1)
In vitro mammalian chromosome aberration testOECD TG 473, revised in 2016ICH S2(R1)
In vitro mammalian cell gene mutation testOECD TG 476, revised in 2016-
In vitro mammalian cell gene mutation tests using the thymidine kinase geneOECD TG 490, revised in 2016ICH S2(R1)
CARCINOGENICITYIn vitro cell transformation assays (CTA)OECD GD 214, published in 2015; OECD GD 231, published in 2016EURL ECVAM recommendations (2012 and 2013)
PYROGENICITYIn vitro monocyte activation tests (MAT)-ICCVAM report (2008); ESAC statement (2006); European Pharmacopoeia general chapter 2.6.30; US Food and Drug Administration guidance (2012)
HAEMATOTOXICITYCFU-GM assay-ESAC statement (2006)
REPRODUCTIVE TOXICITYEmbryonic stem cell test (EST)-ESAC statement (2001)
Micromass embryotoxicity assay (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.)
Whole rat embryotoxicity assay (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.)
ENDOCRINE DISRUPTOR SCREENINGStably transfected transactivation in vitro assays to detect oestrogen receptor agonists and antagonistsOECD TG 455, revised in 2021JaCVAM statement (2016)
H295R steroidogenesis assayOECD TG 456, published in 2011-
Stably transfected human androgen receptor transcriptional activation assayOECD TG 458, revised in 2020-
Human recombinant oestrogen receptor (hrER) in vitro assays to detect chemicals with ER binding affinityOECD TG 493, published in 2015-
Xenopus eleutheroembryonic thyroid assay (XETA) (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.)OECD TG 248, published in 2019-
AQUATIC TOXICITYOECD QSAR Toolbox: Example for predicting acute aquatic toxicity to fish of mixture with known componentsOECD training manual, released in 2017-
EnviroTox database to calculate threshold values-Health and Environmental Sciences Institute (2018)
Freshwater alga and cyanobacteria growth inhibition testOECD TG 201, published in 2011-
Daphnia sp acute immobilisation testOECD TG 202, published in 2004-
Fish cell line acute toxicity - the RTgill-W1 cell line assayOECD TG 249, published in 2021ISO 21115 standard (2019)
In vitro intrinsic clearance test using cryopreserved rainbow trout hepatocytes (Note: Animal primary cells are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test.)OECD GD 280, published in 2018-
In vitro intrinsic clearance test using rainbow trout liver S9 sub-cellular fraction (Note: Animal primary cells are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test.)OECD TG 319B, published in 2018; OECD GD 280, published in 2018-
Fish embryo acute toxicity test (FET) (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.)OECD TG 236, published in 2013EURL ECVAM recommendation (2014)
ALL ENDPOINTSGuidance on considerations for waiving or bridging of mammalian acute toxicity testsOECD GD 237, published in 2016-
Guidance for waiving or bridging of acute toxicity tests for pesticides-Health Canada guidance (2013)
Guidance on the reporting of defined approaches to be used within integrated approaches to testing and assessmentOECD GD 255, published in 2016-
Guidance on the validation of (Q)SAR modelsOECD GD 69, published in 2007-
OECD QSAR Toolbox: Guidance documents and training materialsOECD, revised in 2018-
QSAR Model Database-Maintained by the European Commission Joint Research Centre
Various modelling programs-For example, programs from Lhasa Limited, Leadscope, ScitoVation, and Simulations Plus
Guidance on the grouping of chemicalsOECD GD 194, published in 2014-
Read-across assessment framework-European Chemicals Agency guidance (2017)
Guidance on good in vitro method practicesOECD GD 286, published in 2018-
Guidance for describing non-guideline in vitro test methodsOECD GD 211, published in 2014-
Classification of mixtures based on the toxicity of ingredients-United Nations “Globally Harmonized System of Classification and Labelling of Chemicals” guidance (2015); US Environmental Protection Agency pilot program
ENDPOINT REPLACEMENT METHOD OR STRATEGY REGULATORY ACCEPTANCE
Biologics testing In vitro leptospirosis vaccine potency assay USDA Supplemental Assay Methods (SAM) 624, 625, 626 and 627
In vitro erysipelas vaccine potency assay USDA SAM 612 and 613
In vitro clostridial vaccine potency assay USDA draft SAM 220
In vitro tetanus toxoid potency assay USDA SAM 217
In vitro recombinant antibody production methods www.thepsci.eu/antibodies/
Veterinary target animal batch safety test (TABST) Can be waived following demonstration of compliance; USDA CVB memorandum 800.116
Revocation of general safety tests (GST)/abnormal toxicity tests (ATT) FDA amended biologics regulations to revoke GST (2015); all European Pharmacopoeia monographs revised to revoke the ATT (2017)
 

For all endpoints, in vitro methods developed in-house should always be used.

