SN 1544-2005進出口動物源性食品中玉米赤霉醇殘留量的檢驗方法 高效液相色譜-質譜質譜法

中華人民共和國出入境檢驗檢疫行業(yè)標準進出口動物源性食品中玉米赤霉醇殘留量的檢驗方法高效液相色譜-質譜/質譜法2005-02-17發(fā)布國家質量監(jiān)督檢驗檢疫總局I本標準的附錄A為資料性附錄。本標準由國家認證認可監(jiān)督管理委員會提出并歸口。本標準系首次發(fā)布的出入境檢驗檢疫行業(yè)標準。1進出口動物源性食品中玉米赤霉醇殘留量本標準規(guī)定了動物源性食品中玉米赤霉醇殘留量檢驗的抽樣、制樣和高效液相色譜-質譜/質譜檢以不超過2500件為一檢驗批。抽樣數(shù)量見表1。表1單位為件最低抽樣數(shù)126～1005101～250251～500501～10001001～25002.3抽樣方法按2.2規(guī)定的抽樣件數(shù)隨機抽取，逐件開啟。從每件中至少取一袋作為原始樣品。如每件中無小包裝或有小包裝，但每袋小包裝質量超過2kg,則可用滅菌刀具從每件中割取不少于500g,混合后置于清潔容器內(nèi)作為混合原始樣品?；旌显紭悠返目傊匈|量不少于2kg,加封后，標明標記，及時送交從混合原始樣品中取出部分有代表性的樣品，充分混勻，用四分法縮分至不少于500g,均分成兩3.1方法提要勻漿組織樣品經(jīng)葡萄糖苷酸酶水解后，依次用乙醚和氫氧化鈉溶液提取玉米赤霉醇。氫氧化鈉提取液中和后經(jīng)C??固相萃取柱凈化。甲醇洗脫液經(jīng)蒸發(fā)，定容后供高效液相色譜-質譜/質譜儀測定，外標法定量。23.2.7磷酸：純度>85%;密度1.7g/mL。3.2.8葡萄糖苷酸酶。酸調節(jié)至pH4.8。制成0.10mg/mL的標準儲備液。儲備液保存于-18℃。稱取5g試樣(精確至0.01g),置于潔凈的50mL具塞離心管中，加入10mL乙酸鈉緩沖溶液(3.2.13)和25μL葡萄糖苷酸酶，旋渦混勻，于37℃中保溫8h～14h。34玉米赤霉醇添加濃度及其回收率數(shù)據(jù)：——1pg/kg時，回收率72%;——5μg/kg時，回收率74%;5SN/T1544—2005(資料性附錄)0ThisstandardwasproposedbyandisunderthechargeofNationRegulatoryCommissionforCertifi-ThisstandardwasdraftedbyShanghaiEntry-ExitinspectionandQuarantineBureauofthePeople'sRepublicofChina.ThisstandardwasmainlydraftedbyFangXiaoming,ChenJiahua,NiXinluandTangYifeng.Thisstandardispromulgatedforthefirsttime.7productsforimportandexport—HPLC-MS/MSThisstandardspecifianimaloriginalproductsbyhigh-performanceliquidchromatography-tandemmassandspecification,shouldbTable1Minimumnumberofpackagestobetaken126～1005101～250251~500501～10001001～2500samplewasdevidedintwoandhomogenized.Packegeandlabel.8Thetestsampleshouldbestoredat-18℃.AhomogenizedtissuesampleishydrolyzedwithB-glucuronidaseandthehydrolysateisextractedwithethylether.Thesupernatantisevaporatedtodrynessandtheresidueisdissolvedinchloroformandre-extractedwithsodiumhydroxidesoCasolid-phaseextractioncartridge,andanalyzedbyelectrosprayHPLC-MS/MSinnegativeionmode.Reagentswereofanalytical-reagentgradeexceptforthespecificreagents.Thewaterusedwasdoublydistilledordeionizedw3.2.1Acetonitrile:LCgrade.3.2.6Anhydroussodiumacetate.3.2.7Phosphoricacid:purity>85%;3.2.9Zeranol(a-zearalanol):purity97%.withwater.3.2.13Sodiumacetatebuffer,50mmol/L:Take4.1gofanhydroussodiumacetate,dissolveand9diluteto1000mLwithwater.AdjustpHto4.8withglacialaceticacid.3.2.14Zeranolstocksolution,0.10mg/mL:Accuratelyweighproperzeranosolvedwithacetonitrile.Tstocksolution(3.2.14)into100mLvolumetrictively.ThesolutionsanolIntermediatesolutions(3.2.15).Thematrix-matchedstandardsarestoredat-week.3.2.17Cisolid-phaseextractioncartridge,500m3.3.1High-performanceliquidchromatography-electrospraytandemmassspectrometry.3.3.3VacuummanifoldforsolidWeigh5.0g±0.1gofhomogenizedtissuesampleintoa50mLcentrifugetube.Add10mLofsodi-SN/T1544—2005umacetatebuffer(3.2.13)and25μLofβ-glucuronidase,mixwell,andthenincubat(3.2.13)and2mLofmethan200pLofacetonitrileforanalb)Mobilephase:acetonitrile-water(40+60)containing0.025%aceticacid.d)Columntemperature:22℃.a)lonizationmode:negativeelectrospray.b)Conevoltage:50V.c)Capillaryvoltage:3kV.d)Collisionvoltage:20V.e)Analyzervacuum:<3mPa(3×10~5mbar).f)Collisiongas:argon.h)Nebulisergasflow:480L/h.i)Multiplereactionmonitoring(MRM)mode:m/z321→303,m/z321→277.m/forAnalyzematrix-matchedstandards(3.2.16)inorderofincreasingconcentration,obtainingthema-withoutadditionofthesample.Theconcentrationofzeranolinsampleiscalculatedwithchromatographicdataprocessoro…………(1)Where:X—theconcentrationofanalyteofinterestinsample,ug/kg:c~—theconcentrationofanalytefromcalibrationcurve,ng/mL;V--—finalvolumeofsampleextract,mL;m——weightofsample,g.Note:Theblankvalueshouldbesubtracte