Endpoint | Method Name | Test Type1 | Endorsement of Scientific Validity: Links to ESAC Statements; EURL ECVAM Recommendations; ECVAM Protocols; ICCVAM Evaluations; ICCVAM Protocols; JaCVAM Evaluations; JaCVAM Statements | Regulatory Acceptance | ||
Lead Authority | Subsequent Endorsement(s) | International (Links to OECD: OECD TGs; Draft TGs; GDs; Draft GDs) | National/ Regional2 | |||
Acute mammalian toxicity (oral) | Acute oral toxicity | In vivo | ESAC (2000) | OECD TG 401; Deleted in 2002 | ||
Acute toxic class method | In vivo | ESAC (2007) | OECD TG 423 (2001) | |||
Fixed dose procedure | In vivo | ESAC (2007) | OECD TG 420 (2001) | |||
Up-and-down procedure | In vivo | ICCVAM (2001) | ESAC (2007) | OECD TG 425 (2006) | EPA OPPTS 870.1100 (2002) | |
Normal human keratinocyte neutral red uptake (NHK NRU) assay | In vitro3 | ICCVAM (2008) | JaCVAM (2011) | OECD GD 129 (2010) | ||
Balb/c 3T3 neutral red uptake assay | In vitro3 | ICCVAM (2008) | JaCVAM (2011) | OECD GD 129 (2010) | ||
3T3 NRU Assay supporting identification of substances not requiring classification for acute oral toxicity | In vitro | EURL ECVAM (2013) | ||||
Waiving or bridging of mammalian acute toxicity tests30 | Draft OECD GD (2015) | |||||
Acute mammalian toxicity (hematotoxicity) | Colony Forming Unit-Granulocyte/Macrophage Assay for acute neutropenia in humans | In vitro | ESAC (2006) | |||
Acute mammalian toxicity (inhalation) | Acute toxic class method | In vivo | OECD TG 436 (2009) OECD GD No. 153 (2011) | |||
Fixed concentration procedure | In vivo | Draft OECD TG 433 (2015) | ||||
Acute toxicity testing of pesticides | Guidance for waiving or bridging of mammalian acute toxicity tests for pesticides (acute oral, dermal, inhalation; primary eye and dermal; dermal sensitization) | In vivo | EPA OPP (2012) | |||
Biologics & Vaccines | ELISA for swine erysipelas vaccines batch potency testing | In vitro | ESAC (2002) | ICCVAM agency: USDA (2009) | European Pharmacopeia, Monograph 064 (2004) | |
ELISA for human tetanus vaccines batch potency testing | In vitro | ESAC (2000) | European Pharmacopeia (2003) FDA, 21 CFR 610.10 (accepted on case-by-case basis) | |||
Toxin binding inhibition test for human tetanus vaccines batch potency testing | In vitro | ESAC (2000) | European Pharmacopeia (2003) FDA, 21 CFR 610.10 (accepted on case-by-case basis) | |||
Deletion of target-animal batch safety test (TABST) for batch safety testing of veterinary vaccines after consistency in 10 consecutive batches | NA | ESAC (2002) | VICH GL50 (2013) | European Pharmacopeia (2012) | ||
Vaccines for Veterinary Use (0062): around 80 vaccine-specific monographs with VICH Guidelines; around 40 inactivated vaccine-specific monographs | VICH 41; VICH 44 | European Pharmacopeia (2012) European Pharmacopoeia, 2015 (effective January 1, 2017) | ||||
ELISA for in vitro batch potency testing of Leptospira veterinary vaccines | In vitro | ICCVAM agency: USDA (2008) | USDA Supplemental Assay Methods 624, 625, 626, and 627 | |||
Use of humane endpoints in animal testing of veterinary biologics, including rabies vaccines | In vivo | ICCVAM agency: USDA (2004; 2012) | OECD GD No. 19 | USDA: 9 CFR Part 113, Notice No. 04-09 (2004); 9 CFR 113, CVB Notice No. 12-12 | ||
