【商检行业标准(SN)】 进出口水果和蔬菜中阿维菌素残留量检测方法 液相色谱法

中华人民共和国出入境检验检疫行业标准SN/T2114—2008
进出口水果和蔬菜中阿维菌素残留量检测方法
液相色谱法
Determination of abamectin residues in fruit and vegetable forimportandexport-LCmethod
2008-09-04发布
中华人民共和国
国家质量监督检验检疫总局
2009-03-16实施
本标准的附录A、附录B为资料性附录前言
本标准由国家认证认可监督管理委员会提出并归口。本标准由中华人民共和国山东出入境检验检疫局负责起草。本标准起草人：李立、李金强、李佩暖本标准系首次发布的出人境检验检疫行业标准。iAoNirKAca-
SN/T2114—2008
1范围
进出口水果和蔬菜中阿维菌素残留量检测方法液相色谱法
本标准规定了水果及蔬菜中阿维菌素检测的制样和液相色谱检测方法SN/T2114—2008
本标准适用于苹果及波菜中阿维菌系残留量的检测。其他水果和蔬菜可以参照本标准2试样制备与保存
2.1试样制备
将所取样品缩分出1kg，取可食部分，经组织捣碎机捣碎，均分为两份，装入洁净容器内，作为试样密封并标明标记。
2.2试样保存
将试样于一18℃以下保存
在抽样和制样的操作过程中，应防止样品受到污染或发生残留物含量的变化3方法提要
试样中的阿维菌素用丙酮提取，经浓缩后，用SPEC1s柱净化，并用甲醇洗脱。洗脱液经浓缩、定容、过滤后，用配有紫外检测器的高效液相色谱测定，外标法定量4试剂和材料bZxz.net
除另有规定外，所有试剂均为分析纯，水为去离子水。4.1丙酮。
4.2甲醇：色谱纯。
4.3阿维菌素标准品：纯度≥96.0%。4.4阿维菌素标准储备液：称取0.1g（准确至0.0002g）阿维菌素标准品于100mL容量瓶中，用甲醇溶解并定容至刻度。配制成浓度为1.0mg/mL的标准储备液4.5阿维菌素标准工作液：根据需要移取适量的阿维菌素标准储备液，用甲醇稀释成适当浓度的标准工作液。标准工作液需每周配制一次。5仪器和设备
5.1高效液相色谱仪：配有紫外检测器。5.2组织捣碎机。
5.3振荡器。
5.4旋转蒸发器
5.5固相萃取柱：SPEC18规格：60mg/3mL；使用前用5mL甲醇和5mL水活化6测定步骤
6.1提取
称取试样约20g（精确至0.1g）于100mL具塞锥形瓶中，加人50mL丙酮，于振荡器上振荡0.5h1
SN/T21142008
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用布氏漏斗抽滤，用20mL×2丙酮洗涤锥形瓶及残渣。合并内酮提取液，于40℃水浴旋转蒸发至约2mL。
6.2净化
将上述的浓缩提取液完全转人SPECis柱，再用5mL水淋洗，去掉淋洗液。最后用5mL甲醇洗脱，收集洗脱液，用氮气吹至近干。准确加人1.0mL甲醇溶解残渣，用0.45um滤膜过滤，滤液供液相色谱测定。外标法定量。
6.3测定
6.3.1高效液相色谱条件：
色谱柱：ODS-Cls反相柱，4.6mmX125mm；a）：
b）流动相：甲醇：水（90+10)；c
流速：1.0mL/min：
d）检测波长：245nm；
柱温：40℃；
f）进样量：20μL。
6.3.2色谱测定
根据样液中阿维菌素含量情况，选定峰高相近的标准工作液，标准工作液和样液中阿维菌素响应值均应在仪器检测线性范围内·标准工作液和样液等体积参插进样。在上述色谱条件下，阿维菌素保留时间约为5.3min。
标准色谱图参见附录A.标准品紫外光谱图参见附录B。6.4空白试验
除不加试样外，均按照上述测定步骤进行。结果计算与表述
用色谱数据处理机，或按式（1)计算试样中阿维菌素残留量，计算结果需扣除空白值：h.c.v
式中：
X试样中阿维菌素残留量，单位为毫克每千克（mg/kg）；h
样液中阿维菌素峰高，单位为毫米（mm）；h
标准工作液中阿维菌素峰高，单位为毫米（mm）；标准工作液中阿维菌素浓度，单位为毫克每升（mg/L）；样液最终定容体积，单位为毫升（mL）；最终样液代表的试样量，单位为克（g）。测定低限和回收率
测定低限
本方法的测定低限为0.01mg/kg。8.2回收率
苹果样品中添加阿维菌素的浓度和回收率的实验数据：在0.01mg/kg时，回收率为82.5%；在0.05mg/kg时，回收率为87.5%；2
在0.50mg/kg时，回收率为95.0%。菠菜中添加阿维菌素的浓度和回收率的实验数据：——在0.01mg/kg时.回收率为83.0%；一在0.05mg/kg时，回收率为89.0%；—在0.50mg/kg时，回收率为97.0%SN/T2114—2008
SN/T 2114—2008
附录A
（资料性附录）
阿维菌素标准品液相色谱图
阿维菌素标准品液相色谱图
附录B
（资料性附录）
阿维菌素标准品紫外吸收光谱图250
阿维菌素标准紫外吸收光谱图
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Foreword
Annex A,Annex B of this standard are informative annexes.SN/T2114—2008
This standard was proposed by and is under the charge of the Certification and Accreditation of thepeople's Republic of China.This standard was drafted by Shandong Entey-Exit Inspection and Quar-antineBureauofthepeople'sRepublicofChina.The main drafters of this standard are LiLi,Li jinQiang.Li peinuanThis standard isa professional standard for inspectionand quarantine promulgated forthe first time.SN/T2114—2008
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DeterminationofabamectinresiduesinfruitanovegetableforimportandexportLCmethodScope
This standard specifies the methods of sample preparation and determination of abamectin residuesby highPerformance liquid chromatography in fruit and vegetableThis standard is applicable todetermination of abamectin residuce inappleand spinach.Samplepreparationandstorage
2. 1 Preparation of test sampleThe combined primary sanple is reduced to 1 kg,the edible portions are blended in a blender,and thendivided into two equal portions. Each portion is placed in a clean containeras the test sample,whichis then sealed and labeled.
