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    EN 1785-2003 en Foodstuffs - Detection of Irradiated Food Containing Fat - Gas Chromatographic Mass Spectrometric Analysis of 2-Alkylcyclobutanones《食品 经过照射处理含脂肪食品的检测 2烷基环丁酮的气相色谱 质.pdf

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    EN 1785-2003 en Foodstuffs - Detection of Irradiated Food Containing Fat - Gas Chromatographic Mass Spectrometric Analysis of 2-Alkylcyclobutanones《食品 经过照射处理含脂肪食品的检测 2烷基环丁酮的气相色谱 质.pdf

    1、BRITISH STANDARD BS EN 1785:2003 Foodstuffs Detection of irradiated food containing fat Gas chromatographic/ mass spectrometric analysis of 2-alkylcyclobutanones The European Standard EN 1785:2003 has the status of a British Standard ICS 67.050 BS EN 1785:2003 This British Standard, was published un

    2、der the authority of the Standards Policy and Strategy Committee on 12 August 2003 BSI 12 August 2003 ISBN 0 580 42454 5 National foreword This British Standard is the official English language version of EN 1785:2003. It supersedes BS EN 1785:1997 which is withdrawn. The UK participation in its pre

    3、paration was entrusted to Technical Committee AW/-/3, Horizontal analysis, 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

    4、 referred to in this document may be found in the BSI Catalogue under 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

    5、provisions of a contract. Users are responsible for its correct application. 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 interpreta

    6、tion, or proposals for change, and keep the UK interests informed; monitor 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 c

    7、over. The BSI copyright notice displayed in this document indicates when the document was last issued. Amendments issued since publication Amd. No. Date CommentsEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM EN1785 August2003 ICS67.050 SupersedesEN1785:1996 Englishversion FoodstuffsDetectionofirradi

    8、atedfoodcontainingfatGas chromatographic/massspectrometricanalysisof2 alkylcyclobutanones ProduitsalimentairesDtectiondalimentsioniss contenantdeslipidesAnalyseparchromatographieen phasegazeuse/Spectromtriedemassedes2 alkylcyclobutanones LebensmittelNachweisvonbestrahltenfetthaltigen LebensmittelnGa

    9、schromatographisch/ massenspektrometrischeUntersuchungauf2 Alkylcyclobutanone ThisEuropeanStandardwasapprovedbyCENon20June2003. CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Up

    10、todatelistsandbibliographicalreferencesconcernings uchnational standardsmaybeobtainedonapplicationtotheManagementCentreortoanyCENmember. ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownl

    11、anguageandnotifiedtotheManagementCentrehasthesamestatusasthe official versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,CzechRepublic,Denmark,Finland,France,Germany,Greece, Hungary,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal,Slovakia,Spain,Sweden,Switzerlandan

    12、dUn ited Kingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISATION EUROPISCHESKOMITEEFRNORMUNG ManagementCentre:ruedeStassart,36B1050Brussels 2003CEN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.EN1785:2003EEN1785:2003(E) 2 Contents Page

    13、 Foreword3 1 Scope 3 2 Normativereferences 3 3 Principle3 4 Reagents.4 5 Apparatus .5 6 Samplingtechnique .6 7 Procedure .6 8 Evaluation.7 9 Limitations8 10 Validation8 11 Testreport 9 AnnexA (informative) Figures . 11 Bibliography 15EN1785:2003(E) 3 Foreword Thisdocument(EN1785:2003)hasbeenprepared

    14、byTechnicalCommitteeCEN/TC 275,“Foodanalysis Horizontalmethods“,thesecretariatofwhichisheldbyDIN. ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublicationofanidenticaltextor byendorsement,atthelatestbyFebruary2004,andconflictingnationalstandardsshallbewithdrawnatthelatest byF

    15、ebruary2004. ThisdocumentsupersedesEN1785:1996. ThisEuropeanStandardwaselaboratedonthebasisofaprotocolduringaconcertedactionoftheEuropean Commission(DG XII C.5).ExpertsandlaboratoriesfromE.U.andEFTAcountries,contributedjointlytothe developmentofthisprotocol. Thepredecessorofthepresentstandard(EN1785

    16、:1996)hasbeenelaboratedfollowingamandateofthe EuropeanCommission. AnnexAisinformative. AccordingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizationsofthefollowing countriesareboundtoimplementthisEuropeanStandard:Austria,Belgium,CzechRepublic,Denmark,Finland, France,Germany,Greece,Hu

    17、ngary,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal, Slovakia,Spain,Sweden,SwitzerlandandtheUnitedKingdom. 1Scope ThisEuropeanStandardspecifiesamethodfortheidentificationofirradiationtreatmentoffoodcontainingfat.Itis basedonthemassspectrometric(MS)detectionofradiationinduced2alk

    18、ylcyclobutanonesaftergas chromatographic(GC)separation1to3. Themethodhasbeensuccessfullytestedininterlaboratorytrialsonrawchicken,pork,liquidwholeegg,salmon andCamembert4to8. Otherstudiesdemonstratethatthemethodisapplicabletoawiderangeoffoodstuffs9to21. 2 Normativereferences ThisEuropeanStandardinco

