SN∕T 1071.1-2019 出口粮谷中环庚草醚残留量的测定.pdf

中华人民共和国出入境检验检疫行业标准SN/T 1071.120192019-12-27 发布2020-07-01 实施中 华 人 民 共 和 国 海 关 总 署发 布ICS 67.040CCS C 53Determination of cinmethylin residue in cereals for export出口粮谷中环庚草醚残留量的测定出口粮谷中环庚草醚残留量的测定.indd 12020/1/16 20:23:55ISN/T 1071.12019前 言本标准依据 GB/T 1.12009标准化工作导则 第 1 部分：标准的结构和编写给出的规则起草。请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别这些专利的责任。本标准代替 SN/T 1017.12001出口粮谷中环庚草醚残留量检验方法。本标准与 SN/T 1017.12001 的主要差异如下：修改标准名称；扩大标准适用范围至糙米、大米、大麦、小麦、玉米和绿豆；增加气相色谱-质谱/质谱法。本标准由中华人民共和国海关总署提出并归口。本标准起草单位：中华人民共和国杭州海关。本标准主要起草人：黄超群、谢文、李翼、楼成杰、童赟恺、蒋沁婷、陈丽。本标准所代替标准的历次版本发布情况为：SN/T 1017.12001。出口粮谷中环庚草醚残留量的测定.indd 12020/1/16 20:23:551SN/T 1071.12019出口粮谷中环庚草醚残留量的测定1 范围本标准规定了出口粮谷中环庚草醚残留量测定的气相色谱-质谱和气相色谱-质谱/质谱检测方法。本标准适用于出口糙米、大米、大麦、小麦、玉米、绿豆中环庚草醚残留量的测定。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，仅所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/T 6682 分析实验室用水规格和试验方法第一法 气相色谱-质谱法3方法提要试样中环庚草醚经丙酮、水提取，用正己烷抽提，经弗罗里硅土柱净化后，气相色谱-质谱法测定，外标法定量。4试剂和材料除特殊注明外，所用试剂均为色谱纯，所用水为符合 GB/T 6682 规定的一级水。4.1丙酮。4.2正己烷。4.3无水乙醚。4.4氯化钠：分析纯。4.5无水硫酸钠：分析纯，650 灼烧 4 h。4.6正己烷-乙醚（9+1，V/V）：取 900 mL 正己烷和 100 mL 无水乙醚混合均匀。4.7氯化钠溶液（50 g/L）：称取 50 g 氯化钠用水溶解并定容至 1 000 mL，混匀。4.8标准物质：环庚草醚，C18H26O2，CAS 登录号 87818-31-3，浓度 10 g/mL，20 避光保存。4.9标准工作溶液：根据需要，将标准物质（4.8）用正己烷稀释成适当浓度的标准工作溶液，0 4 保存。4.10弗罗里硅土柱：270 mm 10 mm（内径）玻璃柱，底部垫有脱脂棉和约 10 mm 高无水硫酸钠，填料为 1.00 g 弗罗里硅土（经 650 灼烧 4 个小时），上层再覆盖约 10 mm 高无水硫酸钠。4.11无水硫酸钠柱：75 mm30 mm（内径），内装 30 mm 高无水硫酸钠。出口粮谷中环庚草醚残留量的测定.indd 12020/1/16 20:23:552SN/T 1071.120195仪器和设备5.1气相色谱-质谱仪：配有 EI 源。5.2分析天平：感量 0.000 1 g 和 0.01 g。5.3涡旋混合器。5.4振荡器。5.5旋转蒸发仪。6试样制备与保存6.1试样制备取代表性样品糙米、大米、玉米、大麦、小麦、绿豆各约 500 g，用粉碎机粉碎，混匀，装入洁净的盛样容器中，密封并标明标记。6.2试样保存试样于 0 4 保存。在制样的操作过程中，应防止样品受到污染或发生残留物含量的变化。7测定步骤7.1提取准确称取 10 g 试样（精确至 0.01 g），置于 100 mL 三角烧瓶中，加入 15 mL 水，浸泡 2 h，再加入 15 mL 丙酮，振荡 30 min，过滤。残渣用 15 mL 丙酮再提取一次，合并滤液于 250 mL 分液漏斗中，加入 5 mL 氯化钠溶液，用 20 mL 正己烷提取。水层再用 20 mL 正己烷提取一次，合并正己烷层。正己烷层过无水硫酸钠柱（4.11）脱水，浓缩至干，待净化。7.2净化准确移取 10 mL 正己烷溶解残渣。取 1.0 mL，转移至弗罗里硅土柱（4.10）内，加 10 mL 正己烷淋洗，弃去流出液。用 10 mL 正己烷-乙醚混合溶液（4.6）洗脱，收集洗脱液，浓缩至干，加 1.0 mL正己烷，涡旋，供气相色谱-质谱仪测定。7.3测定7.3.1气相色谱-质谱条件气相色谱-质谱条件如下：a）色谱柱：HP-5MS，30 m0.25 mm0.25 m，或相当者；b）柱温：60 保持 1 min，以 30 /min 的速率升温至 180，随后以 10 /min 的速率升温至 250，保持 5 min；c）进样口温度：250 ；d）载气：氦气，纯度 99.999%，流量 1.0 mL/min；e）进样量：2 L；f）进样方式：不分流进样；g）电离方式：EI；h）电离能量：70 ev；出口粮谷中环庚草醚残留量的测定.indd 22020/1/16 20:23:553SN/T 1071.12019i）离子源温度：230 ；j）四极杆温度：150 ；k）接口温度：280 ；l）检测方式：选择离子监测；m）监测离子：105*、123、169；an）溶剂延迟：7 min。7.3.2定量测定按 7.3.1 仪器条件测定标准工作溶液和样液，以外标曲线法计算样液中的环庚草醚含量。如果样液中环庚草醚的含量超出标准曲线范围，应用正己烷稀释后再进行分析。在上述条件下，环庚草醚的参考保留时间约为 9.5 min。标准溶液选择离子监测色谱图参见附录 A 中图 A.1。7.3.3定性测定按照上述仪器条件测定样液和标准工作溶液，如果样液与标准工作液中待测物质色谱峰相对保留时间偏差在 0.5范围内；定性离子对的相对丰度与浓度相当的标准工作溶液的相对丰度一致，相对丰度允许误差不超过表 1 规定的范围，则可判断样品中存在相应的被测物。表 1 相对离子丰度最大容许误差相对离子丰度（基峰）%50 2050 1020 10最大容许误差%101520507.4空白试验除不加试样外，均按上述操作步骤进行。