Researchers should make every effort to use available non-animal methods. If these methods are not accepted by regulatory agencies, information on reduction and refinement methods can be found here:

 

Toxicity Endpoint: Eye Irritation and Corrosion

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Integrated approach on testing and assessment (IATA) for serious eye damage and irritation OECD guidance document (GD) 263, published in 2017
Chemical toxicity assessment strategy European Chemicals Agency guidance Chapter R.7a., R.7.2.8–R.7.2.11 (2017
Use of a testing framework employing cytosensor microphysiometer (CM), BCOP, and the EpiOcular™ model for classification of pesticide products US Environmental Protection Agency policy (2015)
Reconstructed human cornea-like epithelium (RhCE) test method (EpiOcular™, SkinEthic™, LabCyte, MCTT HCE™) OECD test guideline (TG) 492, revised in 2019 ESAC statement (2014); JaCVAM statements (2017 and 2018); KoCVAM guideline (2016)
Fluorescein leakage (FL) test method OECD TG 460, revised in 2017 ESAC statement (2009); JaCVAM statement (2013)
Short time exposure (STE) in vitro test method OECD TG 491, revised in 2020 ICCVAM report (2013); JaCVAM statement (2016); KoCVAM guideline (2017)
Vitrigel-eye irritancy test (EIT) method OECD TG 494, revised in 2021
In vitro macromolecular test method OECD TG 496, published in 2019
Bovine corneal opacity and permeability (BCOP) test method OECD TG 437, revised in 2020 ICCVAM report (2006); ESAC statement (2007); JaCVAM statements (2009 and 2014); KoCVAM guideline (2011)
Isolated chicken eye (ICE) test method OECD TG 438, revised in 2018 ICCVAM report (2006); ESAC statement (2007); JaCVAM statement (2009)
Cytosensor microphysiometer (CM) assay  – ESAC statement (2009); ICCVAM report (2010)

Toxicity Endpoints: Skin Corrosion and Irritation

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Integrated approach on testing and assessment (IATA) for skin corrosion and irritation OECD GD 203, published in 2014
Chemical toxicity assessment strategy for skin corrosion and irritation European Chemicals Agency guidance Chapter R.7a., R.7.2 (2017)
In vitro membrane barrier test Corrositex for skin corrosion OECD TG 435, revised in 2015 ICCVAM report (1999); ESAC statement (2000); JaCVAM statement (2017)
In vitro skin corrosion: Reconstructed human epidermis (RhE) test (EpiSkin™, EpiDerm™, SkinEthic™, epiCS®, LabCyte EPI-MODEL24 SCT) OECD TG 431, revised in 2019 ICCVAM report (2002); ESAC statement (1998, 2000, 2006 2009); JaCVAM statement (2017)
In vitro skin corrosion: RhE test (Vitrolife-Skin™) JaCVAM statement (2008)
In vitro skin irritation: Reconstructed human epidermis (RhE) test (EpiSkin™, EpiDerm™, SkinEthic™, LabCyte EPI-MODEL24 SIT, Skin+®, epiCS®, KeraSkin) OECD TG 439, revised in 2021 ESAC statement (2007, 2008); JaCVAM statement (2010, 2013); KoCVAM guideline (2014, 2017)

