1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS 6068-5.28:2000 ISO 107
2、06:2000 ICS 13.060.70 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Water quality Determination of long term toxicity of substances to Daphnia magna Straus (Cladocera, Crustacea)This British Standard, having been prepared under the direction of the Health and Environment Sec
3、tor Committee, was published under the authority of the Standards Committee and comes into effect on 15 May 2000 BSI 05-2000 ISBN 0 580 34842 3 BS 6068-5.28:2000 Amendments issued since publication Amd. No. Date Comments National foreword This British Standard reproduces verbatim ISO 10706:2000 and
4、implements it as the UK national standard. The UK participation in its preparation was entrusted by Technical Committee EH/3, Water quality, to Subcommittee EH/3/5, Biological methods, which has the responsibility to: aid enquirers to understand the text; present to the responsible international/Eur
5、opean committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. A list of organizations represented on this subcommittee can be obtained on request to its secretar
6、y. This is one of a series of standards on water quality, others of which have been, or will be, published as Sections of BS 6068. The various Sections of BS 6068 are comprised within Parts 1 to 7, which, together with Part 0, are listed below. Part 0 Introduction Part 1 Glossary Part 2 Physical, ch
7、emical and biochemical methods Part 3 Radiological methods Part 4 Microbiological methods Part 5 Biological methods Part 6 Sampling Part 7 Precision and accuracy Note. The tests described in this British Standard should only be carried out by suitable qualified persons with an appropriate level of b
8、iological expertise. Standard biological procedures should be followed throughout. Cross-references The British Standards which implement international publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspond
9、ence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of
10、itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, the ISO title page, pages ii to v, a blank page, pages 1 to 17 and a back cover. The BSI copyright notice displayed in this document indicates when the document was last issu
11、ed.Referencenumber ISO10706:2000(E) INTERNATIONAL STANDARD ISO 10706 Firstedition 2000-04-01 WaterqualityDeterminationoflongterm toxicityofsubstancesto Daphnia magna Straus(Cladocera, Crustacea) Qualit de leau Dtermination de la toxicit long terme de substances vis-vis de Daphnia magna Straus (Clado
12、cera, Crustacea)ISO10706:2000(E) ii ISO10706:2000(E)iii Contents Page Foreword.iv Introduction.v 1 Scope1 2 Normativereferences1 3 Principle1 4 Testenvironment.1 5 Reagentsandmaterials2 6 Apparatus.3 7 Treatmentandpreparationofsamples.3 7.1 Specialprecautionsforsamplingandtransportationofwateroreffl
13、uentsamples3 7.2 Preparationofsolutionsofsubstancestobetested.3 8 Procedure.4 8.1 Controls4 8.2 Selectionofexposureconcentrations4 8.3 Renewalofexposuresolutions5 8.4 Introductionofanimalsintothetestsystem5 8.5 Feedingoforganisms.5 8.6 Observationsandmeasurements5 8.7 Validityofthetest6 9 Dataanalys
14、isandexpressionofresults.6 10 Testreport7 AnnexA(informative)OECDM4andM7mediaPreparationandacclimationofElendtM4andM7 media.9 AnnexB(informative)ASTMreconstitutedhardfreshwater.12 AnnexC(informative)Datacollectionsheet13 AnnexD(informative)Calculationoftime-weightedmeanconcentration15 Bibliography17
