SN1920-2007進(jìn)出口動(dòng)物源性食品中敵百蟲、敵敵畏、蠅毒磷殘留量的檢測(cè)方法液相色譜-質(zhì)譜質(zhì)譜法

中華人民共和國(guó)出入境檢驗(yàn)檢疫行業(yè)標(biāo)準(zhǔn)進(jìn)出口動(dòng)物源性食品中敵百蟲、敵敵畏、蠅毒磷殘留量的檢測(cè)方法液相色譜-質(zhì)譜/質(zhì)譜法Determinationoftrichlorfor,dichlorvosandcoumaphosresiduesinfoodstuffsofanimaloriginforimportandexport—LC-MS/MSmethod2007-05-23發(fā)布I本標(biāo)準(zhǔn)的附錄A、附錄B和附錄C均為資料性附錄。本標(biāo)準(zhǔn)由國(guó)家認(rèn)證認(rèn)可監(jiān)督管理委員會(huì)提出并歸口。本標(biāo)準(zhǔn)由中華人民共和國(guó)重慶出入境檢驗(yàn)檢疫局、中華人民共和國(guó)湖南出入境檢驗(yàn)檢疫局和中華人民共和國(guó)浙江出入境檢驗(yàn)檢疫局負(fù)責(zé)起草。本標(biāo)準(zhǔn)系首次發(fā)布的出入境檢驗(yàn)檢疫行業(yè)標(biāo)準(zhǔn)。1進(jìn)出口動(dòng)物源性食品中敵百蟲、敵敵畏、蠅毒磷殘留量的檢測(cè)方法液相色譜-質(zhì)譜/質(zhì)譜法質(zhì)譜/質(zhì)譜檢驗(yàn)方法。2試樣的制備與保存分割肉：取樣品中有代表性的約500g,用組織搗碎機(jī)搗碎，裝入潔凈容器作為試樣，密封并做好標(biāo)腸衣：取有代表性樣品約100g,用剪刀剪碎至2mm以下，裝入潔凈容器作為試樣，密封并做好標(biāo)識(shí)，于-18℃冰箱內(nèi)保存。蜂蜜：取有代表性樣品約500g,攪拌均勻后裝入潔凈容器內(nèi)密封并做好標(biāo)識(shí)，于一18℃冰箱內(nèi)保存。制樣操作過(guò)程中應(yīng)防止樣品受到污染或發(fā)生殘留物含量的變化。3方法提要4試劑和材料4.3乙酸乙酯。4.4二氯甲烷。4.5環(huán)己烷。4.6硫酸鈉：650℃灼燒4h,在干燥器內(nèi)冷卻至室溫，貯于密封瓶中備用。4.850mmol/L乙酸銨溶液：稱取0.385g乙酸銨溶于1000mL水中。4.9敵百蟲(Trichlorfor,CAS號(hào)：52-68-6,分子式：C?H?Cl?O?P)標(biāo)準(zhǔn)品：純度大于或等于98.0%。4.10敵敵畏(Dichlorvos,CAS號(hào)：62-73-7,分子式：C?H?Cl?O?P)標(biāo)準(zhǔn)品：純度大于或等于97.6%。4.11蠅毒磷(Coumaphos,CAS濃度的混合標(biāo)準(zhǔn)儲(chǔ)備液。此溶液在0℃~4℃冰箱中避光保存，可使用3個(gè)月。4.13空白樣品提取液：用不含敵百4.14標(biāo)準(zhǔn)工作溶液：根據(jù)需要用空白樣品提取液將標(biāo)準(zhǔn)儲(chǔ)備液稀釋成50ng/mL、100ng/mL、2200ng/mL、500ng/mL的混合標(biāo)準(zhǔn)工作溶液。置于0℃~4℃冰箱中避光保存，可使用3d。5儀器和設(shè)備5.1高效液相色譜-質(zhì)譜/質(zhì)譜儀：三重四極桿質(zhì)譜檢測(cè)器，配電噴霧離子源(ESI)或相當(dāng)者。5.2超聲波清洗器。5.3旋渦混勻器。5.4旋轉(zhuǎn)蒸發(fā)儀。5.6均質(zhì)器。5.70.45μm微孔濾膜。6.1提取與凈化6.1.1分割肉a)準(zhǔn)確稱取5g均勻試樣(精確到0.01g)于50mL具塞試管中，加入5g無(wú)水硫酸鈉混勻，再加入15mL二氯甲烷，用均質(zhì)器(10000r/min)均質(zhì)2min,4000r/min離心3min,將有機(jī)相轉(zhuǎn)移至100mL梨形蒸餾瓶中，殘?jiān)儆?×10mL二氯甲烷均質(zhì)提取兩次。b)離心合并有機(jī)相，于40℃旋轉(zhuǎn)蒸發(fā)至2mL,將樣液轉(zhuǎn)移至5mL刻度試管中，并用少量二氯甲烷洗滌梨形蒸餾瓶，合并洗滌液到刻度試管中，室溫通氮濃縮至干。定量加入1.0mL乙腈溶解殘?jiān)?，加?mL環(huán)己烷旋渦混勻2min后，2500r/min離心3min,將下層乙腈相過(guò)6.1.2腸衣準(zhǔn)確稱取5g試樣(精確到0.01g)于50mL具塞離心瓶中，加入2g無(wú)水硫酸鈉混勻，再加入15mL二氯甲烷，蓋上蓋混勻，置于超聲波清洗器中超聲30min,冷卻后將有機(jī)相過(guò)濾轉(zhuǎn)移至100mL梨形蒸餾瓶中，殘?jiān)儆?×10mL用二氯甲烷混勻超聲提取兩次，離心合并有機(jī)相。接下來(lái)按照6.1.1中b)的步驟進(jìn)行。稱取5g試樣(精確到0.01g)置于50mL具塞離心管中，加10mL水，加25mL乙酸乙酯，于旋渦混合器上混勻1min,以3000r/min離心5min,將上層乙酸乙酯提取液收集于濃縮瓶中，殘?jiān)偌尤?0mL乙酸乙酯，重復(fù)上述操作，合并乙酸乙酯提取溶液。