Toxicity Endpoint: Skin Sensitization

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Adverse Outcome pathway (AOP) for skin sensitisation OECD GD 168 (Part 1, Part 2), published in 2012
Guideline on defined approaches on skin sensitisation OECD Guideline 497, published in 2021
Guidance on reporting of defined approaches and individual information sources to be used within
integrated approaches to testing and assessment (IATA)
OECD GD 256 (Annex I, Annex II), published in 2016
Use of alternative approaches for skin sensitisation as a replacement for animal testing US Environmental Protection Agency policy (2018)
Chemical toxicity assessment strategy European Chemicals Agency guidance Chapter R.7a., R.7.3.4–R.7.3.7 (2017)
OECD QSAR Toolbox (Implementing AOP workflow for skin sensitization, Example for predicting skin sensitisation of a mixture, Example of how to predict the skin sensitisation potential of a
chemical by read-across based on an analogue approach
)
OECD training manual, released in 2017
In chemico assays addressing the adverse outcome pathway (AOP) key event on covalent binding to proteins (direct peptide reactivity assay, DPRA; kinetic DPRA; amino acid derivative reactivity assay, ADRA) OECD TG 442C, revised in 2021 EURL ECVAM recommendation (2013); JaCVAM statement (2015); KoCVAM guideline (2016)
ARE-Nrf2 luciferase test method (KeratinoSens™, LuSens) OECD TG 442D, revised in 2018 EURL ECVAM recommendation (2014); JaCVAM statement (2015); KoCVAM guideline (2017)
In vitro assays addressing the adverse outcome pathway (AOP) key event on activation of dendritic cells (human cell line activation test, h-CLAT; IL-8 Luc assay; U937 skin sensitization test, U-SENS™) OECD TG 442E, revised in 2018 EURL ECVAM recommendation (2015); JaCVAM statement (2017); KoCVAM guideline (2017)

Toxicity Endpoint: Phototoxicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
3T3 neutral red uptake (NRU) phototoxicity test OECD TG 432, revised in 2019 ESAC statement (1997); ICH S10; KoCVAM guideline (2007)
Reactive oxygen species (ROS) assay OECD TG 495, published in 2019 JaCVAM statement (2016); ICH S10
Reconstructed human epidermis phototoxicity test method OECD TG 498, published in 2021 ICH S10

Toxicity Endpoint: Skin Absorption / Penetration

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
In vitro diffusion method OECD TG 428, published in 2004 JaCVAM statement (2014); KoCVAM guideline (2009)

Toxicity Endpoint: Acute systemic toxicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Guidance for waiving tests for pesticide formulations Canada Pest Management Regulatory Agency guidance (2013); US Environmental Protection Agency guidance for acute dermal toxicity tests (2016)
Strategy to replace, reduce, and refine the use of animals in the assessment of acute mammalian systemic toxicity EURL ECVAM guidance (2014)
Collaborative Acute Toxicity Modeling Suite (CATMoS) NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) (2019)
3T3 neutral red uptake (NRU) cytotoxicity test to identify substances not requiring classification EURL ECVAM recommendation (2013)

Toxicity Endpoint: Genotoxicity / Mutagenicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
OECD QSAR Toolbox: Example for predicting Ames mutagenicity using read-across OECD training manual, released in 2017
In vitro micronucleus test OECD TG 487, revised in 2016 ESAC statement (2006); ICH S2(R1)
Bacterial reverse mutation test OECD TG 471, revised in 2020 ICH S2(R1)
In vitro mammalian chromosome aberration test OECD TG 473, revised in 2016 ICH S2(R1)
In vitro mammalian cell gene mutation test OECD TG 476, revised in 2016
In vitro mammalian cell gene mutation tests using the thymidine kinase gene OECD TG 490, revised in 2016 ICH S2(R1)

Toxicity Endpoint: Carcinogenicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
In vitro cell transformation assays (CTA) OECD GD 214, published in 2015; OECD GD 231, published in 2016 EURL ECVAM recommendations (2012 and 2013)

Toxicity Endpoint: Pyrogenicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
In vitro monocyte activation tests (MAT) ICCVAM report (2008); ESAC statement (2006); European Pharmacopoeia general chapter 2.6.30; US Food and Drug Administration guidance (2012)

Toxicity Endpoint: Hematotoxicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
CFU-GM assay ESAC statement (2006)

Toxicity Endpoint: Reproductive toxicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Embryonic stem cell test ESAC statement (2001)
Micromass embryotoxicity assay (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.) ESAC statement (2001)
Whole rat embryotoxicity assay (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.) ESAC statement (2001)

