【行业标准】 农产品中甲萘威、毒死蜱、霜霉威、甲霜灵、甲草胺、异丙草胺残留胶体金快速检测方法

中华人民共和国出入境检验检疫行业标准SN/T 2921—2011
农产品中甲蔡威、毒死蜱、霜霉威、甲霜灵、甲草胺、异丙草胺残留胶体金快速检测方法
Fast determination of carbaryl,chlorpyrifos,propamocarbmetalaxyl,alachlor,metolachlor in farm produces bycolloidalgoldimmunoassay
2011-05-31发布
中华人民共和国
国家质量监督检验检疫总局
2011-12-01实施
本标准按照GB/T1.1一2009给出的规则起草本标准由国家认证认可监督管理委员会提出并归口SN/T2921—2011
本标准起草单位：中华人民共和国天津出入境检验检疫局、中华人民共和国湖南出入境检验检疫局、北京万华生物工程有限公司。本标准主要起草人：万积成、万志静、宓捷波、莫谨、郑文杰、唐晓宾、营丽萍。I
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1范围
农产品中甲蔡威、毒死蜱、霜霉威、甲霜灵、甲草胺、异丙草胺残留胶体金快速检测方法
SN/T 2921—2011
本标准规定了农产品中甲茶威（carbaryl）、毒死蜱（chlorpyrifos）、霜霉威（propamocarb）、甲霜灵（metalaxyl）甲草胺（alachlor）、异内草胺（metolachlor）的胶体金快速检测方法。本方法适用于黄瓜、番茄、西兰花、白菜、梨果等蔬菜和水果中甲蔡威、毒死蜱、霜霉威、甲霜灵、甲草胺、异丙草胺残留量的筛选检验,阳性结果须用其他方法进行确证2规范性引用文件
下列文件对于本文件的应用是必不可少的。凡是注日期的引用文件，仅注日期的版本适用于本文件。凡是不注日期的引用文件，其最新版本（包括所有的修改单）适用于本文件。GB/T6682分析实验室用水规格和试验方法3原理
以大孔径微孔滤膜为载体，胶体金为固相标记物，制作检测条。利用免疫测定中的双抗体夹心法或竞争法检测原理·通过在检测区包被一定量的单抗（双抗体夹心）或抗原（竞争法），对照区包被抗鼠多克隆抗体，利用胶体金标记的抗体与样品及检测区的物质之间的夹心结合或竞争反应，观测在检测条的检测区和对照区呈现的反应结果（检测条出现色带），然后根据色带出现的条数，判断农药检出或超标。
4试剂与材料
除另有规定外，所有试剂均为分析纯和GB/T6682中规定的三级水。4.1氯化钠，
4.2胶体金免疫检测试剂盒：分为甲萘威、毒死蜱、霜霉威、甲霜灵、甲草胺、异丙草胺的胶体金检测试剂盒，北京万华生物工程有限公司产品1，各类试剂盒中包括阳性参考品、阴性参考品和检测试剂条·检测试剂条使用铝箔袋密封包装。5仪器与设备
5.1均质器。
5.2培养血。
1）此处列出这一信息是为了方便本标准的使用者，生产厂家仅为提供参考，并不涉及商业目的，鼓励标准使用者尝试不同厂家或型号的试剂盒。
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6试样制备
称取5g（精确至0.1g）水果或蔬菜，擦去表面泥土，剪成1cm2左右碎片后，加入0.5g氯化钠，放人均质器中绞碎，静置3min～5min，取汁液置于培养Ⅲ内作为样品处理液待用。7测定
撕开铝箔袋，取出检测试剂条。将检测试剂条取样端浸入样液中（液面不得超过取样最高线）。样液在毛细管作用下向检测试剂条的另一端缓缓移行，在1min5min（最长不超过5min）之内观察检测区和对照区的色带条数和颜色深浅。同时对试剂盒内的阳性参考品、阴性参考品和水（空白）进行同样操作，以协助结果判断。8结果判定与表述
对照阴性参考品、阳性参考品和空白的结果，根据双抗体夹心和竞争法的区别及检测试剂条检测区和控制区的色带，按照如下原则进行结果判定：a）无效：在检测试剂条的控制区未出现色带：阳性：在检测区及控制区各出现一条色带（双抗体夹心法检测条）；b)
仅在控制区出现一条红线（竞争法检测条）：c
阴性：仅在控制区出现一条色带（双抗体夹心法检测条）；在检测区及控制区各出现一条红线（竞争法检测条）；d）临界值：在控制区出现一条红线，在检测区出现一条模糊的阴影线（竞争法检测条）。检测条检测结果判断图参见附录A。凡是结果呈现阳性或临界值的样品均为“阳性可疑”样品，需重复检测两次以上，并可用质谱方法进行确证。
9检测限和特异性
9.1检测限
表1检测限
农药名
甲萘威
毒死蜱
霜霉威
甲霜灵
甲草胺
异丙草胺
农产品类别
白菜等
番茄、甘蓝、西兰花等
黄瓜、葡萄等
白菜等
黄瓜、菜心等
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检测条检测限
2特异性
阳性样品的符合率≥90%。
阴性样品的符合率≥90%。
假阴性率≤5%。
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邻界值
附录A
（资料性附录）
检测条检测结果判断图2
双抗体夹心法检测条结果判断示意图反
-- MAX
控制线
竞争法检测条结果判断示意图
2）非商业性声明：附录A所列检测条检测结果判断图是采用北京万华生物工程有限公司胶体金试剂盒进行试验所得的结果示例，此处列出生产厂家仅为提供参考，并不涉及商业目的，鼓励标准使用者尝试不同厂家或型号的试剂盒。TTKANY KAa
Foreword
ThisstandardwasdraftedinaccordancewiththeGB/T1.1-2009SN/T2921—2011
