1、BRITISH STANDARD BS EN 14185-1:2003 Non-fatty food Determination of N-methylcarbamate residues Part 1: HPLC-method with SPE clean-up The European Standard EN 14185-1:2003 has the status of a British Standard ICS 67.060; 67.080.01 BS EN 14185-1:2003 This British Standard was published under the autho
2、rity of the Standards Policy and Strategy Committee on 6 May 2003 BSI 6 May 2003 ISBN 0 580 41780 8 National foreword This British Standard is the official English language version of EN 14185-1:2003. The UK participation in its preparation was entrusted to Technical Committee AW/-/3, Horizontal ana
3、lysis, which has the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Catalogue unde
4、r the section entitled “International Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online. This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct appl
5、ication. Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; m
6、onitor related international and European developments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the EN title page, pages 2 to 16, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates wh
7、en the document was last issued. Amendments issued since publication Amd. No. Date CommentsEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM EN141851 April2003 ICS67.060;67.080.01 Englishversion NonfattyfoodDeterminationofNmethylcarbamateresidues Part1:HPLCmethodwithSPEcleanup AlimentsnongrasDosagesdes
8、rsidusdeN mthylcarbamatesPartie1:MthodeparCLHPavec purificationSPE FettarmeLebensmittelBestimmungvonN MethylcarbamatRckstndenTeil1:HPLCVerfahrenmit ReinigungdurchFestphasenextraktion ThisEuropeanStandardwasapprovedbyCENon20February2003. CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulations
9、whichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatelistsandbibliographicalreferencesconcernings uchnational standardsmaybeobtainedonapplicationtotheManagementCentreortoanyCENmember. ThisEuropeanStandardexistsinthreeofficialversions(Engl
10、ish,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheManagementCentrehasthesamestatusasthe official versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,CzechRepublic,Denmark,Finland,France,Germany,Greece, H
11、ungary,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal,Slovakia,Spain,Sweden,SwitzerlandandUn ited Kingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISATION EUROPISCHESKOMITEEFRNORMUNG ManagementCentre:ruedeStassart,36B1050Brussels 2003CEN Allrightsofexploitationina
12、nyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.EN141851:2003EEN141851:2003(E) 2 Contents page Foreword3 1 Scope 3 2 Normativereferences 3 3 Principle3 4 Reagents.4 5 Apparatus .5 6 Sampling.6 7 Preparationofthesamples .6 8 Procedure .7 9 Calculationoftheresults 8 10 Confirmatoryt
13、ests . 8 11 Precision,RepeatabilityandReproducibility. .8 12 Testreport 9 AnnexA (informative) PrecisionData 11 AnnexB (informative) AlternativeHPLCoperatingconditions .15 Bibliography 16EN141851:2003(E) 3 Foreword Thisdocument(EN141851:2003)hasbeenpreparedbyTechnicalCommitteeCEN/TC275“Foodanalysis
14、Horizontalmethods“,thesecretariatofwhichisheldbyDIN. ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublicationofanidenticaltextor byendorsement,atthelatestbyOctober2003,andconflictingnationalstandardsshallbewithdrawnatthelatest byOctober2003. AnnexesAandBareinformative. Accord
15、ingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizationsofthefollowing countriesareboundtoimplementthisEuropeanStandard:Austria,Belgium,CzechRepublic,Denmark,Finland, France,Germany,Greece,Hungary,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal, Slovakia,Spain,Swed
16、en,SwitzerlandandtheUnitedKingdom. 1Scope ThisEuropeanStandardspecifiesahighperformanceliquidchromatographic(HPLC)methodforthedetermination ofNmethylcarbamatepesticidesincereals,fruitsandvegetables. Themethodhasbeenvalidatedbycollaborativestudyforcarbaryl,carbofuran,methiocarb,methomyl,oxamyland pro