Veterinary vaccine potency assays: exemptions from Standard Requirements tests; master reference qualification and requalification | NA | ICCVAM agency: USDA | USDA: 9 CFR 113.409(c), Memorandum 800.114 (2012) | |||
Cell-based assay for stability and potency of botulinum neurotoxin type A products15a | In vitro | ICCVAM agency: FDA ICCVAM Workshop (2006) | US FDA: Allergan, Inc., method accepted (2011) | |||
Cell-based assay for stability and potency of botulinum neurotoxin type A products15b | In vitro | US FDA (2015) German Federal Institute for Drugs and Medical Devices and all other EU drug regulatory authorities (2015) | ||||
Alternative test procedure for tuberculin, PPD Bovis, intradermic | In vivo | ICCVAM agency: USDA | USDA: 9 CFR 113.409(c), Memorandum 800.114 (2012) | |||
Carcinogenicity | Cell Transformation Assays (CTAs) in: Syrian Hamster Embryo cells (SHE) performed at pH 6.7 and at pH 7.04 | In vitro | EURL ECVAM (2012) | OECD Detailed Review Paper No. 31 OECD GD No. 214 (2015) | ||
Bhas 42 Cell Transformation Assay (Bhas 42 CTA)4 | In vitro | EURL ECVAM (2013) | OECD Validation study report no. 208 (2014) OECD GD No. 231 (2016) | |||
Chronic toxicity | Ending 1-year dog studies of pesticides | In vivo | ESAC (2006) | Revised US EPA Pesticide Data Requirements | ||
Dermal absorption/penetration | In vitro skin absorption methods | In vitro | OECD Expert Group (2002) | JaCVAM (2013) | OECD TG 428 (2004) OECD GD 28 (2004) OECD Guidance Notes No. 156 | |
Ecotoxicity | Acute aquatic toxicity: Upper threshold concentration step-down approach | In vivo | ESAC (2006) | OECD GD 126 (2010) | EU Biocides Regulation and REACH Guidance | |
Acute avian toxicity (oral): Sequential testing procedure to minimize numbers of birds used | In vivo | OECD TG 223 (2010; updated 2016) | ||||
Zebrafish embryo acute toxicity test (ZFET)18 | In vivo | EURL ECVAM (2014) | OECD TG 236 (2013) | |||
Honey bee larval toxicity test, repeated dose29 | In vivo | Draft OECD GD (2015) | ||||
Endocrine active substances | OECD Detailed Review Paper No. 97 (2008) | |||||
Androgen receptor binding assay (rat prostate cytosol) | Ex vivo | OPPTS TG 890.1150 (EPA, 2009) | ||||
Aromatase inhibition assay (human recombinant) | In vitro | OPPTS TG 890.1200 (EPA, 2009) | ||||
Performance-based Test Guideline for stably transfected human receptor-alpha transcriptional activation assays for detection of estrogenic agonist-activity of chemicals | In vitro | OECD/US EPA | OECD TG 455 (2012; updated 2015; updated 2016)5 | OPPTS TG 890.1300 | ||
Estrogen Receptor binding assay Rat Uterine Cytosol (ER-RUC) | Ex vivo | OPPTS TG 890.1250 (EPA, 2009) | ||||
H295R steroidogenesis assay5 | In vitro | OECD/US EPA | OECD TG 456 (2011) | OPPTS TG 890.1550 (EPA, 2009) | ||
US EPA Endocrine Disruptor Screening Program | In vitro / In vivo
In vitro | US EPA Tier 1 Screening Battery (2009)5 US EPA Toxcast™ ER Model for Bioactivity (2015) | ||||
BG1Luc ER TA test method for estrogen agonists and antagonists6 | In vitro | ICCVAM (2012) | JaCVAM (2013) | OECD TG 457 (2012) Draft Performance Standards for TG 457 (2012) | ||
Performance-based Test Guideline for human recombinant estrogen receptor (hrER) in vitro assays to detect chemicals with ER binding affinity | OECD TG 493 (2015)21 | |||||
Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for detection of androgenic agonist and antagonist activity of chemicals | OECD TG 458 (2016) | |||||
