ASTM_E_1853M_-_98.pdf

Designation:E 1853M 98Standard Guide forMeasuring the Presence of Planar Organic CompoundsWhich Induce CYP1A,Using Reporter Gene Test Systems1This standard is issued under the fixed designation E 1853M;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(e)indicates an editorial change since the last revision or reapproval.1.Scope1.1 This guide covers the recommended guidelines forperforming a test for presence of organic compounds that bindto the Ah Receptor and induce the CYP1A locus on thevertebrate chromosome.Under appropriate test conditions,induction of CYP1A is evidence that the cells have beenexposed to one or more of these xenobiotic organic com-pounds,including dioxins,furans,coplanar PCBs,and severalpolycyclic aromatic hydrocarbons(PAHs).Detection of induc-tion has been made simple and rapid by the stable integrationof the firefly plasmid such that Ah-receptor binding results inthe production of luciferase.Luciferase production is a func-tion of both the potency of the compound(s)and the concen-tration.This type of Reporter Gene System(RGS)has shownconcentration-response relationships using 2,3,7,8-TCDD,5coplanar PCBs,and several polycyclic aromatic hydrocarbons(PAHs)(1,2).2This guide describes test conditions underwhich solvent extracts of environmental samples(water,tissue,soil,or sediments)may be tested for the presence of CYP1A-inducing organic compounds.1.2 The test procedures presented in this guide have beenpublished previously(1-3).These references should be con-sulted to obtain details regarding the construction and mainte-nance of the cell line,and the response of the cells to variousorganic substances.1.3 All laboratory health and safety procedures should befollowed.This includes the use of glasses,gloves,and otherprotective clothing,when handling the reagents.Informationon toxicity,handling procedures and waste procedures shouldbe reviewed prior to use of all chemicals.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 Documents2Terminology3Application4Summary of Guide5Significance and Use6Interferences7Apparatus8Sample Extraction9Volume Selection9.2Controls9.3Reference Toxicants9.4Treatment of Data10Interpretation of Data10.1Acceptability of Data10.2Reporting Data11Keywords12AnnexP450 Reporter Gene System,Using 101LCellsA1ProcedureA1.1OverviewA1.1.1Culture MaintenanceA1.1.2Instrument CalibrationA1.1.3Testing ProtocolA1.1.4SensitivityA1.2References2.Referenced Documents2.1 ASTM Standards:D 3976 Practice for Preparation of Sediment Samples forChemical Analysis3E 1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their Uses4E 1391 Guide for Collection,Storage,Characterization,andManipulation of Sediments for Toxicological Testing43.Terminology3.1 Definitions:3.1.1 The words“must,”“should,”“may,”“can,”and“might”have very specific meanings in this guide.“Must”isused to express the strongest possible recommendation,justshort of an absolute requirement.“Must”is only used inconnection with factors that relate directly to the acceptabilityof the test.“Should”is used to state that the specific conditionis recommended and ought to be met if possible.Althoughviolation of one“should”is rarely a serious matter,theviolation of several will often render the results questionable.1This guide is under the jurisdiction of ASTM Committee E-47 on BiologicalEffects and Environmental Fateand is the direct responsibility of SubcommitteeE47.09on Biomarkers.Current edition approved March 10,1998.Published September 1998.Originallypublished as E 1853-96.Last previous edition E 1853-96.2The boldface numbers in parentheses refer to a list of references at the end ofthis guide.3Annual Book of ASTM Standards,Vol 11.02.4Annual Book of ASTM Standards,Vol 11.05.1AMERICAN SOCIETY FOR TESTING AND MATERIALS100 Barr Harbor Dr.,West Conshohocken,PA 19428Reprinted from the Annual Book of ASTM Standards.Copyright ASTMTerms such as“is desirable,”“is often desirable,”and“mightbe desirable”are used in connection with less importantfactors.“May”is used to mean“is(are)allowed to,”“can”isused to mean“is(are)able to,”and“might”is used to mean“could possibly.”Thus the classic distinction between“may”and“can”is preserved,and“might”is never used as asynonym for either“may”or“can.”3.2 Definitions of Terms Specific to This Standard:3.2.1 AhR-aryl hydrocarbon receptorin the cell cytoplasmthat binds to planar compounds,receptors forming an AhR-ligand complex,which is translocated to the nucleus where thecomplex activates transcription of the CYP1A gene.3.2.2 B(a)P-be