ICCVAM-Recommended Test Method Protocols

This page contains links to current ICCVAM-recommended protocols for specific test methods. It also includes links to the ICCVAM Test Method Evaluation Reports where the protocols were originally published and to pages on this website that contain additional information about the evaluation of the methods.

Acute Systemic Toxicity

Revised Up-and-Down Procedure for Acute Oral Toxicity Testing

In Vitro Cytotoxicity Test Methods for Estimating Starting Doses for Acute Oral Systemic Toxicity Tests

Endocrine Disruptors

Protocols for the BG1Luc Estrogen Receptor Transactivation Assays

Note: Beginning in 2016, the BG1Luc ER TA assay will be referred to as the VM7Luc ER TA assay. This reflects new information regarding the identity of the cell line used in the assay. Refer to the webpage summarizing the assay validation for more details.

Eye Corrosion and Irritation

In Vitro Test Methods for Assessing Potential to Cause Chemically Induced Eye Injuries

The most recent ICCVAM evaluation of these test methods was published in the ICCVAM Test Method Evaluation Report, NIH Document No. 10-7553, 2010.

Note: Some of the protocols listed are recommended only for non-regulatory, validation, or optimization studies. Such limitations are noted on the title pages of the relevant protocols.

Pyrogenicity

In Vitro Test Methods Proposed for Assessing Pyrogenicity of Pharmaceuticals and Other Products

Protocols were published as Appendix C to ICCVAM Test Method Evaluation Report, NIH Document No. 08-6392.

Note: Availability of the Mono Mac 6 cell line for commercial testing purposes may be limited. Go to the in vitro pyrogen test method evaluation page for details.

Skin Sensitization

In Vitro Test Methods for Identifying Potential Skin Sensitizers

These two assays were evaluated by the European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM). In a 2014 statement, ICCVAM concurred with EURL ECVAM that the methods can be used as part of an integrated testing strategy to identify potential skin sensitizers, but emphasized that neither method should be used as a standalone method for the identification of potential skin sensitizers or to determine skin sensitization potency categories. Test guidelines describing the methods were accepted in 2015 by the Organisation for Economic Co-operation and Development (OECD).

Updated Protocol for the Murine Local Lymph Node Assay (LLNA) for Assessing Allergic Contact Dermatitis Hazard Potential

This protocol is also recommended for use with the reduced LLNA. Learn more about the ICCVAM evaluation of the reduced LLNA.

Nonradiolabeled Murine Local Lymph Node Assay (LLNA) Methods for Assessing Allergic Contact Dermatitis Hazard Potential

Note: In guidance issued in February 2020, FDA stated that it no longer recommends that sponsors conduct the murine local lymph node assay to assess the sensitization potential of topical drug products due to the limitations of the assay. As an alternative to accepted guinea pig tests, FDA will consider a battery of in silico, in chemico, and in vitro studies that have been shown to adequately predict human skin sensitization with an accuracy similar to existing in vivo methods.