15、ISO10706:2000(E) iv Foreword ISO(theInternationalOrganizationforStandardization)isaworldwidefederationofnationalstandardsbodies(ISO memberbodies).TheworkofpreparingInternationalStandardsisnormallycarriedoutthroughISOtechnical committees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteeha
16、sbeenestablishedhas therighttoberepresentedonthatcommittee.Internationalorganizations,governmentalandnon-governmental,in liaisonwithISO,alsotakepartinthework.ISOcollaboratescloselywiththeInternationalElectrotechnical Commission(IEC)onallmattersofelectrotechnicalstandardization. InternationalStandard
17、saredraftedinaccordancewiththerulesgivenintheISO/IECDirectives,Part3. DraftInternationalStandardsadoptedbythetechnicalcommitteesarecirculatedtothememberbodiesforvoting. PublicationasanInternationalStandardrequiresapprovalbyatleast75%ofthememberbodiescastingavote. Attentionisdrawntothepossibilitythat
18、someoftheelementsofthisInternationalStandardmaybethesubjectof patentrights.ISOshallnotbeheldresponsibleforidentifyinganyorallsuchpatentrights. InternationalStandardISO10706waspreparedbyTechnicalCommitteeISO/TC147, Water quality, SubcommitteeSC5, Biological methods. AnnexesA,B,CandDofthisInternationa
19、lStandardareforinformationonly.ISO10706:2000(E)v Introduction ThisInternationalStandarddefinesaprocedureforthedeterminationofthelongtermsublethaltoxicityof chemicals,watersandwastewaterstothewater-flea Daphnia magna Straus. The methodology is adapted from a guidelineproducedbytheOrganisationforEcono
20、micCo-operationandDevelopment(OECDGuideline211,see reference1intheBibliography).INTERNATIONALSTANDARD ISO10706:2000(E)1 WaterqualityDeterminationoflongtermtoxicityofsubstances to Daphnia magna Straus (Cladocera, Crustacea) WARNINGActivatedsludgeandsewagecontainpotentiallypathogenicorganisms.Therefor
21、e, appropriateprecautionsshouldbetakenwhenhandlingthem.Toxictestcompoundsandthosewhose propertiesareunknownshouldbehandledwithcare. 1 Scope ThisInternationalStandarddescribesamethodforthedeterminationofthelongtermsublethaltoxicityto Daphnia magna Straus (Cladocera, Crustacea)o f: a) chemicalsubstanc
22、es,whicharesolubleundertheconditionsofthetest,orcanbemaintainedasstable suspensionsordispersionsundertheconditionsofthetest, b) industrialorsewageeffluents,treatedoruntreated,afterdecanting,filtration,orcentrifugation, c) surfaceorgroundwaters. NOTE ThisInternationalStandardisadaptedfromOECDGuidelin
23、e211 1 . 2 Normativereferences Thefollowingnormativedocumentscontainprovisionswhich,throughreferenceinthistext,constituteprovisionsof thisInternationalStandard.Fordatedreferences,subsequentamendmentsto,orrevisionsof,anyofthese publicationsdonotapply.However,partiestoagreementsbasedonthisInternationa
24、lStandardareencouragedto investigatethepossibilityofapplyingthemostrecenteditionsofthenormativedocumentsindicatedbelow.For undatedreferences,thelatesteditionofthenormativedocumentreferredtoapplies.MembersofISOandIEC maintainregistersofcurrentlyvalidInternationalStandards. ISO5667-2, Water quality Sa
25、mpling Part 2: Guidance on sampling techniques. ISO5667-16, Water quality Sampling Part 16: Guidance on biotesting of samples. 3Principle Daphnia magna females, less than 24 h old, are exposed for a period of 21 days to a test substance, industrial or sewageeffluentorsurface/groundwateraddedtodiluti
26、onwaterinarangeofconcentrationsinasemi-staticor flow-throughsystem.Thesurvivalofparentsisrecordedtogetherwiththenumberofliveoffspringproducedper liveparentattheendofthetest. Thesurvivalisreportedandthereproductiveoutputofexposedparentslivingattheendofthetest(expressedina numberofterms)iscomparedtoth
