1、Designation: E1022 94 (Reapproved 2013)Standard Guide forConducting Bioconcentration Tests with Fishes andSaltwater Bivalve Mollusks1This standard is issued under the fixed designation E1022; the number immediately following the designation indicates the year oforiginal adoption or, in the case of r
2、evision, 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 describes procedures for obtaining laboratorydata concerning bioconcentration of a tes
3、t 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 Oth
4、er 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 othertes
5、ts. 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 nece
6、ssary 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 (s
7、ee 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 te
8、rms ofwhole body for fishes and in terms of total soft tissue forbivalve mollusks. For fishes and scallops consumed byhumans, 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 organo-
9、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 6.4Cl
10、eaning 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 Treatment 1
11、0.6Holding 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 Test
12、 13Calculation of Results 14Documentation 15Keywords 161This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fateand is the direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved March 1, 2013. Published
13、March 2013. Originallyapproved in 1984. Last previous edition approved in 2007 as E1022 94(2007).DOI: 10.1520/E1022-94R13.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States11.6 The values stated in SI units are to be regarded asstandard.
14、 No other units of measurement are included in thisstandard.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-b
15、ility of regulatory limitations prior to use. Specific precau-tionary statements are given in Section 7.2. Referenced Documents2.1 ASTM Standards:2D1129 Terminology Relating to WaterE729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates, and Amphib-iansE943 T
16、erminology Relating to Biological Effects and Envi-ronmental FateE1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE1191 Guide for Conducting Life-Cycle Toxicity Tests withSaltwater MysidsE1193 Guide for Conducting Daphnia magna Life-CycleToxicity TestsE1241 Guide
17、for Conducting Early Life-Stage Toxicity Testswith FishesE1295 Guide for Conducting Three-Brood, Renewal Toxic-ity 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,
18、” “should,” “may,” “can,” 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 c
19、onnection with factors that 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
20、questionable. Terms such as “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
21、” is never used as a synonym for either “may” or “can.”3.1.2 For definitions of other terms used in this guide, referto Terminologies D1129 and E943 and Guide E729. 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 appa
22、rent steady-state bioconcentration factora BCFthat does not change significantly over a period of two to fourdays at a uniform concentration (as defined in 11.10.3.2)ofthetest material in the solution containing the organism, that is, theBCF that exists when uptake and depuration are equal andbiocon
23、centration (net accumulation) is zero for two to fourdays.3.2.2 bioaccumulationthe net accumulation of a substanceby an organism as a result of uptake from all environmentalsources.3.2.3 bioconcentrationthe net accumulation of a sub-stance by an aquatic organism as a result of uptake directlyfrom aq
24、ueous solution.3.2.4 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 sam
25、e material in thesolution 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 depuration
26、loss of a substance 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
27、 a bioconcentrationtest 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
28、 when placed indilution water to which no test material has been added, usuallyexpressed in units of reciprocal time.3.2.9 effective average exposure concentrationthe averageconcentration, at any time during the uptake phase of abioconcentration test, of test material in the test solution duringthe
29、preceding period 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 oftes
30、t material is constant.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 uptakeacquisit
31、ion of a substance from the environ-ment by an organism as a result of any active or passiveprocess.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Do
32、cument Summary page onthe ASTM website.E1022 94 (2013)23.2.12 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 bioconcentration testduring which
33、organisms are exposed to test material intention-ally added to dilution water. (Although uptake and depurationboth 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 ex
34、ceeds uptake during 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
35、. Summary of Guide4.1 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 mat
36、erial has beenadded, 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 t
37、o dilution water, 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, repr
38、esentative organisms 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 wate
39、r samples. If it is desired todetermine whether BCFs and rate constants are dependent onthe concentration of test material in water, additionaltreatments, utilizing different concentrations of test materialduring the uptake phase, must be used.5. Significance and Use5.1 A bioconcentration test is co
40、nducted to obtain 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 s
41、teady-state 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
42、 steady-state 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 unti
43、linfinite time.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 du
44、ring tests.5.5 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, particulat
45、e matteris deliberately 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 o
46、f the testsubstance 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
47、per se forstable 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
48、been reported 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 bioconcentr
49、ation test may 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 E1023)orderiving water-quality criteria because consumer animalsmight be adversely affect