    19、rporatesbydatedorundatedreferences,provisionsfromotherpublications.These normativereferencesarecitedattheappropriateplacesinthetextandthepublicationsarelistedhereafter.For datedreferences,subsequentamendmentstoorrevisionsofanyofthesepublicationsapplytothisEuropean Standardonlywhenincorporatedinitbya

    20、mendmentorrevision.Forundatedreferencesthelatesteditionofthe publicationreferredtoapplies(includingamendments). ENISO3696:1995, WaterforanalyticallaboratoryuseSpecificationandtestmethods(ISO3696:1987). 3Principle Duringirradiation,theacyloxygenbondintriglyceridesiscleavedandthisreactionresultsinthef

    21、ormationof 2alkylcyclobutanonescontainingthesamenumberofcarbonatomsastheparentfattyacidandthealkylgroupis locatedinringposition2.Thus,ifthefattyacidcompositionisknown,the2alkylcyclobutanonesformedcanbe predicted. The2alkylcyclobutanoneswhichwereanalysedininterlaboratorystudieswere2dodecylcyclobutano

    22、ne(DCB) and2tetradecylcyclobutanone(TCB)whichareformedfrompalmiticandstearicacid ,respectively,during irradiation.Todate,thereisnoevidencethatthe2alkylcyclobutanonescanbedetectedinunirradiatedfoods4,EN1785:2003(E) 4 9to21.The2alkylcyclobutanonesareextractedusingnhexaneornpentanealongwiththefat.Theex

    23、tractis thenfractionatedusingadsorptionchromatographypriortoseparationusinggaschromatographyanddetection withamassspectrometer.Other2alkycyclobutanones,e.g.2(tetradec5enyl)cyclobutanonederivedfromoleic acid,havealsobeenidentifiedinirradiatedfoodstuffs8,15to20. NOTE Asanalternativeprocedureforextract

    24、ionandpurificationofthe2alkylcyclobutanones,supercriticalfluidextraction (SFE),hasbeensuccessfullyemployed18,21.Argentationchromatographyhasbeenusedeffectivelyforthedetectionof foodsirradiatedatverylowdosesorcontainingirradiatedingredients17.Liquidchromatography(LC)GCMScouplinghas beenusedsuccessful

    25、lyasanalternativeprocedureforpurificationanddetection14.Itshould,however,benotedthatthese alternativeprocedureshavenotbeenvalidatedbyinterlaboratorytrials. 4Reagents 4.1General Allreagentsandmaterialsusedshallbeofrecognizedanalyticalgradethepurityofwhichhastobetested regularlybytheanalysisofblanksam

    26、ples.Watershallbeofatleastgrade3accordingtoENISO3696:1995. 4.2nHexane 1) 4.3Sodiumsulfate, anhydrous 4.4Diethylether 4.5Stockstandardsolutions nhexaneorisooctanemaybeusedtopreparesolutionsof2cyclohexylcyclohexanone(5 g/ml),and2 dodecylcyclobutanone 2) and2tetradecylcyclobutanone 2) (100g/ml).Storeat

    27、20C. 4.6Workingstandardsolutions nhexaneorisooctanemaybeusedtopreparesolutionsof2cyclohexylcyclohexanone(0,5 g/ml)(internal standard),2dodecylcyclobutanoneand2tetradecylcyclobutanone(10g/ml).Storeat20C. 4.7Florisil 3) , 150mto250m(60meshto100mesh),pesticideresidueanalysisgrade. Beforeuse,activatethe

    28、adsorbentbyheatingat550Cforatleast5horovernight.Coolinadesiccator.Keepwell sealedaftercooling. PreparedeactivatedFlorisil byadding20partsofwaterto100partsofadsorbent(m/m).Approximately30gof activatedFlorisil isrequiredtopreparesufficientdeactivatedadsorbentforeachcolumn.Ensurethatthe deactivatedFlor

    29、isil containsnolumpsandthatthepowderflowsfreely.Leavetoequilibrateovernight.Usewithin oneweek. 1) nHexanewasthesolventusedtovalidatethemethod.However,itisalsopossibletousenpentaneonhealth groundsprovideditcanbeshowntoleadtothesameresult. 2) ContacttheNationalStandardizationOrganizationsfortheavailab

    30、ilityofreferencestandards 3) Florisil isanexampleforasuitableproductavailablecommercially.Thisinformationisgivenfortheconvenienceof usersofthisstandardanddoesnotconstituteanendorsementbyCENofthisproduct.EN1785:2003(E) 5 4.8Nitrogen, forconcentratingsolutions 4.9Helium, ascarriergas 5Apparatus 5.1Gen

    31、eral Usuallaboratoryapparatusand,inparticular,thefollowing: 5.2Electricblender 5.3Soxhletapparatus, withsuitableflaskofe.g.250mlandextractorofe.g.100ml. 5.4Celluloseextractionthimbles, e.g.oflength80mmto100mm,withaninternaldiameterof30mm. Extractionwithnhexanepriortousemaybenecessary. 5.5Cottonwool,