8结果计算和表述用色谱数据处理机或按公式（1）计算试样中环庚草醚的残留量：1 0001 000c VXm=（1）式中：X 试样中环庚草醚的残留量，单位为毫克每千克（mg/kg）；c 从标准曲线上得到的待测物质的溶液浓度，单位为微克每毫升（g/mL）；V 样液最终定容体积，单位为毫升（mL）；m 最终样液所代表的试样质量，单位为克（g）；计算结果应扣除空白值。9定量限和回收率9.1定量限本方法对于糙米、大米、大麦、小麦、玉米和绿豆中环庚草醚的定量限均为 0.01 mg/kg。*定量离子出口粮谷中环庚草醚残留量的测定.indd 32020/1/16 20:23:554SN/T 1071.120199.2回收率本方法回收率试验数据见表 2。表 2 回收率试验数据（n=6）添加浓度mg/kg回收率范围/%糙米大米大麦小麦玉米绿豆0.017286738583987191789278980.0282918610387110749085100861030.10891028210485101821017789851040.207690859784997997879989101第二法 气相色谱-质谱/质谱法10方法提要试样中环庚草醚经丙酮、水提取，用正己烷抽提，经弗罗里硅土柱净化后，气相色谱-质谱/质谱法测定，外标法定量。11试剂和材料除特殊注明外，所用试剂同 4.14.6 和 4.84.9。11.1弗罗里硅土柱：6 mL，1 g。11.2微孔滤膜：0.45 m，有机相。12仪器和设备除特殊注明外，所用仪器和设备同 5.25.5。12.1气相色谱-质谱/质谱仪：配有 EI 源。12.2离心机：最大转速 4 000 r/min。12.3固相萃取仪。13试样制备与保存试样制备与保存同 6.1 和 6.2。14测定步骤14.1提取准确称取 10 g 试样（精确至 0.01 g），置于 50 mL 离心管中，加入 10 mL 水，浸泡 2 h，再加入出口粮谷中环庚草醚残留量的测定.indd 42020/1/16 20:23:555SN/T 1071.1201910 mL 丙酮，振荡 10 min，4 000 r/min 离心 3 min，收集上清液于另一 50 mL 离心管中。残渣用 20 mL 丙酮再提取一次，合并上清液，加入约 4 g 氯化钠和 15 mL 正己烷，涡旋 1 min，离心，上层溶液转移至 100 mL 浓缩瓶中，下层溶液再加入 15 mL 正己烷，涡旋 1 min，离心，合并正己烷层，浓缩至近干，待净化。14.2净化准确移取 10 mL 正己烷溶解残渣。取 1.0 mL，转移至弗罗里硅土柱（11.1）内，加 10 mL 正己烷淋洗，弃去流出液。用 10 mL 正己烷-乙醚混合溶液（4.6）洗脱，收集洗脱液，浓缩至干，加 1.0 mL正己烷，涡旋，经滤膜过滤，供气相色谱-质谱/质谱仪测定。14.3测定14.3.1气相色谱-质谱/质谱条件气相色谱-质谱/质谱条件如下：a）色谱柱：HP-5MS，30 m0.25 mm0.25 m，或相当者；b）柱温：60 保持 1 min，以 30 /min 的速率升温至 180，随后以 10 /min 的速率升温至 250，保持 5 min；c）进样口温度：250 ；d）载气：氦气，纯度 99.999%，流量 1.0 mL/min；e）进样量：2 L；f）进样方式：不分流进样；g）电离方式：EI；h）电离能量：70 ev；i）离子源温度：300 ；j）接口温度：280 ；k）检测方式：选择反应监测；l）监测离子对信息见表 3；m）溶剂延迟：7 min。表 3 选择反应监测离子对及碰撞能量化合物母离子 m/z子离子 m/z碰撞能量（V）环庚草醚169.0107.1*10123.210 注：带*为定量离子14.3.2定量测定按 14.3.1 仪器条件测定标准工作溶液和样液，以外标曲线法计算样液中的环庚草醚含量。如果样液中环庚草醚的含量超出标准曲线范围，应用正己烷稀释后再进行分析。在上述条件下，环庚草醚的参考保留时间约为 9.4 min。标准溶液选择反应监测色谱图参见附录 A 中图 A.2。14.3.3定性测定按照上述仪器条件测定样液和标准工作溶液，如果样液与标准工作液中待测物质色谱峰相对保留时间偏差在 0.5范围内；定性离子对的相对丰度与浓度相当的标准工作溶液的相对丰度一致，出口粮谷中环庚草醚残留量的测定.indd 52020/1/16 20:23:556SN/T 1071.12019相对丰度允许误差不超过表 4 规定的范围，则可判断样品中存在相应的被测物。表 4 相对离子丰度最大容许误差相对离子丰度（基峰）%50 2050 1020 10最大容许误差%2025305014.4空白试验除不加试样外，均按上述操作步骤进行。15结果计算和表述结果计算和表述同 8。16定量限和回收率16.1定量限本方法对于糙米、大米、大麦、小麦、玉米和绿豆中环庚草醚的定量限均为 0.01 mg/kg。16.2回收率本方法回收率试验数据见表 5。表 5 回收率试验数据（n=6）添加浓度mg/kg回收率范围/%糙米大米大麦小麦玉米绿豆0.017993769884988194859781970.0277967688869983948794871020.10811007993779082968698851010.208796789287968310382958698出口粮谷中环庚草醚残留量的测定.indd 62020/1/16 20:23:557SN/T 1071.12019附 录 A（资料性附录）气相色谱-质谱法和气相色谱-质谱/质谱法标准溶液色谱图图 A.1 环庚草醚标准溶液的选择离子监测（SIM）色谱图（0.01 g/mL）图 A.2 环庚草醚标准溶液的选择反应监测（SRM）色谱图（0.01 g/mL）出口粮谷中环庚草醚残留量的测定.indd 72020/1/16 20:23:569SN/T 1071.12019ForewordThis standard was drafted in accordance with the GB/T 1.12009Directives for standardization-part 1：structure and drafting of standards.This standard is the revision of SN/T 1017.12001Method for the determination of cinmethylin residues in cereals for export.The differences between this standard and SN/T 1017.12001 are as