1、Designation: E 1022 94 (Reapproved 2007)Standard Guide forConducting Bioconcentration Tests with Fishes andSaltwater Bivalve Mollusks1This standard is issued under the fixed designation E 1022; the number immediately following the designation indicates the year oforiginal adoption or, in the case of
2、 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 describes procedures for obtaining laboratorydata concerning bioconcentration of a
3、test material added todilution waterbut not to foodby freshwater and saltwaterfishes and saltwater bivalve mollusks using the flow-throughtechnique. These procedures also should be useful for conduct-ing bioconcentration tests with other aquatic species, althoughmodifications might be necessary.1.2
4、Other modifications of these procedures might be justi-fied by special needs or circumstances. Although using appro-priate procedures is more important than following prescribedprocedures, the results of tests conducted using unusual pro-cedures are not likely to be comparable to those of many other
5、tests. The comparison of results obtained using modified andunmodified versions of these procedures might provide usefulinformation concerning new concepts and procedures forconducting bioconcentration tests.1.3 These procedures are applicable to all chemicals thatcan be measured accurately at the n
6、ecessary concentrations inwater and in appropriate tissues. Bioconcentration tests areusually conducted on individual chemicals but can be con-ducted on mixtures if appropriate measurements can be made.Some techniques described in this guide were developed fortests on non-ionizable organic chemicals
7、 (see 11.1.2.1) andmight not apply to ionizable or inorganic chemicals.1.4 Results of bioconcentration tests should usually bereported in terms of apparent steady-state and projected steady-state bioconcentration factors (BCFs) and uptake and depura-tion rate constants. Results should be reported in
8、 terms ofwhole body for fishes and in terms of total soft tissue forbivalve mollusks. For fishes and scallops consumed by hu-mans, some results should also be reported in terms of theedible portion, especially if ingestion of the test material byhumans is a major concern. For tests on organic and or
9、gano-metallic chemicals, the percent lipids of the tissue should bereported.1.5 This guide is arranged as follows:SectionReferenced Documents 2Terminology 3Summary of Guide 4Significance and Use 5Safety Precautions 7Apparatus 6Facilities 6.1Construction Materials 6.2Metering System 6.3Test Chambers
10、6.4Cleaning 6.4.4Acceptability 6.5Dilution Water 8Requirements 8.1Source 8.2Treatment 8.3Characterization 8.4Test Material 9General 9.1Radiolabeled Material 9.2Stock Solution 9.3Test Concentration(s) 9.4Test Organisms 10Species 10.1Size 10.2Source 10.3Care and Handling 10.4Feeding 10.5Disease Treatm
11、ent 10.6Holding 10.7Acclimation 10.8Quality 10.9Procedure 11Experimental Design 11.1Dissolved Oxygen 11.2Temperature 11.3Loading 11.4Beginning the Test 11.5Care of Organisms 11.6Feeding 11.7Cleaning 11.8Biological Data 11.9Measurements on Test Solutions 11.10Analytical Methodology 12Acceptability of
12、 Test 13Calculation of Results 14Documentation 15Keywords 161.6 The values stated in SI units are to be regarded as thestandard. The values given in parentheses are for informationonly.1This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fate and is the
13、direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved Oct. 1, 2007. Published October 2007. Originallyapproved in 1984. Last previous edition approved in 2002 as E 1022 94(2002).1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, Wes
14、t Conshohocken, PA 19428-2959, United States.1.7 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 regula
15、tory limitations prior to use. Specific precau-tionary statements are given in Section 7.2. Referenced Documents2.1 ASTM Standards:2D 1129 Terminology Relating to WaterE 729 Guide for Conducting Acute Toxicity Tests withFishes, Macroinvertebrates, and AmphibiansE 943 Terminology Relating to Biologic
16、al Effects and En-vironmental FateE 1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE 1191 Guide for Conducting Life-Cycle Toxicity Testswith Saltwater MysidsE 1193 Guide for Conducting Daphnia Magna Life-CycleToxicity TestsE 1241 Guide for Conducting Early Life-S
17、tage ToxicityTests with FishesE 1295 Guide for Conducting Three-Brood, Renewal Tox-icity Tests with Ceriodaphnia DubiaIEEE/ASTM SI 10 Standard for Use of the InternationalSystem of Units (SI) (the Modernized Metric System)3. Terminology3.1 Definitions:3.1.1 The words “must,” “should,” “may,” “can,”
18、and“might” have very specific meanings in this guide. “Must” isused to express an absolute requirement, that is, to state that thetest ought to be designed to satisfy the specified condition,unless the purpose of the test requires a different design.“Must” is used only in connection with factors tha
