ASTM E 1439:2012

1.1 This guide covers procedures for obtaining laboratory data concerning the developmental toxicity of a test material. The test utilizes embryos of the African clawed frog, Xenopus laevis and is called FETAX (Frog Embryo Teratogenesis Assay- Xenopus ) ( 1 ) . 2 Some of these procedures will be useful for conducting developmental toxicity tests with other species of frogs although numerous modifications might be necessary. A list of alternative anurans is presented in Appendix X1 . 1.2 A renewal exposure regimen and the collection of the required mortality, malformation, and growth-inhibition data are described. Special needs or circumstances might require different types of exposure and data concerning other effects. Some of these modifications are listed in Appendix X2 although other modifications might also be necessary. Whenever these procedures are altered or other species used, the results of tests might not be comparable between modified and unmodified procedures. Any test that is conducted using modified procedures should be reported as having deviated from the guide. 1.3 These procedures are applicable to all chemicals either individually or in formulations, commercial products or mixtures that can be measured accurately at the necessary concentrations in water. With appropriate modification these procedures can be used to conduct tests on the effects of temperature, dissolved oxygen, pH, physical agents, and on materials such as aqueous effluents (see Guide E1192 ), surface and ground waters, leachates, aqueous and solid phase extracts, and solid phase samples, such as soils and sediments, particulate matter, sediment, and whole bulk soils and sediment. 1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 1.5 This guide is arranged as follows: Section Referenced Documents 2 Terminology 3 Summary of Guide 4 Significance and Use 5 Safety Precautions 6 Apparatus 7 Water for Culturing Xenopus adults 8 Requirements 8.1 Source 8.2 Treatment 8.3 Characterization 8.4 FETAX Solution Water 9 Requirements 9.1 Formulation 9.2 Test Material 10 General 10.1 Stock Solution 10.2 Test Organisms 11 Species 11.1 Source 11.2 Adults 11.3 Breeding 11.4 Embryos 11.5 Procedure 12 Experimental Design 12.1 Temperature and pH Requirements 12.2 Beginning the Test 12.3 Renewal 12.4 Duration of Test 12.5 Exogenous Metabolic Activation System (MAS) 12.6 Biological Data 12.7 Analytical Methodology 13 Acceptability of the Test 14 Documentation 15 Keywords 16 Appendixes 17 X1. List of Alternative Species Appendix X1 X2. Additional Endpoints and Alternative Exposures Appendix X2 X3. Concentration Steps for Range-Finding Tests Appendix X3 X4. Microsome Isolation Reagents and NADPH Generating System Components, Appendix X4 References