follows：Changed the name of the standard；Extended the scope to the matrices of unpolished rice，rice，barley，wheat，corn and mung bean；Added LC-MS/MS method.This standard is proposed by and is under the charge of General Administration of Customs of the Peoples Republic of China.This standard was drafted by Hangzhou Customs District of the Peoples Republic of China.The main drafters of this standard are Huang Chaoqun，Xie Wen，Li Yi，Lou Chengjie，Tong Yunkai，Jiang Qinting，Chen Li.This standard replaces the standard previous published as：SN/T 1017.12001.出口粮谷中环庚草醚残留量的测定.indd 92020/1/16 20:23:5611SN/T 1071.12019Method for the determination of cinmethylin residue in cereals for export1 ScopeThis standard specifies the determination of cinmethylin in cereals for export by gas chromatography-mass spectrometry and gas chromatography-tandem mass spectrometry.This standard is applicable to the determination of cinmethylin in unpolished rice，rice，barley，wheat，corn and mung bean for export.2 Normative referencesThe following documents is necessary for this standard.For dated references，only dated editions shall apply to this standard.For undated references，the latest edition of the normative document referred to applies.GB/T 6682 Water for analytical laboratory usespecification and test methodsMethod GC-MS Method3 PrincipleCinmethylin in the test samples is extracted with water-acetone.The extract is partitioned with hexane，cleaned up by florisil solid phase and detected by gas chromatography-mass spectrometry using external standard method.4 Reagents and materialsUnless otherwise specified，all the reagents used should be chromatographic grade，“water”is the first grade water prescribed by GB/T 6682.4.1 Acetone.4.2 Hexane.4.3 Diethyl ether.4.4 Sodium chloride：analytical grade.4.5 Anhydrous sodium sulfate：analytical grade，Ignite at 650 for 4h.4.6 Hexane-diethyl ether（9+1，V/V）：Volume 900 mL hexane，then add 100 mL diethyl ether，mix them.4.7 Sodium chloride solution（50 g/L）：dissolve 50 g of sodium chloride in 1 000 mL of water.4.8 Standard：Cinmethylin，C18H26O2，CAS NO.87818-31-3，concentration of 10 g/mL，stored under 20 in the dark.4.9 Standard work solution：According to the requirement，diluted the standard（4.8）with hexane.Stored under 0 4.出口粮谷中环庚草醚残留量的测定.indd 112020/1/16 20:23:5612SN/T 1071.120194.10 Florisil column：grass column of 270 mm10 mm（i.d），absorbent cotton and 10 mm of anhydrous sodium sulfate at the bottom，filling with 1 g of florisil（Ignite at 650 for 4h），the upper layer was covered with 10 mm of anhydrous sodium sulfate.4.11 Anhydrous sodium sulfate column：75 mm35 mm（i.d），filled with 30 mm of anhydrous sodium sulfate.5 Apparatus and equipment5.1 Gas chromatography-mass spectrometer，equipped with EI source.5.2 Electronic balance：accurate to 0.000 1 g and 0.01 g.5.3 