2.2 Storage of test sample
Thetest sampleshouldbestored below-18℃In the course of sampling and sample preparation.precautions shall be taken to avoid contaminationor any factors which may cause the change of residue content3Principle
The abamectin residues in the test sample are extracted by acetone,and then concentrated andcleaned up by solid phase extraction Cia cartridge.Elute with methanol,and the eluate is concentrat-ed,made up to volumeand filtered.Determination is made by highperformance liquid chromatogra-phy equipped with UV detector,using external standard method.4
Reagents andmaterials
Unless otherwise specified.the reagents should be analytically pure,\water\is distilled water.6
Methanol:HPLC grade.
Abamectinstandard:Purity96.0%SN/T 2114—2008
Abamectin standard stock solution:Weigh 0.1 g (accurate to 0.0002 g)of abamectin standard4.4
intoa 1oo mL volumetric flask,dissolve with methanol,mixthroughly.The concentration of thestandardstocksolutionis1.omg/mL4.5
Abamectin standard working solution:According tothe requirement prepare standard workingsolution of appropriate concentration by diluting the standard stock solution with methanol. Preparethestandardworkingsolutionfleshlyeveryweek5
Apparatusand equipment
Highperformanceliquidchromatograph.equippedwithUVdetectorHigh-speed blender.
Rotaryevaporator
Cleanup column:SPECiacartridge,60 mg/3mL activated with5 mL of methanoland rinsedwith5mL of water.
Procedure
6.1 Extraction
Weight ca20g(accurateto0.1g)intoa100mL conical flaskwith stopper.add50 mLof acetone.shake for0.5hwithshaker,the extract isfilteredbysuctionthroughabuchnerfunnel,washtheresi-due and the flaskwith2x20o mLacetone,combine acetoneand evaporated to ca2mL with rotary e-vaporator under 40 ℃.
6.2 Cleanup
Transfer the concentrated extract into a SPE Cis cartridge,the effluent is discarded and rinsing with5 mL of water.Then elute with 5 mL of methanol.collect the eluate and nearly to dryness with astream of nitrogen.Make up to 1. O mL with methanol.Filter the solution with the filter membrane ofSN/T2114—2008
o.45μm.ThefiltrateisusedforHPLCdetermination.6.3Determination
6.3.1HPLCoperatingcondition：a)
HPLcolumn:ODSCig.4.6mmx125mm：Mobile phase:Methanol :water(90+10);Flow rate:1.0 mL/min
Wavelength:245nm
Columntemperature：4o℃；
Injection volume:20 μL.
HPLCdetermination
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According to the approximate concentration of abamectin in the sample solution,select the standardworking solutionwithsimilarpeakheighttothat of thesamplesolution.The responsesof abamectininthestandard working solution and sample solution shouldbe in the linear range of the instrumentaldetection. The standard solution should be injected in-between the injections of the sample solutionof equal volume. Under the above operating condition,the retention time of abamectin is 5.3 min.ForHPLCchromatogramofthestandard,seefigureinannexAandforphotodiodearrayspectrogramof the standard,see figue in annex B.6.4 Blank test
The operation of the blank test is the same as that described in the method of determination, butwithoutadditionof sample
Calculationand expression of the resultCalculate the content of abamectin residues in the test sampleby dateprocessororaccordingto theformula(1),theblankvolueshoulebesubtractedfromtheaboveresultofcalculationX=
where：
X-theresiduecontentofabamectininthetestsample,mg/kg;8
hthepeakheightofabamectininthesamplesolution.mm;hthepeak heightof abamectin inthe standard solution.mm;ctheconcentrationof abametion in the standard working solution,uμg/mL;V-thefinalvolumeofthesamplesolution.mLm-thecorrespondingmassofthetestsampleinthefinalsamplesolution,g8Limit of determination and recovery8.1Limit of determination
Thelimitofdeterminationofthismethodiso.o1mg/kg8.2Recovery
SN/T2114—2008
According to theexperimental data,the fortifying concentrations of abamectin in apple and its corre-spondingrecoveriesare：
—0.01mg/kg,the recovery82.5%:0.05mg/kg,therecovery87.5%;
-0.50 mg/kg.therecovery95.0%The fortifying concentrations of abamectin in spinach and its corresponding recoveriesare:0.01mg/kg,the recovery83.0%;0.05 mg/kg,the recovery 89.0%;0.50 mg/kg,the recovery 97.0%o
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AnnexA
(informative)
Chromatogramof thestandard
Figure A. 1-Liquid chromatogram of abamectin standardAnnexB
(informative)
spectrogram of thestandard
-SpectrogramofabamectinstandardFigureB.1-
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