17、poxurparentcompoundsandformethiocarbsulfoxideingreenpeppersandapplesatlevelsbetween 0,08mg/kgand0,9mg/kg. Nocollaborativedataareavailablefortheperformanceofthemethodinthedeterminationofothersignificant metabolitesalthoughitisknownthatthemethodwillnotworkforoxamylandmethomyloximes. 2 Normativereferen
18、ces ThisEuropeanStandardincorporates,bydatedorundatedreference,provisionsfromotherpublications.These normativereferencesarecitedattheappropriateplacesinthetext,andthepublicationsarelistedhereafter.For datedreferences,subsequentamendmentstoorrevisionsofanyofthesepublicationsapplytothisEuropean Standa
19、rdonlywhenincorporatedinitbyamendmentorrevision.Forundatedreferencesthelatesteditionofthe publicationreferredtoapplies(includingamendments). ENISO3696, WaterforanalyticallaboratoryuseSpecificationandtestmethods(ISO3696:1987). 3Principle Thesampleishomogenizedwithacetone,dichloromethaneandlightpetrol
20、eumandthehomogenateiscentrifuged toyieldtwolayersofthesupernatant.Analiquotportionoftheupperlayerisevaporatedtodryness.Optionally,this extractmaybecleanedupbysolidphaseextraction(SPE)usingacartridgepackedwithaminopropylbonded silica.Intheextractsolution,theNmethylcarbamatesaredeterminedbyreversedpha
21、sehighperformanceliquid chromatography(HPLC)withpostcolumnhydrolysis.Themethylamineformedisallowedtoreactwitho phthaldialdehydeand2mercaptoethanolandthederivativesaredetectedwithafluorescencedetector.Forfurther informationonthismethod,see1to4.EN141851:2003(E) 4 4Reagents 4.1General Unlessotherwisesp
22、ecified,usereagentsofrecognizedanalyticalgrade,preferablyforHPLCandpesticideresidue analysis,anddistilledwaterforcleaningofglasswareorwaterofatleastgrade1asdefinedinENISO3696. Labelallstandardcontainerswithnameandpurityofthepesticides.Forthefullchemicalnamesandstructures, seeISO1750. 4.2Safetyaspect
23、sassociatedwithreagents WARNINGManypesticidesaretoxicbyvariousroutesofexposure,especiallyinconcentratedform. Whenworkingwiththesepesticidesconsultsafetydatasheetsofthemanufacturerforinformation. Vapoursfromsomevolatilesolventsaretoxic.Severalofthesesolventsarereadilyabsorbedthroughskin.Usean effecti
24、vefumehoodtoremovevapoursofthesesolventsastheyaresetfree. 4.3 Acetone 4.4 Aceticacid 4.5 Dichloromethane 4.6 Lightpetroleum, boilingrange40Cto60C 4.7 Methanol 4.8 Acetonitrile 4.9 Solventmixture Acetonitrile(4.8)/aceticacid(4.4)99,9+0,1( V/V) 4.10 Water,purifiedbyaLCgradewaterpurificationsystem 4.11
25、 Sodiumacetate 4.12 oPhthaldialdehyde 4.13 Sodiumtetraborate, anhydrous 4.14 2Mercaptoethanol 4.15 Trimethacarb 4.16 MobilephaseA Acetonitrile(4.8)/water(4.10)20+80( V/V),containing2,5mmol/lsodiumacetate(4.11).Priortouse,filterthe mobilephaseAwithgentlesuctionthroughamembranefilter(5.10). 4.17 Mobil
26、ephaseB Methanol(4.7)/water(4.10)20+80( V/V),containing2,5mmol/lsodiumacetate(4.11).Priortouse,filterthe mobilephaseBwithgentlesuctionthroughamembranefilter(5.10).EN141851:2003(E) 5 4.18 MobilephaseC Acetonitrile(4.8)/water(4.10)60+40( V/V),containing2,5mmol/lsodiumacetate(4.11).Priortouse,filterthe
27、 mobilephaseCwithgentlesuctionthroughamembranefilter(5.10). 4.19 OPAreagent Ina1000mlvolumetricflask,dissolve3,8gofsodiumtetraborate(4.13)inapproximately900mlofwater(4.10). Addasolutionof250 mgofophthaldialdehyde(4.12)in10 mlofacetonitrile(4.8).Next,add0,1 mlof 2mercaptoethanol(4.14)anddiluteto1000m
28、lwithwater. Thesolutionisstableforapproximatelyoneweek. 4.20 SPEelutingmixture (optional) Dichloromethane(4.5)/methanol(4.7)99+1( V/V). 4.21 Internalstandardsolution, r =0,05g/ml. Dissolve10mgoftrimethacarb(4.15)in10mlofacetonitrile(4.8)anddilute100lofthissolutionto100mlina volumetricflaskwithsolven
29、tmixture(4.9)togivedilutionA(1g/ml).Inasecondvolumetricflask,dilute5mlof dilutionAto100mlwithacetonitrile(4.8)/water(4.10)20+80( V/V). 4.22 Standardmaterials Nmethylcarbamatepesticidessuchasaldicarb,bendiocarb,bufencarb,butocarboxim,carbanolate,carbaryl, carbofuran,cloethocarb,dioxacarb,dithiocarb,e
30、thidimuron,ethiofencarb,fenobucarb,isoprocarb,methiocarb, methomyl,oxamyl,promecarb,propoxur,thiofanox,themetabolite3hydroxycarbofuranandthesulfoxideand sulfonemetabolitesofaldicarb,butocarboxim,ethiofencarb,methiocarbandthiofanox. 4.23 Pesticidestocksolutions, r =0,05g/ml. Dissolve10mgofastandardma
