BS 6068-2 69-2000 Water quality Physical chemical and biochemical methods Guidelines for selective immunoassays for the determination of plant treatment and pesticide agents《水质 物理 .pdf

1、 | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS 6068-2.69:2000 ISO 150

2、89:2000 ICS 13.060.50 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Water quality Guidelines for selective immunoassays for the determination of plant treatment and pesticide agentsThis British Standard, having been prepared under the direction of the Health and Environment

3、Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 June 2000 BSI 06-2000 ISBN 0 580 34800 8 BS 6068-2.69:2000 Amendments issued since publication Amd. No. Date Comments National foreword This British Standard reproduces verbatim ISO 15089:2000

4、and 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/3/2, Physical, chemical and biochemical methods, which has the responsibility to: aid enquirers to understand the text; present to the r

5、esponsible international/European 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 obtaine

6、d on request to its secretary. 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

7、Glossary Part 2 Physical, chemical 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 suitably qualified persons wit

8、h an apprpriate level of chemical expertise. Standard chemical 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 “Internationa

9、l Standards Correspondence 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

10、 Standard does not of 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 iv, pages 1 to 14, an inside back cover and a back cover. The BSI copyright notice displayed in this document indicates

11、when the document was last issued.Referencenumber ISO15089:2000(E) INTERNATIONAL STANDARD ISO 15089 Firstedition 2000-03-15 WaterqualityGuidelinesforselective immunoassaysforthedeterminationof planttreatmentandpesticideagents Qualit de leau Lignes directrices relatives aux dosages immunologiques sle

12、ctifs pour la dtermination des agents de traitement des plantes et des pesticidesISO15089:2000(E) ii ISO15089:2000(E)iii Contents Page Foreword.iv 1 Scope1 2 Normativereferences1 3 Termsanddefinitions.1 4 Interferences4 5 Principle4 6 Reagents.4 7 Apparatus.5 8 Samplingandsamplepreparation.5 9 Proce

13、dure.6 10 Validitycriteria.7 11 Calculation8 12 Precision.10 13 Testreport10 AnnexA(informative)Flowchartandpipettingscheme.11 AnnexB(informative)Precisiondatafromaninterlaboratorytrial.13 Bibliography14ISO15089:2000(E) iv Foreword ISO(theInternationalOrganizationforStandardization)isaworldwidefeder

14、ationofnationalstandardsbodies(ISO memberbodies).TheworkofpreparingInternationalStandardsisnormallycarriedoutthroughISOtechnical committees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteehasbeenestablishedhas therighttoberepresentedonthatcommittee.Internationalorganizations,governmenta

15、landnon-governmental,in liaisonwithISO,alsotakepartinthework.ISOcollaboratescloselywiththeInternationalElectrotechnical Commission(IEC)onallmattersofelectrotechnicalstandardization. InternationalStandardsaredraftedinaccordancewiththerulesgivenintheISO/IECDirectives,Part3. DraftInternationalStandards

16、adoptedbythetechnicalcommitteesarecirculatedtothememberbodiesforvoting. PublicationasanInternationalStandardrequiresapprovalbyatleast75%ofthememberbodiescastingavote. AttentionisdrawntothepossibilitythatsomeoftheelementsofthisInternationalStandardmaybethesubjectof patentrights.ISOshallnotbeheldrespo

17、nsibleforidentifyinganyorallsuchpatentrights. InternationalStandardISO15089waspreparedbyTechnicalCommitteeISO/TC147, Water quality, SubcommitteeSC2, Physical, chemical, biochemical methods. AnnexesAandBofthisInternationalStandardareforinformationonly.INTERNATIONALSTANDARD ISO15089:2000(E)1 Waterqual

18、ityGuidelinesforselectiveimmunoassaysforthe determinationofplanttreatmentandpesticideagents 1 Scope ThisInternationalStandardspecifiesaguidefortheselectivequantitativeanalysisbyimmunoassaysof environmentalchemicalssuchaspesticides(includinginsecticides)ortheirmetabolitesindrinking,groundand surfacew