19、t relatedirectly to the acceptability of the test (see 13.1). “Should” isused to state that the specified condition is recommended andought to be met if possible.Although violation of one “should”is rarely a serious matter, violation of several will often renderthe results questionable. Terms such a
20、s “is desirable” are usedin connection with less important factors.“ May” is used tomean “is (are) allowed to,” “can” is used to mean “is (are) ableto,” and “might” is used to mean “could possibly.” Thus theclassic distinction between“ may” and “can” is preserved, and“might” is never used as a synon
21、ym for either “may” or “can.”3.1.2 For definitions of other terms used in this guide, referto Terminologies D 1129 and E 943 and Guide E 729. For anexplanation of units and symbols, refer to Standard IEEE/ASTM SI 10.3.2 Definitions of Terms Specific to This Standard:3.2.1 apparent steady-state bioco
22、ncentration factoraBCF that does not change significantly over a period of two tofour days at a uniform concentration (as defined in 11.10.3.2)of the test material in the solution containing the organism, thatis, the BCF that exists when uptake and depuration are equaland bioconcentration (net accum
23、ulation) is zero for two to fourdays.3.2.2 bioaccumulationthe net accumulation of a sub-stance by an organism as a result of uptake from all environ-mental sources.3.2.3 bioconcentrationthe net accumulation of a sub-stance by an aquatic organism as a result of uptake directlyfrom aqueous solution.3.
24、2.4 bioconcentration factor (BCF)the quotient, at anytime during the uptake phase of a bioconcentration test, of theconcentration of a material in one or more tissues of an aquaticorganism at that time, divided by the effective average expo-sure concentration at that time of the same material in the
25、solution which contains the organism, in units of volume ofsolution per mass of organism. (BCFs are usually calculated sothat the volume of solution, for example, 1 L, is aboutcomparable to the mass of tissue, for example, 1 kg, and theBCF is reported without units.)3.2.5 depurationloss of a substan
26、ce from an organism asa result of any active or passive process.3.2.6 depuration curvethe line obtained by plotting themeasured concentration of a test material in aquatic organismsversus time during the depuration phase of a bioconcentrationtest.3.2.7 depuration phasethe portion of a bioconcentrati
27、ontest after the uptake phase and during which the organisms arein dilution water to which no test material has been added.3.2.8 depuration rate constantthe mathematically derivedvalue(s) that expresses how rapidly test material is eliminatedfrom previously exposed aquatic organisms when placed indi
28、lution water to which no test material has been added, usuallyexpressed in units of reciprocal time.3.2.9 effective average exposure concentrationthe aver-age concentration, at any time during the uptake phase of abioconcentration test, of test material in the test solution duringthe preceding perio
29、d of time equal to the shorter of (a) thelength of the uptake phase to that point and (b) one half thetime to apparent steady-state. Effective exposure concentra-tions cannot be calculated until after the time to apparentsteady-state has been determined, unless the concentration oftest material is c
30、onstant.3.2.10 projected steady-state bioconcentration factoraBCF calculated for infinite time (a) from uptake and depurationrate constants derived using an appropriate compartmentalmodel or (b) by fitting an appropriate equation to dataconcerning BCF versus time.3.2.11 uptakeacquisition of a substa
31、nce from the environ-ment by an organism as a result of any active or passiveprocess.3.2.12 uptake curvethe line obtained by plotting themeasured concentration of test material in aquatic organismsversus time during the uptake phase of a bioconcentration test.3.2.13 uptake phasethe portion of a bioc
32、oncentration testduring which organisms are exposed to test material intention-ally added to dilution water. (Although uptake and depuration2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volum
33、e information, refer to the standards Document Summary page onthe ASTM website.E 1022 94 (2007)2both occur during the uptake phase, uptake always predomi-nates at the beginning, but depuration often becomes nearlyequal to uptake at the end of the uptake phase. Occasionallydepuration exceeds uptake d
34、uring a portion of the uptakephase.)3.2.14 uptake rate constantthe mathematically derivedvalue(s) that express how rapidly test material is accumulatedby aquatic organisms during the uptake phase of a bioconcen-tration test, in units of volume of solution per mass of organismper time.4. Summary of G
