SNT 2451-2010动物源性食品中呋喃苯烯酸钠残留量检测方法液相色谱-质谱质谱法

中华人民共和国出入境检验检疫行业标准SN/T2451—2010动物源性食品中呋喃苯烯酸钠残留量检测方法液相色谱-质谱/质谱法2010-01-10发布2010-07-16实施I1SN/T2451—2010动物源性食品中呋喃苯烯酸钠残留量检测方法液相色谱-质谱/质谱法2规范性引用文件GB/T6682分析实验室用水规格和试验方法(GB/T6682—2008,ISO3696:1987,MOD)5.5呋喃苯烯酸钠标准品(Ci₃HgNNaO₅):CAS登录号为54992-23-3;纯度大于等于99.0%,购自Wakepurechemicalindustries5.70.1%甲酸溶液：取1.0mL甲酸用水定容至1000mL。5.8溶解液：0.1%甲酸溶液十乙腈(4+1,体积比)。25.10标准储备溶液：1.0mg/mL,取25mg标准品用少量甲醇溶解并定容至25mL,-18℃避光现用。下同7.1.1。向溶解液中加入3mL乙腈饱和正己烷(5分),稍加振摇后静置分层后，取下层溶液于9500r/min离心5min,供液相色谱-质谱/质谱检测。表1梯度洗脱程序时间/min流动相比例A%B%3SN/T2451—2010表1(续)时间/min流动相比例A%B%7.3.2质谱条件d)其他条件参见附录A。8.1标准曲线的绘制用空白基质溶液将呋喃苯烯酸钠标准中间工作溶液(5.11)逐级稀释得到2ng/mL、5ng/mL、酸钠。相对离子丰度/%>10～20允许的最大偏差/%±士304 本方法在5.0pg/kg、10.0pg/kg、20pg/kg添加水平的回收率及精密度参见附录C中表C.1.5质谱/质谱测定参考条件1)表A.1呋喃苯烯酸钠质谱/质谱多反应监测条件化合物母离子子离子破裂电压/V碰撞电压/V驻留时间/s呋喃苯烯酸钠258.3214.0⁴5a离子用于定量。67(资料性附录)LC-MS/MS法添加回收率及精密度表C.1LC-MS/MS法添加回收率及精密度样品名称添加浓度/(μg/kg)回收率/%相对标准偏差/%猪肉579.6～90.04.381.9～87.083.5～87.5猪肝582.4～87.081.9～86.878.0~93.0猪肾585.0～99.281.2～90.24.482.5～90.0鳗鱼580.0～86.281.5≈86.3鸡蛋576.886.482.0～84.2牛奶577.~86.45～86.58ficationandAccreditation.ThisstandardwasdraftedbyJiaRepublicofChina.ThismaindraftersofthisstandardareZhanChunrui,WenZhihai,GuoPing,LiHaiyan,ZhouPingyong,GuiJiaxiang.9Thisstandardisapplicabletothedeterminationofsodiumnifurstyrenateresiduesinliver,kidneypork,eel,eggandmilk.Thefollowingnormativedocumentscontainprovisionswhich,throughreferenceinthistext,consti-tuteprovisionsofthisprofessionalstandard.Fordatedreferences,subsequentamendmentsto,orre-visionsof,anyofthesepublicationsdonotapply.However,partiefessionalstandardareencouragedtoinvestigatethepossibilityofapplyingthemostrofthenormativedocumentsindicatedbelow.Forundatedreferencetivedocumentreferredtoapplies.GB/T6682Waterforanalyticallaboratoryuse—Specificationandtestmethods(GB/T6682—2008,IS03696:1987,MOD)3Samplepreparationandsto3.1Topork,liverandkidneysamples,about500greprestionofthetestsampleisputintoacleancontainer,thensealandlabelit.Thesamplesarestoredbe-low-18℃.Pleasecoo3.2Toeggs,pealofftheskin,mixthroughlyandthendivideintotwoequalportions.Eachportion-18℃.Pleasecoolatroomt3.3Tomilk,stirmixthroughlyandthendivideintotwoequalportions.EachportionofthetestPleasecoolatroomtemperaturewhenuse.Thesodiumnifurstyrenateresiduesinthetestsamplesareextractedwithacetonitrile,thenpurifedbyliquid-liquidpartitionwithsaturatedhexaneinacetonitrile,afterconcentratedanddiluted.Thefi-method.Unlessotherwisespecified,allregents.are.analyticallypure,“water?isdeionizedwater.5.1Methanol:LCgrade.5.2Acetonitrile:LCgrade.5.3Formicacid:LCgrade.5.4n-hexane:LCgrade.5.5Sodiumnifurstyrenatestandard(CgHaNNaO₅):CAS54992-23-3,purity≥99.0%.Buyingfrom5.6Anhydroussodiumsulfate:Igniteat656Cfor.2h,andkeepinasolvetovolumewithacetonitrile.5.8Combinationsolution:0.1%Formicacidinwater+acetonitrile=4+1(V/V).oughly,drawabovehexane.5.10Standardstocksolution:1.0mg/mL,Accuratelyweigh25mginto25mLvolumetricflack,dis-solvetovolumewithmethanol,andmixtohomogeneity.Thesolutioncanbestoredatthetempera-5.11Standardworkingsolution:10.0μg/mL,Accuratelyweigh(5.10)into10mLvolumetric,di-lutewithmethanoljustbeforeuse.6.4Centrifuger:4000r/min.9500r/min.6.5Colorimetrictube:50mL.6.7Nitrogenblowingdevice7.1.1Pork,liver,kidney,egg,eelWeigh5g(accurateto0.01g)ofthetestsampleintoa50/mLcentrifugetube,add25mLofaceto.nitrileandmixfor30s,thencentrifugefof5minat4000f/min.Theuppers50mLcolorimetrictube,repedttheaboveprocedureohce,combinedtheextractionsolution,dilutedevicewithabathtemperaturebelow.40C.DissolvWeigh5g(accurateto0.01g)ofthetestsampleintoa50mLcentrifugetube,andabout5ganhydroussodiumsulfate,mixfully.Theotherstepwassimilarto7.1.1.cardhexaneaftercentrifugefor5minat9500r/min,forLC-MS/Ma)Chromatographiccolumn:PhenomenexLunaCia,150mm×2.1mm(i.d.),3pm,orequivalent;e)Injectorvolume:20μL.Time/minRationofthemobileA/%B/%b)Scanmode:Negitive;c)Monitoringmodel:Multiplereactionmonitoring(MRM)d)TheotherconditionWeigh5g(accurateto0.01g)8LC-MS/MSdeterminationDilutestepwisethestandardworkingsolution(5.11)tomatrixstandardsolutionswithconcentra-tionsof2、5、10、20and50ng/mLbyblankmatrixsolution.AccordingtotheLC-MS/MSconditiona-bovementioned,determinatedthematrixstandardsolutionsfromlowtohigh.Drawthestandardcurve,abscissapresentstheconcentrationofmatrixstandardsolutionsandordinatepresentsre-figureB.1.taintimeofsodiumnifurstyrenateis8.3min.Underthesamedeterminationcondition,theratioofthechromatographicretentiontimeoftheanalystsshallcorrespondtothatofthecalibrationsolutionatatoleranceof±2.5%.Therelativeintensitiesofthedetectedionsofeachanalysts,shallcom-poundtothoseofthecalibrationstandardatcomparableconcentrations,withinthetolerancesshownintable2,thenthecorrespondingforrelativeionintensitieswhileconfirmation.Relativeintensity/%>20～50>10～20Permittedtolerances/%Accordingtotheapproximateworkingoperatingcondition,theresponseofsodiumnifurstyrenatethesampleshouldbedilutedwiththeblankmatrixsolutiontosuitableconcentratin.Calculatethecontentoffollowedformul