Eye corrosion/Serious eye damage | Bovine Corneal Opacity Permeability (BCOP) test34 | Ex vivo | ICCVAM (2007) | ESAC (2007) JaCVAM (2009) | OECD TG 437 (2009; updated 2013) | REACH regulation |
Isolated Chicken Eye (ICE) test | Ex vivo | ICCVAM (2007) | ESAC (2007) JaCVAM (2009) | OECD TG 438 (2009; updated 2013; draft revised 2015)10 | REACH regulation | |
Cytosensor Microphysiometer | In vitro | ESAC (2009)7 | ICCVAM (2010)8 | Draft OECD TG (2012) | ||
Fluorescein Leakage | In vitro | ESAC (2009)9 | ICCVAM
JaCVAM (2012) | OECD TG 460 (2012) OECD Summary Document No. 180 | REACH regulation | |
Hen’s Egg Test-ChorioAllantoic Membrane (HET-CAM) | In vitro | EU Competent Authorities for Dangerous Substances Directive | ||||
Isolated rabbit eye test | Ex vivo | EU Competent Authorities for Dangerous Substances Directive | ||||
Short Time Exposure (STE) in vitro test method for identifying chemicals inducing serious eye damage | In vitro | JaCVAM | ICCVAM (2013) | OECD TG 491 (2015) | REACH regulation | |
Routine use of topical anesthetics, systemic analgesics, and humane endpoints | In vivo | ICCVAM (2010) | JaCVAM (2014) | OECD TG 405 (updated 2012) OECD GD No. 19 | ||
Sequential testing strategy for eye irritation and corrosion | In vitro/Ex vivo/In vivo | ICCVAM16 | JaCVAM (2014) | OECD TG 405 (updated 2012) | ||
Alternate testing framework for classification of eye irritation potential of EPA pesticide products28 | In vitro | EPA OPP (2015) | ||||
Eye irritation | Cytosensor Microphysiometer | In vitro | ESAC (2009)7 | ICCVAM (2010)8 | Draft OECD TG (2012) | |
Short Time Exposure (STE) in vitro test method for identifying chemicals not requiring classification for eye irritation or serious eye damage | In vitro | JaCVAM | ICCVAM (2013) | OECD TG 491 (2015) | REACH regulation | |
Reconstructed Human Cornea-like Epithelium (RhCE) test method for identifying chemicals not requiring classification and labelling for eye irritation or serious eye damage22 | In vitro | EURL ECVAM | Draft Revised OECD TG 492 (2016) Draft updated performance standards for validation of similar methods (2016) | REACH regulation | ||
Bovine Corneal Opacity Permeability (BCOP) Test | Ex vivo | ICCVAM (2007) | ESAC (2007) JaCVAM (2013) | OECD TG 437 (2009; updated 2013)10 OECD Summary Document 189 (2013) OECD GD 160 | REACH regulation | |
Isolated Chicken Eye (ICE) test | Ex vivo | ICCVAM (2007) | ESAC (2007) JaCVAM (2014) | OECD TG 438 (2009; updated 2013; draft revised 2015)10 Draft Position Paper on ICE for UN GHS No Category OECD Summary Document 188 Part 1 (2013); 188 Part 2 (2013) OECD GD 160 | REACH regulation | |
Routine use of topical anesthetics, systemic analgesics, and humane endpoints | In vivo | ICCVAM (2010) | JaCVAM (2014) | OECD TG 405 (updated 2012) OECD GD No. 19 | ||
Sequential testing strategy for eye irritation and corrosion | In vitro/Ex vivo/In vivo | ICCVAM16 | JaCVAM (2014) | Updated OECD TG 405 (2012) | ||
Alternate testing framework for classification of eye irritation potential of EPA pesticide products28 | In vitro | EPA OPP (2015) | ||||
Genotoxicity25 | Bacterial reverse mutation (Ames) test | In vitro | OECD TG 471 (1997) | |||
In vitro mammalian cell gene mutation tests using the Hprt and xprt genes | In vitro | OECD TG 476 (1997; updated 2015; updated 2016) | ||||
In vitro mammalian cell gene mutation test using the thymidine kinase gene | In vitro | OECD TG 490 (2015; updated 201619 | ||||