27、oseofthecontrolparents. 4 Testenvironment Thetestatmosphereshallbefreefromvapoursordustswhichmaybetoxicto Daphnia magna Straus.ISO10706:2000(E) 2 Theexposuresolutionsshallnotbeaerated. Thedissolvedoxygenconcentrationintheexposuresolutionsshallbeabove3mg/landthepHshallbewithinthe rangeofpH6topH9andno
28、tchangebymorethan1,5pHunitsforthedurationofthetest.Hardnessshallbe above140mg/l(asCaCO 3 ),whichhasbeendemonstratedtobenecessarytopromotereproductiveperformance thatshallmeetthevaliditycriteria(see8.7). Thephotoperiodofthetestshallbe16hoflightand8hofdarkness.Theintensityshallbeintherange600luxto 800
29、luxbutnotexceeding1200lux. Thetemperatureofthetestexposuresshallbemaintainedwithintherangeof18Cto22Candthetemperature forthetestshallnotvarybymorethan2C(forinstance18Cto20C,19Cto21Cor20Cto22C) throughoutthetest. 5 Reagentsandmaterials 5.1 Testorganism: Daphnia magna Straus (Cladocera, Crustacea), he
30、reafter referred to as D. magna, obtained byacyclicalparthenogenesisforatleastthreegenerationsunderspecifiedcultureconditions. Theageandthesource(includingclone,ifpossible)ofthe D. magna culture shall be indicated in the test report, sincethesensitivityof D. magna to toxicants can be affected by the
31、 source of the culture. Theanimalsusedforthetestshallbelessthan24holdandbefromthesecondtofifthbrood.The D. magna shall befromahealthystockshowingnosignsofstresssuchasmortality 20%,presenceofmales,ephippia,or discolouredanimalsandthereshallbenodelayintheproductionofthefirstbrood. Thestockanimalsshall
32、bemaintainedincultureconditions(light,temperature,medium,feedingandanimalsper unitvolume)similartothoseinthetest.Ifthecultureconditionsdifferfromtestconditions,itisrecommendedthat onegenerationbeacclimatedunderthetestconditionsforaboutthreeweekssoastoavoidstressingtheparent animals. 5.2 Dilutionwate
33、r,syntheticasdescribedin5.3oruncontaminatednaturalwaterswithcomparablepHand hardnesscharacteristicstothesedilutionwatersforculturingandtesting. 5.3 Syntheticdilutionwater,preparedusingoneofthefollowingmethods: a)OECDM4andM7media(seeinformativeannexA); b)ASTMreconstitutedhardfreshwater(seeinformative
34、annexB). Thedilutionwatershallbeaerateduntilthedissolvedoxygenconcentrationhasreached95%saturationandthe pHhasstabilized.Ifnecessary,adjustthepHto8,0 0,5byaddingsodiumhydroxide(NaOH)solutionor hydrochloricacid(HCl)solution.Thedilutionwaterpreparedinthiswayshallnotbefurtheraeratedbeforeuse. Itisrecom
35、mendedthattheTOClevelsbe 5mg/lbeforetheadditionofalgae.Theuseofdilutionwatercontaining EDTA(forinstanceM4andM7)isnotrecommendedwhentestingcompoundscontainingmetalsbecause chelationmayreducethetoxicityofthecompound. Ifthetesthastobeperformedforpurposesnecessitatingtheuseofadilutionwaterwithcharacteri
36、sticsdiffering fromthosedescribedabove,mentionisnecessaryinthetestreportofthemaincharacteristics(forinstancepH, hardness,TOC,COD)ofthesyntheticdilutionwaterused.ISO10706:2000(E)3 6 Apparatus Ordinarylaboratoryapparatusandinparticularthefollowing. 6.1 Environmentalcontrols,forcontrollingtemperature,p
37、hotoperiodandlightintensity. 6.2 Measuringapparatusand/orinstruments,formeasuringdissolvedoxygen,pH,hardness,totalorganic carbon,chemicaloxygendemand,lightintensityandtemperature. 6.3 Testcontainers,ofchemicallyinertmaterialandofsufficientcapacityforthetests(forexample50mlor 100mlglasstesttubesorbea
38、kers)andwhicharecleananduncontaminated. 7 Treatmentandpreparationofsamples 7.1 Specialprecautionsforsamplingandtransportationofwateroreffluentsamples SamplingofwateroreffluentshallbecarriedoutinaccordancewiththegeneralprocedurespecifiedinISO5667-2. Bottlesshallbecompletelyfilledtoexcludeair. Thepres