19、ater. Theapplicationrangeoftheprocedurefortheanalysisofpesticidesindrinkingwaterappliestomass concentrationsW0,05 g/l.Therefore,thedeterminationlimitshouldbeinthiscaseu0,05 g/l. 2 Normativereferences Thefollowingnormativedocumentscontainprovisionswhich,throughreferenceinthistext,constituteprovisions

20、of thisInternationalStandard.Fordatedreferences,subsequentamendmentsto,orrevisionsof,anyofthese publicationsdonotapply.However,partiestoagreementsbasedonthisInternationalStandardareencouragedto investigatethepossibilityofapplyingthemostrecenteditionsofthenormativedocumentsindicatedbelow.For undatedr

21、eferences,thelatesteditionofthenormativedocumentreferredtoapplies.MembersofISOandIEC maintainregistersofcurrentlyvalidInternationalStandards. ISO5667-1:1980, Water quality Sampling Part 1: Guidance on the design of sampling programmes. ISO5667-2:1991, Water quality Sampling Part 2: Guidance on sampl

22、ing techniques. ISO5667-3:1994, Water quality Sampling Part 3: Guidance on the preservation and handling of samples. ISO/TR13530:1997, Water quality Guide to analytical quality control for water analysis. 3 Termsanddefinitions ForthepurposesofthisInternationalStandard,thefollowingtermsanddefinitions

23、apply. 3.1 affinity strengthofbindingofantibodytoanalyte NOTE Thestrengthisdefinedbytheequilibriumconstant(K)ofthereactionAb+H=AbH,whereAb=antibody combiningsiteandH=hapten;KisgivenbythemassactionequationK=c AbH /(c Ab c H ). 3.2 analyte substancetobedetermined 3.3 antibodies serumproteinsproducedin

24、vertebratesinresponsetoimmunizationandwhichselectivelybindtheantigenor hapten,respectivelyISO15089:2000(E) 2 NOTE1 Monoclonalantibodies(mAb)areuniformpopulationsofantibodieswhichareproducedfromasinglecellcloneof hybridomacells. NOTE2 Polyclonalantibodies(pAb)areamixedpopulationofantibodieswhicharepr

25、oducedbyseveralclonesofplasma cells. NOTE3 Recombinantantibodies(rAb)areproducedusingrecombinanttechniquesestablishedingenetechnology. 3.4 antibodyconjugate antibodycovalentlylinkedtoalabelsuchasanenzymeorafluorochrome 3.5 antigen substancethatstimulatestheproductionofantibodiesandreactswiththem 3.6

26、 antiserum immuneserumobtainedfromthebloodofimmunizedvertebratesafterremovalofcellularcomponentsand coagulationfactors NOTE Itusuallycontainsanumberofdifferentantibodieswhichcanexhibitdifferentaffinitiestotheantigen/hapten. 3.7 coatingconjugate macromoleculeboundtothehapten(alsoknownasahapten-carrie

27、rconjugate),whichisimmobilizedtoasolid phase NOTE Itisusedtobindthoseantibodybindingsiteswhicharenotoccupiedbytheanalyte. 3.8 competitiveimmunoassay testwhichdetectstheproportionofantibodybindingsiteswhichhavebeenoccupiedbythesampleanalyte NOTE Thisisachievedbyaddingatracerwhichbindstotheunoccupieda

28、ntibodybindingsitesandproducesthe measuringsignalafterafurtherreaction. 3.9 cross-reactivity extent,towhichanantibodyoranantiserumreactswithasubstancewhichstructurallydiffersfromtheanalyte NOTE1 Thecross-reactivityofanantibodyoranantiserumwiththissubstanceisdeterminedbycomparingthecalibration curves

29、Thereferencecurveobtainedwiththeanalyteisusedasreferencequantity(=100%cross-reactivity).Thecross- reactivityisusuallydeterminedattheIC 50.Theselectivityofanantibodyoranantiserum,respectively,isinverselyrelatedtothe cross-reactivities.Anantibodyoranantiserum,respectively,candisplaydifferentaffinitie