在50℃以下減壓濃縮至約3mL后轉(zhuǎn)移至離心管，用5mL乙酸乙酯分兩次洗滌濃縮瓶，合并洗滌液于離心管中，用氮?dú)獯蹈筛伞＜?.0mL乙腈溶6.2.1液相色譜-質(zhì)譜/質(zhì)譜條件a)色譜柱：柱填料為十八烷基硅烷鍵合相的色譜柱，4.6×150mm,5μm或相當(dāng)者；i)質(zhì)譜/質(zhì)譜參考條件參見附錄B。36.2.2定性、定量測(cè)定按照6.2.1液相色譜-質(zhì)譜/質(zhì)譜條件測(cè)定樣液和標(biāo)準(zhǔn)工作溶液，外標(biāo)標(biāo)準(zhǔn)曲線法測(cè)定樣液中的敵百蟲、敵敵畏、蠅毒磷殘留量。樣品中待測(cè)物殘留量應(yīng)在標(biāo)準(zhǔn)曲線范圍之內(nèi)，如果殘留量超出標(biāo)準(zhǔn)曲線范圍，應(yīng)用空白樣品提取液(4.13)進(jìn)行適當(dāng)稀釋。在上述色譜條件下，敵百蟲、敵敵畏、蠅毒磷的質(zhì)量色譜峰保留時(shí)間約為13.2min,14.1min,15.9min。標(biāo)準(zhǔn)品的多反應(yīng)監(jiān)測(cè)色譜圖參見附錄C中的在相同的實(shí)驗(yàn)條件下，樣品與標(biāo)準(zhǔn)工作液中待測(cè)物質(zhì)的質(zhì)量色譜峰相對(duì)保留時(shí)間在2.5%以內(nèi)，并且在扣除背景后的樣品質(zhì)量色譜圖中，所選擇的離子對(duì)均出現(xiàn)，同時(shí)與標(biāo)準(zhǔn)品的相對(duì)豐度允許偏差不超過(guò)表1規(guī)定的范圍，則可判斷樣品中存在對(duì)應(yīng)的被測(cè)物。表1使用定性液相色譜-質(zhì)譜/質(zhì)譜時(shí)相對(duì)離子豐度最大容許誤差相對(duì)豐度(基峰)/(%)液相色譜-質(zhì)譜/質(zhì)譜時(shí)相對(duì)離子豐度最大允許誤差/(%)大于20至小于等于50大于10至小于等于20小于等于107結(jié)果計(jì)算和表述用數(shù)據(jù)處理軟件中的外標(biāo)法，或繪制標(biāo)準(zhǔn)曲線，按照式(1)計(jì)算樣品中敵百蟲、敵敵畏、蠅毒磷殘留量。……式中：X——試樣中敵百蟲、敵敵畏、蠅毒磷的殘留量，單位為毫克每千克(mg/kg);c——由標(biāo)準(zhǔn)曲線而得的樣液中敵百蟲、敵敵畏、蠅毒磷的濃度，單位為納克每毫升(ng/mL);V——樣液最終定容體積，單位為毫升(mL);m——最終樣液所代表的試樣質(zhì)量，單位為克(g)。8測(cè)定低限、回收率8.1測(cè)定低限本方法敵百蟲、敵敵畏、蠅毒磷的測(cè)定低限均為0.010mg/kg。8.2回收率8.2.1腸衣中敵百蟲添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在73.0%～94.0%之間；添加濃度在0.020mg/kg時(shí)，回收率在78.0%～95.0%之間；添加濃度在0.040mg/kg時(shí)，回收率在78.0%～95.0%之間。8.2.2腸衣中敵敵畏添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在76.0%～98.0%之間；添加濃度在0.020mg/kg時(shí)，回收率在78.0%～98.0%之間；添加濃度在0.040mg/kg時(shí)，回收率在86.0%～95.0%之間。8.2.3腸衣中蠅毒磷添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在75.0%～95.0%之間；添加濃度在0.020mg/kg時(shí)，回收率在80.0%～96.0%之間；4添加濃度在0.040mg/kg時(shí)，回收率在77.0%～92.0%之間。8.2.4分割肉中敵百蟲添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在92.0%～115%之間；添加濃度在0.020mg/kg時(shí)，回收率在86.5%～113%之間；添加濃度在0.040mg/kg時(shí)，回收率在80.8%～101%之間。8.2.5分割肉中敵敵畏添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在93.0%～110%之間；添加濃度在0.020mg/kg時(shí)，回收率在88.5%～108%之間；添加濃度在0.040mg/kg時(shí)，回收率在84.3%～110%之間。8.2.6分割肉中蠅毒磷添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在91.0%～112%之間；添加濃度在0.020mg/kg時(shí)，回收率在79.0%～102%之間；添加濃度在0.040mg/kg時(shí)，回收率在73.9%～95.3%之間。8.2.7蜂蜜中敵百蟲添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在70.0%～110%之間；添加濃度在0.020mg/kg時(shí)，回收率在79.0%～104%之間；添加濃度在0.040mg/kg時(shí)，回收率在80.0%～107%之間。