This standard was proposed by and is under the jurisdiction of the Certification and AccreditationAdiministrationof thePeople'sRepublicof China.This standard was mainly drafted by Tianjin Entry-Exit Inspection and Quarantine Bureau of the Peo-ple's Republic of China.Hunan Entry-Exit Inspection and QuarantineBureauofthePeople's RepublicofChinaandWanhuaBiological EngineeringCo.Ltd.ofBeijingThis standardwas mainly draftedbyWanJicheng.Wan Zhijing,MiJiebo,Mo Jin,ZhengWenjie,TangXiaobin,Jian Liping.
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Fastdeterminationofcarbaryl,chlorpyrifos,propamocarb,metalaxyl,alachlor,metolachlorinfarmproducesbycolloidalgoldimmunoassay
1Scope
This standard specifies the method for the determination of carbaryl,chlorpyrifos,propamocarb,metalaxyl.alachlor.metolachlor inagricultureproductsbycolloidal gold immunoassay.This standard is applicableto the screen determination of carbaryl,chlorpyrifospropamocarb,metal-axyl.alachlor,metolachlorin vegetableand fruitsuchas cucumber,tomato,broccoli,cabbage.pome.Positive results should be confirmed by another method.2NormativereferencesbzxZ.net
The following documents are necessary for this standard.For dated references,only dated editionsshall apply to this standard. For undated references,the latest edition of the normative document(including subsequentamendments)referredtoapplies.GB/T6682Waterforanalytical laboratoryuse-Specificationandtestmethods3Principle
The sandwich enzyme-linked immunoassayand the competitiveenzyme-linked immunoassayare ap-plied to detect pesticide residues using large aperture membrane as a carrier and colloidal gold asmarkers of solid phase. Monocloneantibodies（sandwich imunoassay)or antigen （competitive im-munoassay) is coated in detection zone,and sheep anti-mouse polyclonal antibody is coated in thecontrol zone.Thereactionresult(colourribbon appearsonthetestpaper)will beobserved on thedetection zone and control zonebecause of the combination between antigen of sample and antibodymarked with colloidal gold.Then it canbe determined whetherthepesticide ismorethan MRLs basedonthephenomenon(numbersofcolourribbons).Reagentsandmaterials
Unlessotherwise specified,all the chemical reagents shouldbe analytically pure,\water\is thethree6
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gradewaterprescribedbyGB/T6682Sodiumchloride.