30、stodifferentsubstances.Withagiven substance,thecross-reactivityofanantiserumcanalsovarywithinthemeasuringrange.Usuallythestructureofthe immunogenessentiallydeterminestheselectivity(theso-calledspecificity)ofanantiserum.Ifcross-reactivitiesareduetoa mixedpopulationofantibodiesasoftenoccursinanantiser

31、um,theundesiredantibodiesmayberemovedbycross-absorption. NOTE2 Allcompounds(presentinrelevantconcentrations)thatexhibitcross-reactivitywillcreatefalsepositiveresults. 3.10 enzymeimmunoassay EIA immunochemicalanalysiswhichdetectstheoccupancyofantibodybindingsitesbytheanalytewiththeaidofa tracer(anenz

32、yme-labelledhaptenoranenzyme-labelledantibody)andconsequentlycanbeusedtodetectthe analyteconcentrationinthemedium NOTE Thedetectionprocedureisbasedonthemeasurementoftheenzymeactivityofthetracerbymeansofsubstrate conversion.ISO15089:2000(E)3 3.11 enzymesubstrate substancewhichisconvertedbytheenzymein

33、toaproductthatcanbedetectedbyameasuringdevice 3.12 excessstandard analyteconcentrationwhich,onceexceeded,producesnofurtherdecreaseinthesignalmeasuredinthe immunoassay 3.13 fluorescenceimmunoassay immunochemicaldetectionprocedurewhichisperformedeitherasanimmunoassaywithfluorescentsubstratesor product

34、s,respectively,orasanimmunoassaywithfluorescence-labelledtracersorantibodies,respectively 3.14 hapten substancewhich,becauseofitssmallmolecularsize,doesnotevoketheproductionofantibodiesunlessitis covalentlyboundtoanimmunogen NOTE Pesticidesareexamplesofhaptens. 3.15 heterogeneousimmunoassay testwhic

35、hrequiresaseparationofsolidphase-boundandunboundtracersinordertodetecttheoccupancyof antibodybindingsitesbytheanalyteandthustheanalyteconcentrationinthesample 3.16 immunoassay quantitativeassaywhichisbasedontheselectiveanalyte/antibodybindingandusestracersforthedetectionof thefreeoroccupiedantibodyb

36、indingsites,respectively 3.17 immunogen substancewhichtriggersanimmuneresponseafterinjectionintoavertebrate 3.18 inhibitionconcentration IC analyteconcentrationwhichreducesthemeasuringsignalofthezerostandard(=100%),inthecaseofIC 50 to 50%ofthezerostandard 3.19 luminescenceimmunoassay LIA immunochemi

37、calassaywhichiseitherperformedasanimmunoassaydetectingluminescentsubstrateorproduct, respectively,orluminescence-labelledtracer 3.20 solidphaseimmunoassay heterogeneousimmunoassaywhichuses(dependingonthetesttype)antibodiesorcoatingconjugates, immobilizedtoasolidphase NOTE Usually,bothtesttypesarekno

38、wnasELISA(enzyme-linkedimmunosorbentassay)whenenzyme-labelledtracers areused. 3.21 tracer labelledhapten(orantigen)orlabelledantibodywhich,inthecaseofcompetitiveimmunoassays,isusedtodetect thepercentageofantibodybindingsitesnotbeingoccupiedbytheanalyteISO15089:2000(E) 4 3.22 zerostandard analyte-fre

39、estandard(methodblank)whichisusedforcalibration 4 Interferences Interferencesarecausedbyimpropersampling,forinstanceduetothechoiceofequipmentormaterialswhich adsorborliberatethesubstancestobeanalysed.Assayconditions,forinstancepHorsamplecomponentssuchas metalions,humicacids,salinityandsolventsinflue