8.2.8蜂蜜中敵敵畏添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010mg/kg時(shí)，回收率在70.0%～92.0%之間；添加濃度在0.020mg/kg時(shí)，回收率在71.0%～91.0%之間；添加濃度在0.040mg/kg時(shí)，回收率在70.0%～96.0%之間。8.2.9蜂蜜中蠅毒磷添加濃度及其回收率數(shù)據(jù)：添加濃度在0.010添加濃度在0.020添加濃度在0.040mg/kgmg/kgmg/kg時(shí)，回收率在79.0%～110%之間；時(shí)，回收率在74.0%～105%之間；時(shí)，回收率在71.0%～104%之間。5(資料性附錄)梯度洗脫程序表A.1流動(dòng)相梯度洗脫程序時(shí)間/min甲醇/(%)50mmol/L乙酸銨/(%)090.090.095.095.090.025.0090.0(資料性附錄)API4000LC-MS/MS系統(tǒng)電噴霧離子源參考條件：a)窗簾氣(CUR):15.00Pa;b)霧化氣(GS1):40.00Pa;c)輔助加熱氣(GS2):45.00L/min;d)碰撞氣(CAD):7.00Pa;e)離子源噴霧器電壓(IS):5000.00V;g)定性離子對(duì)、定量離子對(duì)、去簇電壓、碰撞能量、碰撞室出口電壓見表B.1。表B.1待測(cè)物定性離子對(duì)、定量離子對(duì)、去簇電壓、碰撞能量和碰撞室出口電壓待測(cè)物m/zm/z去簇電壓/V碰撞能量/V碰撞室出口電壓/V敵百蟲Trichlorfon259.0°63.026.5256.9221.263.0敵敵畏Dichlorvos221.3°63.025.5223.363.025.0蠅毒磷Coumaphos363.3“307.096.025.020.0363.3227.096.0a為定量離子對(duì)。6(資料性附錄)標(biāo)準(zhǔn)品多反應(yīng)監(jiān)測(cè)色譜圖3000001ARM|30=0.252011022aruhonSmon00B1-u)150A??11.四疊加離子色譜圖Kax.63a*.敵百蟲259.0→109.220XEalIKRM|7D*0.221211039amuhonFanOamD0I-00100XCal1MRM130*0.262319032amu""iomFandelad1Hom0s1200811-2a*Iuo)2敵敵畏221.3→109.27Kax.4,Fm*0蠅毒磷363.3→307.02圖C.1敵百蟲、敵敵畏、蠅毒磷標(biāo)準(zhǔn)溶液多反應(yīng)監(jiān)測(cè)色譜圖7AnnexA,BandCofthisstandardareinformativeannexes.tificationandAccreditationofthePeoplesRepublicofChina.ThestandardwasdraftedbyChongqingEntry-ExitInspectionandQuarantineBureauofthePeople'sna,andZhejiangEntry-ExitInspectionandQuarantineBureauofthePeople'sRepublicofChina.ThisstandardwasmainlydraftedbyWangGuoming,DaiHua,XieWen,ZhuGe,WangMeiling,LiXianliang,LiYingguo,ZhangLei,ZhouQimingandWangXiangxian.Thisstandardisaprofessionalstandardforentry-exitinspectionandquarantinepromulgatedforthefirsttime.Note:ThisEnglishversion,atranslationfromtheChinesetext,isonlyforreference.8byliquidchromatography-massspectrometryoftrichloffor,dichlorvosandcoumaphosresiduesinanimalmuscle,saltedcasingandhoney.Thisstandardisapplicabletothedeterminationoftrichloffor,dichlorvosandcoumaphosresiduesinanimalmuscle,saltedcasingandhoney.Muscle:About500grepresentativesamplesshouldbetakenfromallsamples,thengrindedandblen-dedbyatissueblendertoproducehomogenoussamples,putinsuitablecleancontainer.Afterbeingsealedandlabeled,thesamplesshouldbestoredatbelow-18Casing:About100grepresentativesamplesshouldbetakenfromallsamples,thengrindedandblen-dedbyaforfex,putinsuitablecleancontainer.Afterbeingsealedandlabeled,thesamplesshouldbeHoney:About500grepresentativesamplesshouldbetakenfromallsamples,thenmixroundtopducehomogenoussamples,putinsuitablecleancontainer.Afterbeingsealedandlabeled,thesamplesshouldbestoredatbelow-18℃.Inthecourseofsamplingandsamplepreparation,precautionshouldbetakentoavoidcontaminationoranyfactorthatmaycausethechangeofresiduecontent.EthylAcetateordichloromethane.Afterbeingconcentratedandreconstituted,theresiduesaredeter-minedbyliquidchromatography-massspectrometry,quantifiedbyexternalstandardmethod.Unlessspecified,allreagentsshouldbeofanalyticalgrade;“Water”isthedistilledwater.94.2Methanol:HPLCgrade.4.4Dichloromethane.4.5Cyclohexane.4.6Sodiumsulfate:lgniteat650℃for4h,cooltoroomtemperatureinadesiccatorandstoreinsealedcontainer.4.7Ammoniumacetate:GR.4.850mmol/Lammoniumacetate:Weigh0.385gammoniumacetate,dissolvedin1000mLwa-ter.4.9Trichlorfor(CASNO:52-68-6formula:C?H?Cl?4.11Coumaphos(CASNO:56-72-4,formula:C?Hi?CIO?PS),purity≥99.9%.4.12Standardstocksolution:Accuratelyweightrichlorfor,dichlorvoswithmethanol,andmixtohomogeneity.Theconcentrationofthesolutionis100g/mL,canbepre-servedatthetemperature0℃~4℃forthreemonths.chlorvosandcoumaphos.4.14Standardworkingsolution:accordingtotherequirement,dilutemixedstandardsolutionwithblankmatrixextractsolution.Theconcentrationofthesolutionis50ng/mL,100ng/mL,200ng/mLand500ng/mL.Itcanbepreservedatthetemperature0℃~4℃forthreedays.5Apparantusandequipme5.1Liquidchromatography-massspectrometry,equippedwithelectrosprayionsourceandtriqua-5.2UItrasonicextractor.5.3Mechanicalshaker.SN/T1920—20075.5Centrifuge:5000r/min.5.6Homogenizer.5.7Membranefilter:0.45μm.a)Accuratelyweigh5gofthetestsample(accurateto0.01g)intoa50mLpolypropylenecentri-fugetubewithcap,add5ganhydroussodiumsulfateand15mLofdichloromethane,homogenizefor2min(10000r/min),thencentrifugeat4000r/minfor3min.Transferthesupematanttoa100mLevaporatedflask,theresiduewashomogenizedandextractedwith2×10mLdichlo-romethane.b)Aftercentrifuging,combinethesupernatanttotheevaporatedflask.Evaporatethedichlo-romethanetoabout2mLwithrotaryevaporatorbelow40℃,removetheresiduestoa5mLcentrifugetube,washevaporatedflaskwithlittledichloromethane,combinethedichloromethanesolutiontograduatedtesttube,evaporatetheextracttodrynessonN-Evapatroomtempera-ture.Theresidueisdissolvedwith1.0mLofacetonitrileand2mLofcyclohexane.Votexandcentrifugeat2500r/minfor3min.Filterthenetherlayerthrougha0.45μmmembraneintoHPLCvials.ThesolutionisreadyforLC-MS/MSdetermination.Accuratelyweigh5gofthetestsample(accurateto0.01g)intoa50mLpolypropylenecentrifugetubewithcap,add5ganhydroussodiumsulfate,add15mLofdichloromethaneandvortex,thenex-tractinultrasonicextractorfor30min.Aftercooling,transferthesupematanttoa100mLevaporat-edflask,theresiduewasvortexedandextractedwith2×10mLdichloromethane.Aftercentrifuging,combinethesupernatanttotheevaporatedflask.Then,dealwiththesampleaccordingtothestepof6.1.1b).Accuratelyweigh5gofthetestsample(accurateto0.01g)intoa50mLpolypropylenecentrifugetubewithcap,add10mLwaterand25mLethylacetate,shakefor1minonamechanicalshakerat2000r/min,thencentrifugeat3000r/minfor5min.Transferthesupernatanttoaevaporatedflask,theresidueswasre-extractedwith20mLethylacetateastheaboveprocedure,combinethesuper-natant,evaporatedtoabout3mLwithrotaryevaporatorinawater-bathbelow50℃,removetheres-iduetoanewcentrifugetube,washtheevaporatedflasktwiceuse5mLethylacetate,combinetheeluate,evaporatetheeluatetodrynessonN-Evap,dissolvedby1.0mLofmethanol,FilterthroughaSN/T1920—20070.45μmmembraneintoHPLCvials.ThesolutionisreadyforLC-MS/MSdetermination.6.2.1LC-MS/MSoperationconditiona)LCcolumn:Ca,4.6×150mm,5μm(orotherconformablecolumn);b)Mobilephase:methanol:50mmol/Lammoniumacetate,theelutiongradientislistedinAnnexA;d)Columntemperature:40℃;e)Injectorvolume:10μL;f)lonsource:ESl;g)Scanningmodel:positiveion;h)Monitoringmodel:multiplereactionmonitoring(MRM);i)ReferencedconditionsseenAnnexB.6.2.2QualificationandquantificationtestAccordingtothemethodof6.2.1,detecttheresiduesoftrichlorfor,dichlorvosandcoumaphosinthetestsamplesolution,thestandardworkingsolution.Theresponseoftrichlorfor,dichlorvosandcoumaphosshouldbeinthelinearrangeoftheinstrumentaldetection.Iftheresponseisoutofthelinearrange,thesampleshouldbedilutedwiththeblankmatrixextractsolution(4.13)tosuitableconcertration.Undertheabovechromatographconditions,thereferenceretentiontimeoftrichlorfor,dichlorvosandcoumaphosisabout13.2min,14.1min,15.9min.ReconstitutedionchromatogramofstandardworkingsolutionislistedinAnnexC.Underthesameconditionsofexperiment,theretentiontimeoftheunknownsampleisthesameasthestandardworkingsolution;thequalificationionsforeverycompoundmustbefound.Forthesameanalysisbatchandthesamecompound,thevariationrangeoftheionratiobetweenthetwodaughterionsfortheunknownsampleandthestandardworkingsolutionatthesimilarconcentrationcannotbeoutofrangeoftable1.Table1—MaximumpermittedtolerancesforrelativeionintensitieswhileconfirmationRelativeintensity/(%)Permittedtolerances/(%)7CalculationandexpressionofresultCalculatingthecontentoftrichlorfor,dichlorvosandcoumaphosresidueconcentrationinthesampleiscarriedoutbyLC/MS/MSdataprocessororaccordingtotheformula(1): (1)c—theconcentrationoftrichlorfor,dichlorvosandcoumaphosinthetestsamplecalculatedbycali-brationcurve,ng/mL;m—thecorrespondingmassoftestsampleinthefinalsamplesolution,g;V—thefinalvolumeofsamplesolution,mL.8.1LimitofquantificationThelimitofquantificationfortrichlorfor,dichlorvosandcoumaphosis0.010mg/kg8.2.1Accordingtotheexperimentaldate,thefortifyingconcentrationoftrichlorforincasinganditscorrespondingrecoveriesare:0.010mg/kg,therecoveryis73.0%～94.0%;0.020mg/kg,therecoveryis78.0%～95.0%;0.040mg/kg,therecoveryis78.0%～95.0%.8.2.2Accordingtotheexperimentaldate,thefortifyingconcentrationofdichlorvosincasinganditscorrespondingrecoveriesare:0.010mg/kg,therecoveryis76.0%～98.0%;0.020mg/kg,therecoveryis78.0%～98.0%;0.040mg/kg,therecoveryis86.0%～95.0%.8.2.3Accordingtotheexperimentaldate,thefortifyingconcentrationofcoumaphosincasinganditscorrespondingrecoveriesare:0.010mg/kg,therecoveryis75.0%～95.0%;0.020mg/kg,therecoveryis80.0%～96.0%;0.040mg/kg,therecoveryis77.0%～92.0%.8.2.4Accordingtotheexperimentaldate,thefortifyingconcentrationoftrichlorforinmusleanditscorrespondingrecoveriesare:0.010mg/kg,therecoveryis92.0%～115%;0.020mg/kg,therecoveryis86.5%～113%;0.040mg/kg,therecoveryis80.8%～101%.SN/T1920—20078.2.5Accordingtotheexperimentaldate,thefortifyingconcentrationofdichlorvosinitscorrespondingrecoverie0.010mg/kg,therecoveryis93.0%～110%;0.020mg/kg,therecoveryis80.040mg/kg,therecoveryis84.3%～110%.8.2.6Accordingtotheexperimentaldate,thefortifyingconcentrationofcoumaphosinitscorrespondingrecoverie0.010mg/kg,therecoveryis91.0%～112%;0.020mg/kg,therecoveryis79.0%～102%;0.040mg/kg,therecoveryis73.9%～95.3%.8.2.7Accordingtotheexperimentaldate,thefortifyingconcentrationoftrichlorforinitscorrespondingrecoverie0.010mg/kg,therecoveryis70.0%～110%;0.020mg/kg,therecoveryis79.0%～104%;0.040mg/kg,therecoveryis80.0%～107%.8.2.8Accordingtotheexperimentaldate,thefortifyingconcentrationofdichlorvosinitscorrespondingrecoverie0.010mg/kg,therecoveryis70.020mg/kg,therecoveryis70.040mg/kg,therecover