In vitro mammalian chromosomal aberration test | In vitro | OECD TG 473 (1983; updated 2014; updated 2016) OECD Report on Statistical Issues…(No. 198) | ||||
In vitro mammalian cell micronucleus test | In vitro | ESAC (2006) | ICCVAM16 | OECD TG 487 (2010; updated 2014; updated 2016) ICH (2011) OECD Report on Statistical Issues…(No. 198) | ||
In vitro sister chromatid exchange test | In vitro | OECD TG 479 (1986; Deleted in 2014) | ||||
In vitro unscheduled DNA synthesis test | In vitro | OECD TG 482 (1986; Deleted in 2014) | ||||
Saccharomyces cerevisiae gene mutation assay | In vitro | OECD TG 480 (1986; Deleted in 2014) | ||||
Saacharomyces cerevisiae mitotic recombination assay | In vitro | OECD TG 481 (1986; Deleted in 2014) | ||||
In vivo mammalian alkaline comet assay | In vitro | JaCVAM | OECD TG 489 (2016) | |||
Metabolism | Human cytochrome P450 (CYP) n-fold induction in vitro test method | In vitro | Draft OECD TG xxx (2014) Draft Performance Standards (2014) | |||
Human drug-drug interactions: pathway identification; enzyme inhibition; enzyme induction; transporter substrate identification; transporter inhibition | In vitro | US FDA (2012) EMA (2013) | ||||
The use of metabolizing systems for in vitro testing of endocrine disruptors | OECD Detailed Review Paper No. 97 (2008) | |||||
Phototoxicity | 3T3 Neutral Red Uptake Phototoxicity Test33 | In vitro | ESAC (1997) | OECD TG 432 (2004) | ||
3T3 NRU Phototoxicity Test: Application to UV filter chemicals | In vitro | ESAC (1998) | OECD TG 432 (2004) | |||
(ROS) Reactive Oxygen Species assay photosafety (non-biological method)23 | In vitro | JaCVAM26 (2013) | ICH S10 (2013) Draft OECD TG (2016) | |||
Preclinical and nonclinical safety studies for drug development (harmonized guidance can reduce use of animals) | Guidance on nonclinical safety studies for the conduct of human clinical trials and marketing authorization for pharmaceuticals M3(R2) | In vivo | ICH (2010) | |||
Preclinical safety evaluation of biotechnology-derived pharmaceuticals, ICH S6(R1) | In vivo | ICH (1997; updated 2011) | ||||
Pyrogenicity | Five human cell assays for detecting pyrogenicity produced by Gram-negative endotoxins27: Human whole blood IL-1; Human whole blood IL-6; Human cryopreserved whole blood IL-1; PBMC IL-6; PBMC IL-6 | In vitro | ESAC (2006) | ICCVAM (2008)12 | European Pharmacopeia, Monocyte activation test (2.6.30) (2010); revised (2.6.30) (July 2017) FDA Guidance: Pyrogen and Endotoxins Testing (2012) | |
Limulus amebocyte lysate (LAL) test | In vivo/In vitro | EU, US, and Japanese Pharmacopeia (2005) ICH harmonized text (2010) | ||||
Reproductive & developmental toxicity | Embryonic stem cell test for embryotoxicity17 | In vitro | ESAC (2002) | |||
Micromass embryotoxicity assay | Ex vivo | ESAC (2002) | ||||
Whole rat embryotoxicity assay | Ex vivo | ESAC (2002) | ||||
Extended one-generation reproductive toxicity study | In vivo | ICCVAM16 | OECD TG 443 (updated 2012) | |||
Skin corrosion | In vitro skin corrosion: reconstructed human epidermis (RHE) test method: Episkin™ SCT, Epiderm™ SCT, SkinEthic™ RHE SCT, epiCS® SCT (formerly EST-1000); Vitrolife-Skin™ | In vitro | ESAC (1998; 2000; 2006; 2009) JaCVAM (2008) | ICCVAM 2002 | OECD TG 431 (2004; updated 2015; updated 2016)13a OECD Summary Document 190 (2013) | REACH regulation |