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4.2Colloidal gold immunoassay kit,The colloidal gold immunoassay kit made by Wanhua BiologicalEngineering Co.,Ltd. 1 Of Beijing is used in this standard. Different kit is used for different pesti-cide,soCarbarylKit、ChlorpyrifosKit、PropamocarbKit、MetalaxylKit、AlachlorKit andMetolachlorKit are applied. Every kit comprises positive reference,negative reference and test paper,which ispacking in alumiumfoilbag
Apparatus and equipment
Homogenizer.
Petri dish.
Samplepreparation
5 g (to 0. 1 g precision) of fruit or vegetable with the soil of surface wiped is cut into little piecesabout 1 cm2.After 0.5 g sodium chloride added,the mixture is put into the homogenizer and madeinto juice.Then stand for3min~5min,leave the liquid in thePetri dish fornext step.Determination
TheMAXendoftestpaper,whichistakenoutfromthealuminumfoil bagand reached roomtemper-ature in advance,is dipped into the sample extraction and the liquid level should not exceed MAXline.Theantibodylabeledwithcolloidalgoldwillmoveslowlytowardsanotherendofthetestpaperforthecapollary action result fromthemovementof sampleextraction solution.The result couldberead from the observation window in1min~5 min and the maximum time shouldn'tmore than5min.
8TheresultjudgementandexpressionThe result is judged as following described according to the colour ribbon in the detection and controlzone.
1) The company name involved in the standard method is only for reference and not related to any commercial aim,andthe analysts are encouraged touseproducts of different corporation ordifferent type.7
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Failure:Thereisno colour ribbon incontrol zone;Non-compliantresult：
Thecolour ribbonappears inbothdetectionzoneand control zone,duringtheprocessof detectionusing the sandwich enzyme-linked immunoassay test paper;A colour ribbon appears only in control zone.during the process of detection using the competitiveenzyme-linkedimmunoassaytestpaper;c)
Compliantresult：
A colour ribbon appears only in control zone,during the process of detection using the sandwich en-zyme-linkedimmunoassaytestpaper;The colour ribbon appears in both detection zone and control zone,during the process of detectionusing the competitive enzyme-linked immunoassay test paper;d)
Threshold value:If a colour ribbon appears in the control zoneand a vague ribbonappears in thedetection zone,during the process of detection using the competitive enzyme-linked immunoassaytestpaper.
Theresult judgementof detectionusingtestpaperseeannexA.The results showed positiveor threshold value of the sampleare“positivesuspicious\sample.Thesampleshould betestedformorethan twiceand confirmed byMS method.9
Limit of determination and specificity9.1
Limitof determination
Table 1—Limit of determinationName of peticide
Carbaryl
Chlorpyrifos
Propamocarb
Metalaxyl
Alachlor
Metolachlor
Corpgroup
Cabbage
Tomato.Cabbage,Broccoli
Spinage
Cucumber.grape
Cabbage
Cucumber.cabbage
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Limit of detection
Specificity
Non-compliantresultrateof positivesamples≥90%Compliantresultrateofblanksamples≥90%Falsecompliantrate5%
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AnnexA
(Informativeannex）
Theresultjudgementofdetectionusingtestpaper?）-- MAX
Non-compliant rcsult
Csnjli:un resull
Failure rusull
Figure A.1-The result judgement of detection using the sandwichenzyme-linkedimmunoassaytestpaperau
Complianr result
Culull resull
Kan coimplianl result
Failure Iesult
FigureA.2-Theresult judgementofdetectionusingthecompetitiveenzyme-linked immunoassaytestpaper2)
Non-commercial statement: The result judgement of detection using test paper in annex A are accomplished by theproducts of Wanhua Biological Engineering Co. ,Ltd. Of Beijing,the company name involved in the standard method isonly for reference and not related to any commercial aim,and the analysts are encouraged to use products of differentcorporation or different type.10
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