Alternatives in Toxicology

U.S. and International Acceptance of Alternative Methods, 1998-2012

This table presents a chronological list of alternative test methods.


Guidance for Industry – Pyrogen and Endotoxins Testing: Questions and Answers (PDF | 174 KB)

DHHS. Food and Drug Administration.

June 2012- Includes guidance regarding the use of alternatives to the rabbit pyrogen test (RPT) and the bacterial endotoxins test (BET) for detecting pyrogens in pharmaceuticals and other products. Examples of alternative assays cited include the recombinant horseshoe crab Factor C asssay and monocyte activation type pyrogen tests (MATs).


Seurat-1: Towards the replacement of in vivo repeated dose systemic toxicity testing

The first step in a European research initiative to address the long term strategic target of "Safety Evaluation Ultimately Replacing Animal Testing (SEURAT)".


NTP Requests Information on Assays and Approaches for Neurotoxicity Screening (PDF | 197.3 KB)

Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM)

The National Toxicology Program (NTP) requests information on medium- or high-throughput technologies/assay systems, which allow for the batch screening of compounds (e.g., 25-50) in biochemical- or cell-based assays or alternative (non-rodent) animal models, that might be used to prioritize compounds for in vivo neurotoxicity testing.

The deadline for receipt of information is April 30. More information about the request for information was published April 1 in the Federal Register.


Leptospira workshop poster Proceedings: Workshop on Alternative Methods for Leptospira Vaccine Potency Testing

National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM).


ICCVAM Evaluation of the Use of the LLNA for Potency Categorization

Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM)

The murine local lymph node assay (LLNA) is a stand-alone test method for assessing the hazard potential of a test substance to induce allergic contact dermatitis (ACD). ICCVAM evaluated the use of the LLNA for potency categorization of chemicals causing ACD in humans. ICCVAM concluded that the LLNA can be used to categorize substances as strong sensitizers (Globally Harmonized System of Classification and Labelling of Chemicals Subcategory 1A), although it cannot be considered a stand-alone assay to categorize substances as other sensitizers (Globally Harmonized System of Classification and Labelling of Chemicals Subcategory 1B). Strong sensitizers are those substances considered to have a significant potential for causing hypersensitivity.


TSAR: Tracking System for Alternative test methods Review, Validation and Approval in the Context of EU Regulations on Chemicals

European Commission. Joint Research Centre. Institute for Health and Consumer Protection.

A tool to provide information on the status of alternative methods as they progress from scientific protocols to validation for being used in a regulatory context.

Access: free online


ICCVAM Recommendations to Provide for More Humane Eye Safety Testing (PDF | 149KB)

Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM)

Four test method evaluation reports are now available that provide recommendations for performing eye safety testing more humanely and for developing and using non-animal test methods and strategies.


Reduced Murine Local Lymph Node Assay

The National Toxicology Program - Interagency Center for the Evaluation of Alternative Toxicological Methods

The National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM), on behalf of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), announces recommendations for the reduced murine Local Lymph Node Assay (LLNA).


Dots part of the logo image for AltTox.org AltTox.org

Humane Society of the United States and Proctor & Gamble.

The goal of this web site is to advance non-animal methods of toxicity testing through online discussion and information exchange.


Final Recommendations on In Vitro Test Methods for Detecting Ocular Corrosives and Severe Irritants

The National Toxicology Program - Interagency Center for the Evaluation of Alternative Toxicological Methods

The National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM), on behalf of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), announces the availability of this report that describes and evaluates four ocular toxicity test methods evaluated by ICCVAM.


Guidance Document on Using In Vitro Data to Estimate In Vivo Starting Doses for Acute Toxicity

DHHS. NIH. National Institute of Environmental Health Sciences. National Institutes of Health.

Describes how to use in vitro cytotoxicity tests to estimate starting doses for acute oral lethality assays.


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

NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) and the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM).

The recommendations include protocols that describe how to use topical anesthetics and systemic analgesics prior to and after test article administration in order to avoid animal pain and distress. They also identify specific clinical signs and lesions that can be used as humane endpoints to allow the investigator to end a study early in order to alleviate animal pain and distress.


Report of an ISRTP Workshop: Progress and barriers to incorporating alternative toxicological methods in the US (PDF | 290 KB)

International Society of Regulatory Toxicology and Pharmacology.

The objective of this workshop [was] to explore progress to date in implementing new, revised and alternative toxicological test methods across regulatory evaluation frameworks and decision-making programs in the U.S., identify barriers to progress and explore potential bridges to facilitate overcoming such barriers. These barriers may be technical, regulatory, economic or societal. Slides and speakers notes for all presentations are also available.


Graphic depicting a human eye and danger sign Evaluation of the Current Validation Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants

The Interagency Coordinating Committee on the Validation of Alternative Methods.

A November 2005 report on the utility of four in vitro ocular toxicity test methods.


Murine Local Lymph Node Assay (PDF | 9.18 MB)

Interagency Coordinating Committee on the Validation of Alternative Methods; National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods.

The Murine Local Lymph Node Assay (LLNA) is an alternative test method for assessing the allergic contact dermatitis potential of chemicals.


Chemicals Testing - Guidelines

Organisation for Economic Co-operation and Development.

Provides internationally agreed testing methods used by government, industry and independent laboratories to characterize potential hazards of new and existing chemical substances.


Progress and Barriers To Incorporating Alternative Toxicological Methods in the U.S.

International Society of Regulatory Toxicology and Pharmacology.

The program and presentation slides from a 2005 workshop that examined progress and barriers to date in implementing new, revised and alternative toxicological test methods across regulatory evaluation frameworks and decision-making programs.


Sens-it-iv Project

An EU-funded research project with the goal of developing alternatives to animal testing when assessing skin or respiratory allergens.


Guinea pig in a PVC environmental enrichment tube Alternative Approaches and Animal Use in the Pharmaceutical Industry (PPT | 3.87 MB)

Europa, Gateway to the European Union.

This presentation offers a view of alternatives and the 3Rs from the research and development perspective of the pharmaceutical industry.


Alternative Methods Publications

Institute for In Vitro Sciences, Inc.

A searchable citation database and links to full-text documents on alternative methods in toxicology.


Refinement, Reduction, and Replacement of Animal Use for Regulatory Testing: Current Best Scientific Practices for the Evaluation of Safety and Potency of Biologicals

ILAR Journal.

This article describes animal testing characteristics and regulatory framework. It also discusses current trends in the development and implementation of alternatives, particularly in the field of biologicals testing (i.e. vaccine development).


Regulatory Testing and Animal Welfare

Institute for Laboratory Animal Research.

This collection of journal articles discusses best practices and animal welfare issue in regards to regulatory testing and toxicology test methods.


Animals in Research

Society of Toxicology.

Provides links to presentations, abstracts, articles, position statements and a downloadable brochure on the importance of animals in the science of toxicology.