In vitro Membrane Barrier test method for skin corrosion: CORROSITEX™ | In vitro | ICCVAM (1999) | ESAC (2000) | OECD TG 435 (2006; updated 2015) | REACH regulation | |
Rat skin transcutaneous electrical resistance (TER) test method | Ex vivo | ESAC (1998) | ICCVAM (2002) | OECD TG 430 (2004; updated 2015) | REACH regulation | |
Integrated approach on testing and assessment (IATA) for skin corrosion and irritation | OECD GD 203 (2014) | |||||
Skin irritation | Reconstructed human epidermis test method: Episkin™ SIT, Epiderm™ SIT, SkinEthic™ RHE SIT, LabCyte EPI-MODEL24 SIT | In vitro | ESAC (2007; 2008; 2009) JaCVAM (2013) | JaCVAM (2010; 2012) | OECD TG 439 (2010; updated 2015)13b | REACH regulation |
Integrated approach on testing and assessment (IATA) for skin corrosion and irritation | OECD GD 203 (2014) | |||||
Skin sensitization | Local lymph node assay (LLNA) | In vivo | ICCVAM (1999; 2009) | ESAC (2000) | OECD TG 429 (2002; updated 2010) | |
Reduced LLNA: rLLNA11 | In vivo | ESAC (2007; 2008) | ICCVAM (2009) JaCVAM (2012) | OECD TG 429 (updated 2010) | ||
Nonradiolabelled LLNA: DA | In vivo | ICCVAM (2010) | JaCVAM (2012) | OECD TG 442A (2010) | ||
Nonradiolabelled LLNA: BrdU-ELISA | In vivo | ICCVAM (2010) | JaCVAM (2012) | OECD TG 442B (2010) | ||
LLNA for Potency Categorization of Skin Sensitizers | In vivo | ICCVAM (2011) | UN GHS (2009) | Some US agencies (2012) | ||
ARE-Nrf2 luciferase test method20a: KeratinoSens™ | In vitro | EURL ECVAM (2014) | JaCVAM (2015) | OECD TG 442D (2015) | REACH regulation | |
Direct Peptide Reactivity Assay (DPRA)20b | In chemico | EURL ECVAM (2013) | JaCVAM (2015) | OECD TG 442C (2015) | REACH regulation | |
Human Cell Line Activation Test (h-CLAT)20c | In vitro | EURL ECVAM (2015) | OECD TG 442E (2016) | REACH regulation | ||
IL-8 Luc Assay32 | In vitro | JaCVAM | Draft OECD TG (2016) | |||
U937 Skin Sensitization Test (U-SENS™)31 | In vitro | EURL ECVAM (2016) | Draft OECD TG (2016) | |||
P&G/NICEATM Open-source Integrated Testing Strategy to identify potential skin sensitizers without conducting animal tests | In vitro | NICEATM (2015) | ||||
Reporting of Defined Approaches and Individual Information Sources to be used within Integrated Approaches to Testing and Assessment (IATA) for Skin Sensitisation- ENV/JM/HA(2016)11 | OECD GD 256 (2016) Annex 1: Case Studies… (2016) Annex II: Information used within the case studies…(2016) |
1 Table content is limited to in vitro, ex vivo, in chemico and/or in silico test methods, and in vivo methods or test schemes proposed to reduce or refine animal use.
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 In vitro cell transformation assays (CTAs) can detect both genotoxic and non-genotoxic carcinogens, which can be distinguished by parallel use of genotoxicity test(s). CTA results are recommended for use as part of a testing strategy and/or weight-of-evidence approach for the assessment of carcinogenic potential, and may reduce the use of in vivo testing.
5 The two reference test methods that provide the basis for this guideline are: the Stably Transfected TA (STTA) assay using the (h) ERα-HeLa-9903 cell line, and the BG1Luc ER TA assay. These methods are proposed for screening and prioritization purposes, but also provide mechanistic information that can be used in a weight of evidence approach.