31、terial(4.22)in10mlofacetonitrile(4.8)anddilute100lofthissolutionto100ml inavolumetricflaskwithsolventmixture(4.9)togivedilutionA(1g/ml).Inasecondvolumetricflask,dilute5ml ofdilutionAto100mlwithacetonitrile(4.8)/water(4.10)20+80( V/V). 4.24 Pesticidestandardsolutions Prepareappropriatestandardsolutio
32、nsbydilutingsuitableamountsofpesticidestocksolutions(4.23)withinternal standardsolution(4.21)/water(4.10)20+80( V/V). 4.25 Keepersolution Mix2gofethyleneglycolwith8mlofacetone(4.3). 5Apparatus 5.1General Glasswareshallbethoroughlycleaned.Hotdetergentsolutionmaybeusedforcleaning,butafterwardsthe glas
33、swareshallbewellrinsedwithdistilledwaterandacetonebeforedrying. Usuallaboratoryapparatusisusedand,inparticular,thefollowing. 5.2 Chopper 5.3 HomogenizerorhighspeedblenderEN141851:2003(E) 6 5.4 Centrifuge,providedwithpolytetrafluoroethylenetubesofcapacity200ml,andcapableofproducinga rotationalspeedof
34、atleast4000min 1 . 5.5 Waterbath, capableofbeingmaintainedat50Cor60C 5.6 SPEcartridges,packedwith100mgaminopropylbondedsilica,particlesize40m(e.g.BondElut 1) ) (optional) 5.7 DeviceforelutingSPEcartridges (5.6)withsuction(optional) NOTE ApparatusforautomatedSPEelutioniscommerciallyavailable4. 5.8 Hi
35、ghperformanceliquidchromatograph ,equippedwith 5.8.1 Ternarypumpingsystemwithsixportinjectionvalvewitha100lsampleloop,apostcolumnsystem (consistingofacolumnreactor,alowdeadvolumeTpieceandapulsefreereagentpump),afluorescence detectorandaquantificationunitwithanintegratingsystem. 5.8.2 HPLCguardcolumn
36、,stainlesssteelcartridge,10mmlong,4,0mminnerdiameter(e.g.LiChroCART 1) , packedwithSuperspher 1) )60RP8(particlesize4m). 5.8.3 HPLCanalyticalcolumn,stainlesssteelcartridge,250 mmlong,4,0 mminnerdiameter(e.g. LiChroCART 1) ),packedwithSuperspher 1) 60RP8(particlesize4m). 5.8.4 Postcolumnhydrolysiscol
37、umn,stainlesssteel,50 mmlong,4,0 mminnerdiameter,packedwith Aminex 1) A27(15m). 5.9 Ultrasonicbath 5.10 Membranefilters, poresize0,45m 6Sampling Preparethelaboratorysampleaccordingtoagenerallyrecommendedmethodofsamplingtoachievea representativepartoftheproducttobeanalysed. 7 Preparationofthesamples
38、Wherepossible,carryouttheanalysisofsamplesimmediatelyontheirarrivalinthelaboratory.Donotanalysea laboratorysamplewhichiswhollyorextensivelyspoiled. Foranalysistakeonlytheportionofthelaboratorysampletowhichthemaximumresiduelimitapplies.Nofurther plantpartsmayberemoved.Arecordoftheplantpartswhichhaveb
39、eenremovedshallbekept.Thesamplethus preparedisthetestsample. Ifthesamplecannotbeanalysedimmediately,storeitat0Cto5Cfornolongerthan3daysbeforeanalysis. 1) BondElut isatradenameofaproductsuppliedbyAnalytichemInternational,HarborCity,CA,USA.LiChroCART and Superspher aretradenamesofproductssuppliedbyMer
40、ck,Darmstadt,Germany.Aminex isatradenameofaproduct suppliedbyBioRad,Hercules,CA,USA.TheseinformationsaregivenfortheconvenienceofusersofthisEuropeanStandard anddonotconstituteanendorsementbyCENoftheproductsnamed.Equivalentproductsmaybeusediftheycanbeshownto leadtothesameresults.EN141851:2003(E) 7 The
41、reductionofthetestsampleshallbecarriedoutinsuchawaythatrepresentativeportionsareobtained(e.g. bydivisionintofourandselectionofoppositequadrants).Whenthesamplesareinsmallunits(e.g.smallfruits, legumes,cereals),thetestsampleshallbethoroughlymixedbeforeweighingoutthetestportion.Whenthe samplesareinlarg
42、erunits,takewedgeshapedsections(e.g.largefruitsandvegetables)orcrosssections(e.g. cucumbers)whichincludetheoutersurfacefromeachunit. Fromeachtestsample,removethosepartswhichwouldinterferewiththeanalyticalprocedure.Inthecaseof stonefruits,thestonesmayberemoved.Careshallbetakenthataslittleaspossibleof
43、theremaindersuchas juiceorfleshislost.Thebasisforthecalculationoftheresiduemassfractionisthemassoftheoriginaltestsample (withstones). Ifsampleshavetobestoredformorethan3days,theyshallbedeepfrozenat18Corlower.Toensurethat evenafterthawingrepresentativesamplescanbetaken,prepareportionsoftheproductwhichareeachsufficient foroneanalysis. Chopthetestsampleandweighouttestportionsofmassesof15gtoanaccuracyof1%. 8