39、ervationandstorageofwateroreffluentsamplesshallbecarriedoutinaccordancewithISO5667-16;the followingisonlyasummary.Carryoutthetoxicitytestassoonaspossible,ideallywithin12hofcollection.Ifthis timeintervalcannotbeobserved,coolthesample(0Cto4C)andtestthesamplewithin24h.Iftestingcannot becarriedoutwithin
40、48h,thesamplemaybefrozen(below 18C)fortestingwithin2monthsofcollection. Allportions(subsamples)shallbepretreatedidentically(i.e.iffreezingofunstablewaterisnecessaryallportions, includingthoseneededforthefirstday,shallbefrozenpriortotesting;seeISO5667-16). Becauseofthedurationofthistest(21days)andthe
41、periodicrenewalofsolutions,asufficientnumberofportions ofthesamplemustbefrozentorenewtestsolutionsandtorepeatthetest,ifneeded(reservesamples).The minimumvolumeofthefrozenportionsisdependentuponthetoxicityofthesampletobetested.Thevolumeof sampleneededforasemi-statictestwithtendaphnidsexposedindividua
42、llyin100mlofundilutedsampleis9l; additionalsamplewillberequiredfordilutedexposures. Ifthetestisconductedonsiteorclosetothesamplingsite,freshsamplesmaybeusedtoreplacethetest solutions.Inthiscasethevariationsatthesamplingsiteareincorporatedintothetestdesign. 7.2 Preparationofsolutionsofsubstancestobet
43、ested 7.2.1 Preparationofstocksolutions Astocksolutionofthesubstance(s)tobetestedshallbepreparedbydissolvingaknownquantityofthe substance(s)inaspecifiedvolumeofdilutionwater,deionizedwaterordistilledwaterinaglasscontainer.The stocksolutionshallbepreparedimmediatelybeforepreparingtheexposuresolutions
44、unlessthesubstance(s) is/areknowntobestableindefinedstorageconditions,inwhichcasethestocksolutionmaybepreparedin advanceoftestingandstoredintheseconditions. Stocksolutionsorsuspensionsofsubstance(s)whicharepoorlysolubleinwatercanbesolubilizedordispersed directlyinthemediumwithultrasonicdispersion,an
45、d/orstirringorwithsolventsordispersantsoflowtoxicityto D. magna as discussed in ISO 5667-16. If a solvent is used, the concentration of the solvent in the stock solution shallbesuchthattheconcentrationinthehighestexposuresolutiondoesnotexceed0,1ml/l. Theuseoforganicsolventsshouldbeavoided.Iftheyarer
46、equiredorganicsolventssuchasacetone,ethanol, methanol,dimethylformamide,triethyleneglycolordispersantssuchasCremophorRH40,methylcellulose0,1%, andHCO-40maybeusedtoproduceasuitablyconcentratedstocksolution.Theyarenottoxicto D. magna at 0,1ml/lconcentrations.Nosingleprocedureforthepreparationofstockso
47、lutionsofpoorlysolublesubstancescan berecommendedduetothedifferingnatureofchemicals.ISO10706:2000(E) 4 7.2.2 Preparationofexposuresolutions Theexposuresolutionsshallbeprepared(8.2)byaddingthestocksolutions(7.2.1)oreffluentsamples(7.1)tothe dilutionwater(5.2)inspecifiedquantities(see8.2). Ifthestocks
48、olutionsarepreparedindeionizedordistilledwater,nomorethan100mlofstocksolutionshallbe addedtoeachlitreofdilutionwater. Thechosenconcentrationscanalsobepreparedseparatelybythedirectadditionofthetestsubstancetodilution waterwheretheamountstobeaddedcanbeaccuratelyweighedoutorpipettedforliquids. Ifthesam
49、plepHisnotbetween6and9,thetestshallbecarriedoutafteradjustingthepHtothevalueofthe nearestlimit(6or9)usingsolutionsofhydrochloricacidorsodiumhydroxide. NOTE Testingwatersamplesatconcentrationsabove100ml/lmayreducethereproductionandsurvivalof D. magna becauseofdeficiencyinthemedium(forinstancehardness).Identifyingeffectsofsuchdeficienciesmayrequiretheadditionof thesamesaltstothesampleasinthedilutionwater. 8 Procedure 8.1 Controls Everytestshallincludeacontrolcontainingnotestsubstance. Twocontrolsarerequiredwhenasolventordispersantisused.Onecont