6 BG1Luc ER TA test method has been adapted to a high throughput screening (HTS) format and 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 BCOP and ICE methods can be used to identify chemicals as either 1) causing “serious eye damage” (category 1 of GHS, or 2) not requiring classification for eye irritation or serious eye damage according to GHS (new to the 2013 TG updates).
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)
13a Four validated test methods using commercially available RhE models are included in TG 431: EpiSkin™ Standard Model (SM), EpiDerm™ Skin Corrosivity Test (SCT) (EPI-200), SkinEthic™ RHE, and epiCS® (previously named EST-1000).
13b Four validated test methods adhere to TG 439: EpiSkin™, EpiDerm™ SIT (EPI-200), SkinEthic™ RHE, and LabCyte EPI-MODEL24 SIT. “Depending on the regulatory framework and the classification system in use, this procedure may be used to determine the skin irritancy of test substances as a stand-alone replacement test for in vivo skin irritation testing, or as a partial replacement test, within a tiered testing strategy.”
14 Recommended as screening test or as part of sequential testing strategy; only positive test results accepted in the 2007 endorsement
15a Cell-based assay for botulinum toxin potency testing is limited to products produced using the Allergan method
15b Cell-based assay for botulinum toxin type A testing is limited to products produced by Merz
16 Contributed to OECD TG development and/or review
17 Draft OECD GD 43, see page 18
18 EURL ECVAM reports that: “TG236 does not indicate whether the fish embryo acute toxicity test can be used as an alternative to the OECD TG203; however, several recently published papers demonstrate that the LC50 values produced with the fish embryo acute toxicity test correlate well with those observed in juvenile or adult fish (Lammer et al, 2009; Knoebel et al 2012; Belanger et al (2013).”
19 The assays using the thymidine kinase (TK) locus were originally contained in TG 476.
20 In vitro and in chemico methods endorsed as valid for supporting the discrimination between skin sensitizers and non-sensitizers in accordance with the UN GHS. The guidelines explain it is likely that combinations of non-animal methods using integrated testing approaches will be needed to substitute for the animal tests.
20a In vitro method endorsed as valid for supporting the discrimination between skin sensitizers and non-sensitizers in accordance with the UN GHS. The guidelines explain it is likely that combinations of non-animal methods using integrated testing approaches will be needed to substitute for the animal tests. The only ARE-Nrf2 luciferase test method covered currently by this TG, is the KeratinoSens™ test method.
20b In chemico method endorsed as valid for supporting the discrimination between skin sensitizers and non-sensitizers in accordance with the UN GHS. The guidelines explain it is likely that combinations of non-animal methods using integrated testing approaches will be needed to substitute for the animal tests.
20c In vitro method endorsed as valid for supporting the discrimination between skin sensitizers and non-sensitizers in accordance with the UN GHS. The guidelines explain it is likely that combinations of non-animal methods using integrated testing approaches will be needed to substitute for the animal tests.
21 This guideline describes assays providing the methodology for human recombinant in vitro assays to detect substances with estrogen receptor binding affinity, and comprises two similar reference test methods: Freyberger-Wilson (FW) in vitro estrogen receptor binding assay…, and Chemical Evaluation and Research Institute (CERI) in vitro estrogen receptor binding assay…; proposed for screening and prioritization purposes, and provide mechanistic information that can be used in a weight of evidence approach.
22 Methods currently covered by this TG are the EpiOcular™ Eye Irritation Test (EIT) and the SkinEthic™ Human Corneal Epithelium (HCE) Eye Irritation Test (EIT).
23 ROS assay can be used in an integrated photosafety testing and decision strategy for drug research and development where negative results would not require further testing, while positive and inconclusive results would proceed to the next level of testing in an in vitro test system such as the 3T3 Phototoxicity Assay.
24 OECD Series on Testing and Assessment contains additional useful guidance and performance standards. For example, Guidance Document No. 211: Guidance for describing non-guideline in vitro test methods to facilitate their consideration in regulatory applications, and the draft Guidance Document on Revisions to OECD Genetic Toxicology Test Guidelines.
