National Toxicology Program

This page has links to current ICCVAM-recommended protocols for the listed methods, the ICCVAM Test Method Evaluation Reports in which the protocols were originally published, and pages on the NICEATM-ICCVAM website on which additional information about the evaluation of the method can be found.

Please Note: Many of the complete ICCVAM Test Method Evaluation Reports are large documents and may take a few minutes to download.

Select links to jump to test method area of interest:

Acute Systemic Toxicity

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

• Protocol for the Revised Up-and-Down Procedure [PDF]
  Published as Appendix B to ICCVAM Test Method Evaluation Report, NIH Document No. 02-4501, 2001
  Volume 1 of NIH 02-4501 [PDF]
  Volume 2 of NIH 02-4501 [PDF]
  More information about the ICCVAM evaluation of the up-and-down procedure

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

• Protocol for the BALB/c 3T3 NRU Cytotoxicity Test Method [PDF]
  Protocol for the NHK NRU Cytotoxicity Test Method [PDF]
  Published as Appendix C to ICCVAM Test Method Evaluation Report, NIH Document No. 07-4519, 2006 - View report
  More information about the ICCVAM evaluation of in vitro cytotoxicity test methods

Immunotoxicity: Allergic Contact Dermatitis

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

• Updated Protocol for the Murine Local Lymph Node Assay (LLNA) [PDF]
  Published as Appendix A to ICCVAM-recommended Performance Standards for the LLNA, NIH Document No. 09-7357, 2009 [PDF]
  More information about development of performance standards for the LLNA
• This protocol is also recommended for use with the reduced LLNA
  More information about the ICCVAM evaluation of the reduced LLNA

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

• Protocol for the LLNA: DA [PDF]
  Published as Appendix B to ICCVAM Test Method Evaluation Report, NIH Document No. 10-7551, 2010 [PDF]


• Protocol for the LLNA: BrdU-ELISA [PDF]
  Published as Appendix B to ICCVAM Test Method Evaluation Report, NIH Document No. 10-7552, 2010 [PDF]


• More information about the ICCVAM evaluation of nonradiolabeled LLNA methods

Ocular Toxicity

In Vitro Test Methods for Assessing Potential to Cause Chemically Induced Eye Injuries
Please note: Some of the protocols listed below are recommended only for nonregulatory, validation, or optimization studies. Such limitations are noted on the title pages of the relevant protocols.

• Protocol for the Bovine Corneal Opacity and Permeability Test Method Revised March 2011 [PDF]
  Please Note: In the revised protocol, the examples of positive control substances have been updated and clarified and the intended purpose of the positive control in the assay has been clarified (see Section 5.3 [Positive Control]).
  Protocol for the Cytosensor Microphysiometer Test Method [PDF]
  Protocol for the Hen's Egg Test—Chorioallantoic Membrane Test Method [PDF]
  Protocol for the Isolated Chicken Eye Test Method Revised December 2010 [PDF]
  Protocol for the Isolated Rabbit Eye Test Method [PDF]
  Protocols originally published as Appendix B to ICCVAM Test Method Evaluation Report, NIH Document No. 10-7553, 2010
  Volume 1 of NIH 10-7553 [PDF]
  Volume 2 of NIH 10-7553 [PDF]
  More information about the ICCVAM evaluation of in vitro ocular testing methods


• Protocol for the EpiOcular Test Method [PDF]
  Published as Appendix B3 to ICCVAM Test Method Evaluation Report, NIH Document No. 10-7513, 2010 [PDF]
  More information about the ICCVAM evaluation of an in vitro testing strategy for ocular hazard classification and labeling of antimicrobial cleaning products

Routine Use of Topical Anesthetics, Systemic Analgesics, and Humane Endpoints to Avoid or Minimize Pain and Distress in Ocular Safety Testing

• Protocol: ICCVAM Recommendations for Revisions to OECD Test Guideline 405: Acute Eye Irritation/Corrosion [PDF]
  Published as Appendix B to ICCVAM Test Method Evaluation Report, NIH Document No. 10-7514, 2010 [PDF]
  More information about the ICCVAM assessment of the use of anesthetics, analgesics, and humane endpoints in ocular safety testing

Pyrogenicity

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

• Protocol for the Human Whole Blood/Interleukin-1β Test Method [PDF]
  Protocol for the Human Whole Blood/Interleukin-1β Test Method: Application of Cryopreserved Human Whole Blood [PDF]
  Protocol for the Human Whole Blood/Interleukin-6 Test Method [PDF]
  Protocol for the Human Peripheral Blood Mononuclear Cell/Interleukin-6 Test Method [PDF]
  Protocol for the Mono Mac 6/Interleukin-6 Test Method [PDF]
  Published as Appendix C to ICCVAM Test Method Evaluation Report, NIH Document No. 10-7553, 2010 [PDF]
  More information about the ICCVAM evaluation of in vitro pyrogen testing methods

Please note: Availability of the Mono Mac 6 cell line for commercial testing purposes may be limited. See the in vitro pyrogen test method evaluation page of the NICEATM-ICCVAM website for details.