Vortex mixer.5.4 Oscillator.5.5 Rotary vacuum evaporator.6 Preparation and storage of test sample6.1 Preparation of test sampleTake approximately 500 g of representative unpolished rice，rice，barley，wheat，corn and mung bean，smashed thoroughly by a chopper，mixed thoroughly and put into clean containers，which is sealed and labeled.6.2 Storage of test sampleTest samples should be stored at a temperature ranged from 0 4.In course of sampling and sample preparation，attention must be taken to avoid contamination or any factors which may cause the change of residue content.7 Procedure7.1 ExtractionWeigh 10 g（accurate to 0.01 g）of the test samole into a 100 mL concial flask.Add 15 mL of water，soak for 2h，add 15 mL of acetone and shake for 30 min，then filter.Extract the residues once more with 15 mL of acetone.Combine the filtrates in a 250 mL separatory funnel.Add 5 mL of sodium chloride solution，and then extract with 20 mL and 20 mL of hexane respectively.Combine the hexane layers and pass through the column of anhydrous sodium sulfate，then condensed to dry，waiting for cleaning-up.7.2 Cleaning-upAdd 10 mL of hexane to dissolve the residue.1.0 mL of the solution was transfered to a florisil column（4.10）.After sample loading，the column was washed with 10 mL of hexane，and finally cinmethylin was eluted by 10 mL of hexane-diethyl ether（4.6）.The eluate was condensed to dry.Dissolve the residue with 1.0 mL of hexane，and detect by GC-MS.出口粮谷中环庚草醚残留量的测定.indd 122020/1/16 20:23:5613SN/T 1071.120197.3 Determination7.3.1 GC-MS operating conditionGC-MS operating condition is as follows：a）Chromatographic Column：HP-5MS，30 m 0.25 mm（i.d），film thickness 0.25 m，or equivalent；b）Column temperature：60（1 min）30/min 180 10/min 250（5 min）；c）Injection port temperature：250；d）Carrier gas：Helium，purity 99.999%，flow rate 1.0 mL/min；e）Injection volume：2.0 L；f）Injection mode：splitless；g）Electron ionization mode：EI；h）Electron ionization energy：70 eV；i）Ion source temperature：230 ；j）Quadrupole temperature：150 ；k）Interface temperature：280 ；l）Scan mode：SIM；m）Selected monitoring ion（m/z）：Quantitation 105，Qualification 123，169；n）Solvent delay：7 min.7.3.2 Quantitative determinationAnalyze standard working solutions and samples with external standard menthod using conditions established in sec.7.3.1.If the concentration of cinmethylin exceeds the linear range of the calibration curve，the final solution should be diluted with hexane and reanalyzed.Under the above chromatographic conditions，the retention time of chromatographic peak of cinmethylin is about 9.5 min.For the GC-MS SIM chromatogram refer to fig.A.1 in Appendix A.7.3.3 Qualitative determinationThe accordance between the retention time of the test sample solution and the time of the standard，the consistency between the abundance ration