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TM_E_1279_
_89_2001
Designation:E 1279 89(Reapproved 2001)Standard Test Method forBiodegradation By a Shake-Flask Die-Away Method1This standard is issued under the fixed designation E 1279;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 test method describes procedures for assessing thebiodegradation of chemicals in natural surface water samples.1.2 This test method provides an opportunity to evaluaterates of biodegradation in the presence and absence of naturalsediment materials.It also may provide limited information onthe abiotic degradation rate,and sorption to sediment andvessel walls.1.3 This test method allows for the development of afirst-order rate constant,based on the disappearance of the testcompound with time,and a second-order rate constant,nor-malized for changes in microbial biomass.1.4 This test method requires a chemical specific analyticalmethod and the concentrations of test substance employed aredependent on the sensitivity of the analytical method.1.5 This test method is designed to be applicable to com-pounds that are not inhibitory to bacteria at the concentrationsused in the test method,which do not rapidly volatilize fromwater,that are soluble at the initial test concentration and thatdo not degrade rapidly by abiotic processes,such as hydrolysis.1.6 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.2.Referenced Documents2.1 ASTM Standards:D 1193 Specification for Reagent Water2D 4129 Test Method for Total and Organic Carbon in Waterby Oxidation and Coulometric Detection3E 895 Practice for Determination of Hydrolysis Rate Con-stants of Organic Chemicals in Aqueous Solutions4E 896 Test Method for Conducting Aqueous Direct Pho-tolysis Tests4E 1194 Test Method for Vapor Pressure4E 1195 Test Method for Determining a Sorption Constant43.Summary of Test Method3.1 The shake-flask die-away biodegradation method issimilar to river water die-away tests described by manyauthors,including Degens et al(1),5Eichelberger and Licht-enberg(2),Saeger and Tucker(3),Paris et al(4),and Cripe etal(5).It differs from most die-away methods by providing foran evaluation of the effects of natural sediments on thetransformation of the test compound and by the use of shakingto ensure a dissolved oxygen supply.Each test compound(substrate)is dissolved in water collected from a field site,withand without added natural sediment and with and withoutsterilization.Initial substrate concentrations typically are rela-tively low(g/L),analytical capabilities permitting.Loss oftest compound with time is followed by an appropriate,chemical-specific analytical technique.Changes in microbialbiomass also may be followed by the use of an appropriatetechnique such as bacterial plate counts.Data obtained duringuse of the test method are used to provide the followinginformation:(a)the abiotic degradation rate in the presenceand absence of sediment and(b)the combined biotic andabiotic degradation rate in the presence and absence ofsediment.4.Significance and Use4.1 Most of the simpler methods used to screen chemicalsfor biodegradation potential employ measurements that are notspecific to the test substance,such as loss of dissolved organiccarbon,evolution of respiratory carbon dioxide,or uptake ofdissolved oxygen.Such methods generally are used to evaluatethe transformation of the test substance to carbon dioxide,water,oxides or mineral salts of other elements,or productsassociated with the normal metabolic processes of microorgan-isms(ultimate biodegradability),or both.These methods re-quire the use of relatively high initial concentrations of the testsubstance,generally 10 mg/L or higher,unless the tests areconducted using14C-radiolabeled test compounds.Biodegra-dation tests measuring14C-CO2evolution,for example,can beconducted using initial concentration of test compound at partsper billion.These tests,however,require specialized equip-ment and the custom preparation of appropriately labeledcompound is often very expensive.1This specification is under the jurisdiction of Committee E47 on BiologicalEffects and Environmental Fate and is the direct responsibility of SubcommitteeE47.06 on Environmental Fate of Chemical Substances.Current edition approved Jan.27,1989.Published March 1989.2Annual Book of ASTM Standards,Vol 11.01.3Annual Book of ASTM Standards,Vol 11.02.4Annual Book of ASTM Standards,Vol 11.05.5The boldface numbers in parentheses refer to the list of references at the end ofthis test method.1Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,W