ASTM_E_1439_-_12.pdf

Designation:E143912Standard Guide forConducting the Frog Embryo Teratogenesis Assay-Xenopus(FETAX)1This standard is issued under the fixed designation E1439;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.1.Scope1.1 This guide covers procedures for obtaining laboratorydata concerning the developmental toxicity of a test material.The test utilizes embryos of the African clawed frog,Xenopuslaevis and is called FETAX(Frog Embryo TeratogenesisAssay-Xenopus)(1).2Some of these procedures will be usefulfor conducting developmental toxicity tests with other speciesof 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 therequired mortality,malformation,and growth-inhibition dataare described.Special needs or circumstances might requiredifferent types of exposure and data concerning other effects.Some of these modifications are listed in Appendix X2although other modifications might also be necessary.When-ever these procedures are altered or other species used,theresults of tests might not be comparable between modified andunmodified procedures.Any test that is conducted usingmodified procedures should be reported as having deviatedfrom the guide.1.3 These procedures are applicable to all chemicals eitherindividually or in formulations,commercial products or mix-tures that can be measured accurately at the necessary concen-trations in water.With appropriate modification these proce-dures can be used to conduct tests on the effects of temperature,dissolved oxygen,pH,physical agents,and on materials suchas aqueous effluents(see Guide E1192),surface and groundwaters,leachates,aqueous and solid phase extracts,and solidphase 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 thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.1.5 This guide is arranged as follows:SectionReferenced Documents2Terminology3Summary of Guide4Significance and Use5Safety Precautions6Apparatus7Water for Culturing Xenopus adults8Requirements8.1Source8.2Treatment8.3Characterization8.4FETAX Solution Water9Requirements9.1Formulation9.2Test Material10General10.1Stock Solution10.2Test Organisms11Species11.1Source11.2Adults11.3Breeding11.4Embryos11.5Procedure12Experimental Design12.1Temperature and pH Requirements12.2Beginning the Test12.3Renewal12.4Duration of Test12.5Exogenous Metabolic Activation System(MAS)12.6Biological Data12.7Analytical Methodology13Acceptability of the Test14Documentation15Keywords16Appendixes17X1.List of Alternative SpeciesAppendix X1X2.Additional Endpoints and Alternative ExposuresAppendix X2X3.Concentration Steps for Range-Finding TestsAppendix X3X4.Microsome Isolation Reagents and NADPH GeneratingSystem Components,Appendix X4References1This guide is under the jurisdiction ofASTM Committee E50 on EnvironmentalAssessment,Risk Management and Corrective Action and is the direct responsibil-ity of Subcommittee E50.47 on Biological Effects and Environmental Fate.Astandard guide is a document,developed using the consensus mechanisms ofASTM,that provides guidance for the selection of procedures to accomplish aspecific test but which does not stipulate specific procedures.Current edition approved Dec.1,2012 Published January 2013.Originallyapproved in 1991.Last previous edition approved in 2004 as E1439 98(2004).DOI:10.1520/E1439-12.2The boldface numbers in parentheses refer to the list of references at the end ofthe text.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 2.Referenced Documents2.1 ASTM Standards:3D1193 Specification for Reagent WaterE729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes,Macroinvertebrates,and Amphib-iansE943 Terminology Relating to Biological Effects and Envi-ronmental FateE1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE1192 Guide for Conducting Acute Toxicity Tests on Aque-ous Ambient Samples and Effluents with Fishes,Macroinvertebrates,and AmphibiansE1391 Guide for Collection,Storage,Characterization,andManipulation of Sediments for Toxicological Testing andfor Selection of Samplers Used to Collect Benthic Inver-tebratesE1525 Guide for Designing Biological Tests with SedimentsE1706 Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Freshwater InvertebratesIEEE/ASTM SI 10 American National Standard for Use ofthe International Sys