    32、 nonabsorbent,washedinnhexanepriortouse. 5.6Electricheatingmantle or waterbath 5.7Chromatographictube, madeofglass,havingalengthof300mmandwithaninternaldiameterof20mm, fittedwithafrit,apolytetrafluoroethylene(PTFE)stopcockandagroundglassjointatthetop. 5.8Separatingfunnel, or droppingfunnel , e.g.of2

    33、50ml,withagroundglassjoint. 5.9Rotaryevaporator, withevaporationflaskandawaterbathcapableofbeingcontrolledat45C(low vacuum,approximately25kPa). 5.10 Apparatusforconcentrationofsolutions undernitrogen. 5.11 Gaschromatograph(GC)glassvials 5.12Gaschromatograph(GC) linkedtoa massspectrometer(MS). 5.13Ca

    34、pillarycolumn, withsuitableperformancecharacteristics,seeAnnexA.EN1785:2003(E) 6 5.14Laboratoryoven,capableofbeingmaintainedat100C 6 Samplingtechnique Whentakingsamples,givepreferencetothosepartsofthefoodwhichhaveahighfatcontent(e.g.chickenskin). Keepthesampleinasealableglassvesselorinafatfreemetalf

    35、oil. 7Procedure 7.1Samplepreparation Coarselychopthesamplesoffoodandthen,withtheexceptionofCamembert,homogenizeinanelectricblender (5.2).Camembertshouldbecutintosmallcubespriortoextraction.Forliquidwholeegg,ensurethatthesampleis thoroughlymixedpriortosampling. 7.2Fatextraction Weigh20gofanhydroussod

    36、iumsulfate(4.3)and20gofwellmixedhomogenizedsampleintoanextraction thimble(5.4),mixandplugwithcottonwool(5.5).Extrasodiumsulfatemaybeusedifnecessary.Forfoodstuffs havingalowfatcontent,itmaybenecessarytoincreasethesizeofthetestsampleandquantityofanhydrous sodiumsulfateaccordingly.Itisrecommendedthatli

    37、quideggisdriedat100Cfor12hpriortoextraction.Athin filmofeggpartiallydried(2hat100C)hasgivencomparableresults.Alternativedryingprocedures,e.g.freeze drying,maybeusedprovidedrecoveryof2alkylcyclobutanonesischecked(see7.6). Pour100mlofnhexane(4.2)intoasuitableflask(5.3)andplaceextractorontop.Placeextra

    38、ctionthimbleinthe extractorandadd40mlofnhexane.Placetheflaskontheheatingmantle(5.6)andcondenserontopofthe extractor.Refluxandextractgentlyfor6h.Thesolventshouldsiphonoverfourtimesinapproximately1h. Removetheflaskfromtheheatanddisposeofthethimbleandthenhexaneintheextractor.Transferthelipid extractfro

    39、mtheflasktoa100mlstopperedglassmeasuringcylinderandadjustthevolumeto100mlwithmore solvent.Add5gto10gofanhydroussodiumsulfate,stopper,mixandleaveovernight. Alternativefatextractionproceduresmaybeusediftheycanbeshowntoleadtothesameresults. 7.3Preparationofthelipidextract 7.3.1 Determinationoflipidcont

    40、entmethodI Dryduplicateflasksforatleast4horovernightat100C.Coolandweigh.Pipetteanaliquotoflipidextract(7.2) intoeachflaskandrotaryevaporate(5.9)todryness.Dryforatleast4horovernightat100Candreweigh. Alternatively,toprovideamorerapidmeasurementoflipidcontent,pipetteanaliquotoflipidextract(7.2)intoa pr

    41、eweighedglassvial.Evaporatethesolventunderastreamofnitrogen(5.10)toconstantweight.Calculatethe volumeofextractrequiredtoprovideapproximately200mgoflipid. 7.3.2 DeterminationoflipidconcentmethodII Alternativelyto7.3.1,concentratethewholelipidextract(7.2)toafewmillilitres(2mlto3ml)usingarotary evapora

    42、tor(5.9).Transfertheconcentratedlipidextracttoasmall(e.g.10ml)sealablepreweighedglassvial.Dry thesampleunderastreamofnitrogen(5.10)toconstantweight. 7.4Florisil columnchromatography PrepareaFlorisil column(20cmto21cm)usingachromatographictube(5.7),deactivatedFlorisil (4.7)andn hexane(4.2).Allowthenh

    43、exaneleveltodroptojustabovethetopoftheFlorisil . Takeavolumeoftheextract(7.3.1)whichprovidesapproximately200mgoflipidandconcentrateifnecessary. Thefinalvolumeshouldnotexceed5ml.Alternatively,dissolve200mgofthelipidobtainedin7.3.2innotmore than5mlofnhexane. Applythelipidextractquantitativelytothecolumn(recordexactweightoflipidapplied)andallowthenhexane l


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