Toxicity Endpoint: Endocrine disruptor screening

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Stably transfected transactivation in vitro assays to detect oestrogen receptor agonists and antagonists OECD TG 455, revised in 2021 JaCVAM statement (2016)
H295R steroidogenesis assay OECD TG 456, published in 2011
Stably transfected human androgen receptor transcriptional activation assay OECD TG 458, revised in 2020
Human recombinant oestrogen receptor (hrER) in vitro assays to detect chemicals with ER binding affinity OECD TG 493, published in 2015
Xenopus eleutheroembryonic thyroid assay (XETA) (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.) OECD TG 248, published in 2019

Toxicity Endpoint: Aquatic toxicity

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
OECD QSAR Toolbox: Example for predicting acute aquatic toxicity to fish of mixture with known components OECD training manual, released in 2017
EnviroTox database to calculate threshold values Health and Environmental Sciences Institute (2018)
Freshwater alga and cyanobacteria growth inhibition test OECD TG 201, published in 2011
Daphnia sp. acute immobilisation test OECD TG 202, published in 2004
Fish cell line acute toxicity – the RTgill-W1 cell line assay OECD TG 249, published in 2021 ISO 21115 standard (2019)
In vitro intrinsic clearance test using cryopreserved rainbow trout hepatocytes (Note: Animal primary cells are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test.) OECD GD 280, published in 2018
In vitro intrinsic clearance test using rainbow trout liver S9 sub-cellular fraction (Note: Animal primary cells are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test.) OECD TG 319B, published in 2018; OECD GD 280, published in 2018
Fish embryo acute toxicity test (FET) (Note: Animal embryos are used, therefore this test should be used only if replacing a regulatory requirement for a live-animal test using later life stages.) OECD TG 236, published in 2013 EURL ECVAM recommendation (2014)

Toxicity Endpoint: All endpoints

Test Methods and Approaches OECD Guidelines and Guidance Other Authority Recommendations
Guidance on considerations for waiving or bridging of mammalian acute toxicity tests OECD GD 237, published in 2016
Guidance for waiving or bridging of acute toxicity tests for pesticides Health Canada guidance (2013)
Guidance on the reporting of defined approaches to be used within integrated approaches to testing and assessment OECD GD 255, published in 2016
Guidance on the validation of (Q)SAR models OECD GD 69, published in 2007
OECD QSAR Toolbox: Guidance documents and training materials OECD, revised in 2018
QSAR Model Database Maintained by the European Commission Joint Research Centre
Various modelling programs For example, programs from Lhasa Limited, Leadscope, ScitoVation, and Simulations Plus
Guidance on the grouping of chemicals OECD GD 194, published in 2014  –
Read-across assessment framework European Chemicals Agency guidance (2017)
Guidance on good in vitro method practices OECD GD 286, published in 2018  –
Guidance for describing non-guideline in vitro test methods OECD GD 211, published in 2014  –
Classification of mixtures based on the toxicity of ingredients United Nations “Globally Harmonized System of Classification and Labelling of Chemicals” guidance (2015); US Environmental Protection Agency pilot program

Toxicity Endpoint: Biologics testing

Replacement Method or Strategy Regulatory acceptance
In vitro leptospirosis vaccine potency assay USDA Supplemental Assay Methods (SAM) 624, 625, 626 and 627
In vitro erysipelas vaccine potency assay USDA SAM 612 and 613
In vitro clostridial vaccine potency assay USDA draft SAM 220
In vitro tetanus toxoid potency assay USDA SAM 217
In vitro recombinant antibody production methods See www.thepsci.eu/antibodies/
Veterinary target animal batch safety test (TABST) Can be waived following demonstration of compliance; USDA CVB memorandum 800.116
Revocation of general safety tests (GST)/abnormal toxicity tests (ATT) FDA amended biologics regulations to revoke GST (2015); all European Pharmacopoeia monographs revised to revoke the ATT (2017)

For all endpoints, in vitro methods developed in-house should always be used.

Researchers should make every effort to use available non-animal methods. If these methods are not accepted by regulatory agencies, information on reduction and refinement methods can be found here:

Additional methods for reducing and replacing animal use are available in the following databases:

 Detailed information on the guidance documents and test methods described in this document can be found at the following sites: