【国外标准】 Standard Guide for Conducting the Frog Embryo Teratogenesis Assay-Xenopus (FETAX)

5.1 FETAX is a rapid test for identifying potential developmental toxicity. Data may be extrapolated to other species including mammals. FETAX might be used to prioritize samples for further tests which use mammals. Validation studies using compounds with known mammalian or human developmental toxicity, or both, suggest that the predictive accuracy will exceed 85 % (2) . When evaluating a test material for mammalian developmental toxicity, FETAX must be used with and without a metabolic activation system (MAS). Use of this exogenous MAS should increase the predictive accuracy of the assay to approximately 95 %. The accuracy rate compares favorably with other currently available “ in vitro teratogenesis screening assays” (3). Any assay employing cells, parts of embryos, or whole embryos other than in vivo mammalian embryos is considered to be an in vitro assay.5.2 It is important to measure developmental toxicity because embryo mortality, malformation, and growth inhibition can often occur at concentrations far less than those required to affect adult organisms.5.3 Because of the sensitivity of embryonic and early life stages, FETAX provides information that might be useful in estimating the chronic toxicity of a test material to aquatic organisms.5.4 Results from FETAX might be useful when deriving water quality criteria for aquatic organisms (4).5.5 FETAX results might be useful for studying structure-activity relationships between test materials and for studying bioavailability.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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.1.5 This guide is arranged as follows:  SectionReferenced Documents 2Terminology 3Summary of Guide 4 5Safety Precautions 6Apparatus 7Water for Culturing Xenopus adults 8 Requirements 8.1 Source 8.2 Treatment 8.3 Characterization 8.4FETAX Solution Water 9 Requirements 9.1 Formulation 9.2Test Material 10 General 10.1 Stock Solution 10.2Test Organisms 11 Species 11.1 Source 11.2 Adults 11.3 Breeding 11.4 Embryos 11.5Procedure 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 12.7Analytical Methodology 13Acceptability of the Test 14Documentation 15Keywords 16Appendixes 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 X4References  1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.