25 Also see: Draft Guidance Document on Revisions to OECD Genetic Toxicology Test Guidelines (2015) and Report on Statistical Issues Related to OECD Test Guidelines (TGs) on Genotoxicity (Series on Testing and Assessment, No. 198, 2014)
26 “The JaCVAM peer review panel concluded that the reproducibility and predictivity of the ROS assay is sufficient to support its use in an integrated photosafety testing and decision strategy for drug research and development…[where] negative results in the ROS assay would not require further testing in animals or other tests, while positive, weakly positive, and inconclusive results would proceed to the next level of testing in an in vitro test system such as the 3T3 Phototoxicity Assay (OECD Test Guideline 432).”
27 The in vitro pyrogenicity methods are not considered valid for use in Japan at this time.
28 Includes explanation of expanded use of BCOP for antimicrobial testing, and decision tree approach using 3 assays (see IIVS Decision Tree Approach):
29 Honey bee larval toxicity tests complement OECD TGs 213 and 214 on young adult honey bees and should be seen as a lower tier screening test.
30 “Generally, waivers are considered when there is little or no significant human exposure by a given route of exposure or when it is technically not possible to perform a study for a certain endpoint, such as not requiring an acute oral toxicity study when the test chemical exists as a vapour or gas. Waivers are also possible taking into account animal welfare considerations, such as when the test chemical is corrosive.”
31 U-SENS™ is proposed to address the third key event of the skin sensitization AOP; recommended by EURL ECVAM as part of an IATA to support discrimination between sensitizers and non-sensitizers for hazard classification and labelling.
32 “IL-8 Luc assay is proposed to address the third key event (dendritic cell activation) of the skin sensitisation AOP by quantifying changes in the expression of cytokine associated with the process of activation of DC (i.e. IL-8), in the human monocytic leukemia cell line THP-1-derived IL-8 reporter cell line, THP-G8, following exposure to sensitisers.”
33 EPAA-IIVS Video of 3T3 NRU Phototoxicity Assay.
34 EPAA-IIVS Video of BCOP Eye Irritation Assay.
A. EURL ECVAM: Test methods endorsed by EURL 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; 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. OECD Guidance Document for Describing Non-Guideline In Vitro Test Methods (No.211): intended to harmonize the way non-guideline in vitro test methods are described.
F. OECD Guidance Document on Developing and Assessing Adverse Outcome Pathways (No. 184) (2013): intended to provide guidance on the development of Adverse Outcome Pathways (AOPs).
G. OECD Guidance Document on the Reporting of Defined Approaches to be used within Integrated Approaches to Testing and Assessment (No. 255) (2016): intended “to provide a set of principles for reporting defined approaches to testing and assessment to facilitate their evaluation. Templates are also provided to enable a structured and harmonised approach to their documentation.”
H. ICH test guidelines: technical requirements for medicinal products containing new drugs accepted by the regulatory bodies of EU, Japan, and USA
I. VICH test guidelines: EU-Japan-USA programme aimed at harmonising technical requirements for veterinary product registration
J. 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)
K. USDA APHIS Supplemental Assay Methods (SAMs) (updated March 17, 2016)
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
IATA: Integrated Approach to Testing and Assessment, “Approach based on multiple information sources that integrates and weights all relevant existing evidence and guides the targeted generation of new data, where required, to inform regulatory decision-making regarding potential hazard and/or risk.”
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
Author(s)/Contributor(s):
Sherry L. Ward, PhD, MBA
AltTox Contributing Editor
AltTox Editorial Board reviewer(s):
Ian Kimber, PhD
Professor, University of Manchester
João Barroso, PhD
Systems Toxicology Unit, EU Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM)
Albert P. Li, PhD
President & CEO, In Vitro ADMET Laboratories LLC
Disclaimer
The information provided here is intended only as an overview, and is neither guidance or a comprehensive review of the laws, regulations, or toxicity tests that may be accepted by an individual agency. Countries/regions and their regulatory authorities usually provide specific guidance on hazard/toxicity testing requirements.
The definitive resource for non-animal chemical assessment methods.
© 2019 The Humane Society of the United States and the Procter & Gamble CompanyWeb Design by Zen Den