40、ncingthetestcomponents(forinstancematrixeffects),can interferewiththeanalysis.Theinfluenceofmatrixeffectsmaybeassessedbyspikingsampleswithknown amountsoftheanalyte. 5Principle Immunoassaysaremethodswhichuseantibodiesproducedagainstdefinedanalytesoranalytegroupsas biochemicalsensorsforthequantificati

41、onofanalyteconcentrations.Theseassaysareparticularlyusefulas screeningassays.Allimmunoassaysforthedetectionofhaptensarebasedontheprincipleofthecompetitive immunoassay.Assaysforpesticideshavebeenreported(seereferences1to4inthebibliography).Atypical procedure,asanexampleofanEIA,isdescribedbelow. Theso

42、lidphasevariantrequireseitherimmobilizedantibodiesanddissolvedhaptentracers(varianta,direct immunoassay)orimmobilizedcoatingconjugateanddissolvedantibodytracers(variantb,indirectimmunoassay) inconstantratios.Theantibodiesareappliedinlimitingamounts.Thereforecomponentswhicharenotboundto thecoatedsoli

43、dphasecanberemovedbywashingpriortothefinaldetection.Thisalsoincludesmostofthe interferingmatrixeffects.Themoretracermoleculesarebound,theloweristheanalyteconcentrationinthe sample. Theimmobilizationcanbeachievedbypassiveadsorptionorbycovalentbindingtofunctionalgroupsofthesolid phase. Thecalibrationi

44、sperformedwithsolutionsofknownanalyteconcentrations. 6 Reagents 6.1 General Usereagentsandwaterofhighpurity(forinstance“forresidueanalysis“).Informationontypeandoriginof antibodiesorantisera,respectively,aswellasthecross-reactivitiesshallbestatedinthereport.Informationon storageandstabilityoftheused

45、reagentsshallberequestedfromthesupplier.Theselectivityoftheantibodiesor antiserum,respectively,shallguaranteethatsampleconcentrationsdeducedfromacalibrationcurvedonot deviatebymorethan 10%fromtheactualanalyteconcentrationsinthesesamples.Ifthespecificitiesofthe antibodiesorantiserum,respectively,arel

46、ow,theinterferinganalyteshallberemovedfromthesamplesby suitablechemical-physicalprocedures.Cross-reactingantibodiesofanantiserumcanberemovedbycross- absorption. 6.2 Bufferedwashingsolution,usedforwashingforinstancephosphatebufferedsaline(PBS;pH7,6)witha phosphateconcentrationof80mmol/l(preparedwithN

47、aH 2 PO 4 andNa 2 HPO 4 )andasodiumchloride concentrationof85g/l. 6.3 Acid,forinstancesulfuricacid,c(H 2 SO 4 )=1mol/l,foradjustingthepHandstoppingtheenzymereaction. 6.4 Base,forinstancesodiumhydroxide,c(NaOH)=1mol/l,forpHadjustmentandforstoppingtheenzyme reaction.ISO15089:2000(E)5 7 Apparatus 7.1 S

48、olidphase,consistingofplastics,glassormagneticparticlesforuseinheterogeneousimmunoassays,for instancemicrowellplates,testtubes,beads,magneticparticles,ormembranes. 7.2 Multipipettes,forinstancevariablepipettes10 lto500 l,fixedvolumepipettes10 l,100 l,200 l, multichannelpipettesforinstance300 l,anddi

49、spensers. 7.3 Magneticrack,consistingofarackwithamagneticbasetobeusedduringthewashingstepfor immunoassayswithantibodiesboundtoferromagneticparticles. 8 Samplingandsamplepreparation 8.1 Sampling PerformsamplinginaccordancewithISO5667-1,ISO5667-2andISO5667-3.Theimmunoassayisgenerally performedwithrawwatersamples.Samplepretreatmentshouldbeconsidered,ifnecessary.Itispossibleto concentratesamplesbysuitablechemical-physicalprocedures. 8.2 Coatingofsolidphases Coatedsolidphasescanbeobtainedcommercially.Otherwisetheyshouldbepreparedforenzymeimmunoassays a