35、uide4.1 Each of two groups of test organisms of one species isadministered a treatment, consisting of an uptake phase and adepuration phase, using the flow-through technique. The con-trol treatment, in which organisms are exposed during bothphases to dilution water to which no test material has been
36、added, provides a measure of the acceptability of the test bygiving an indication of the quality of the test organisms and thesuitability of the dilution water, food, test conditions, handlingprocedures, etc. In the other treatment the organisms are (a)exposed during the uptake phase to dilution wat
37、er, to which aselected concentration of test material has been intentionallyadded, at least until either apparent steady-state or 28 days isreached and (b) exposed during the depuration phase todilution water to which no test material has been added. Duringboth phases of the test, representative org
38、anisms and watersamples are removed periodically from each test chamber andanalyzed for test material. Apparent steady-state and projectedsteady-state BCFs and uptake and depuration rate constants areusually calculated from the measured concentrations of testmaterial in tissue and water samples. If
39、it is desired todetermine whether BCFs and rate constants are dependent onthe concentration of test material in water, additional treat-ments, utilizing different concentrations of test material duringthe uptake phase, must be used.5. Significance and Use5.1 A bioconcentration test is conducted to o
40、btain informa-tion concerning the ability of an aquatic species to accumulatea test material directly from water. This guide providesguidance for designing bioconcentration tests on the propertiesof the test material so that each material is tested in acost-effective manner.5.2 Because steady-state
41、is usually approached from the lowside and the definition of apparent steady-state is based on astatistical hypothesis test, the apparent steady-state BCF willusually be lower than the steady-state BCF. With the variationand sample sizes commonly used in bioconcentration tests, theactual steady-stat
42、e BCF will usually be no more than twice theapparent BCF.5.3 When both are determined in the same test, the projectedsteady-state BCF will usually be higher than the apparentsteady-state BCF because the models used to calculate theprojected BCF assume that the BCF steadily increases untilinfinite ti
43、me.5.4 The BCFs and rates and extents of uptake and depura-tion will depend on temperature, water quality, the species andits size, physiological condition, age, and other factors (1).3Although organisms are fed during tests, uptake by means ofsorption onto food is probably negligible during tests.5
44、.5 Results of bioconcentration tests are used to predictconcentrations likely to occur in aquatic organisms in fieldsituations as a result of exposure under comparable conditions,except that mobile organisms might avoid exposure whenpossible. Under the experimental conditions, particulate matteris d
45、eliberately minimized compared to natural water systems.Exposure conditions for the tests may therefore not be compa-rable for an organic chemical that has a high octanol-waterpartition coefficient or for an inorganic chemical that sorbssubstantially onto particulate matter. The amount of the testsu
46、bstance in solution is thereby reduced in both cases, andtherefore the material is less available to many organisms.However, sorption might increase bioaccumulation by aquaticspecies that ingest particulate matter (2), or food may be amore important source of residues in fish than water per se forst
47、able neutral organic chemicals that have a Log Kowbetween4 and 6 (3).5.6 Results of bioconcentration tests can be used to comparethe propensity of different materials to be accumulated. Non-ionizable organic chemicals can also be ranked for bioconcen-tration using correlations that have been reporte
48、d betweensteady-state BCFs and physicalchemical properties, such asthe octanolwater partition coefficient and solubility in water(4). However, when such predictions are impossible, exceedthe demonstrated limits of the correlation, or might be other-wise questionable (1, 5), a bioconcentration test m
49、ay benecessary.5.7 Results of bioconcentration tests can also be used tocompare the abilities of different species to accumulate mate-rials. At steady-state the concentration of a nonionizableorganic chemical in individual organisms, and in varioustissues within an organism, will probably be related to theconcentration of lipids in the organisms and tissues (6).5.8 Results of bioconcentration tests might be an importantconsideration when assessing hazard (see Guide E 1023)orderiving water-quality criteria because consumer animalsmight be adversely affected by ingest
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