of the selected ions from the sample and the ratio of ions from standard，and the similarity subject to the allowance（see table 1），can deliver the positive judgment of analyte detection.Table 1 Maximum allowance of relative ion abundanceRelative ion abundance（base peak）%50 2050 1020 10Maximum allowance%101520507.4 Blank testThe operation of the blank test is the same as that described in the method of determination，but with omission of sample addition.8 Calculation and expression of the resultCalculate the content of cinmethylin in the test sample by data processor or according to the following formula：出口粮谷中环庚草醚残留量的测定.indd 132020/1/16 20:23:5614SN/T 1071.120191 0001 000c VXm=（1）Where：X the content of butamifos in the test sample，mg/kg；C the concentration of butamifos in sample solution calculated by standard curve，g/mL；V the final volume of the sample solution，mL；m the corresponding mass of the test sample in the final sample solution，g.The result should be deducted by value of blank test.9 Limit of quantitation and recovery9.1 Limit of quantitationThe limits of quantitation for cinmethylin in unpolished rice，rice，barley，wheat，corn and mung bean are 0.01 mg/kg.9.2 RecoveryThe results of recovery test are listed in Table 2.Table 2 Results of recovery test（n=6）Spike levelmg/kgRecovery range%unpolished ricericebarleywheatcornmung bean0.017286738583987191789278980.0282918610387110749085100861030.10891028210485101821017789851040.207690859784997997879989101Method GC-MS/MS Method10 PrincipleCinmethylin in the test samples is extracted with water-acetone.The extract is partitioned with hexane，cleaned up by florisil solid phase and detected by gas chromatography-tandem mass spectrometry using external standard method.11 Reagents and materialsUnless otherwise specified，reagents are the same as section 4.1 to 4.6 and section 4.8 to 4.9.11.1 Florisil SPE column：6 mL，1 g.11.2 Millipore filter：0.45 m，organic phase.出口粮谷中环庚草醚残留量的测定.indd 142020/1/16 20:23:5615SN/T 1071.1201912 Apparatus and equipmentUnless otherwise specified，apparatus are the same as section 5.2 to 5.5.12.1 Gas chromatography-tandem mass spectrometry，equipped with EI.12.2 Centrifuge（4 000 r/min）.12.3 Solid Phase Extraction System.13 Preparation and storage of test samplePreparation and storage of test samples are the same as section 6.1 and 6.2.14 Procedure14.1 ExtractionWeigh 10 g（accurate to 0.01 g）of the test samole into a 50 mL polypropylene tube，10 mL of water was added.The sampled was soaked for 2 h.Then 10 mL of acetone was added.After violent shaking for 10 minutes，the sample was centrifuged at 4 000 r/min for 3 minutes.The upper solution was transferred into another 50 mL polypropylene tube，and the residue was extracted with 20 mL of acetone once more.Combine the solution in the same tube.4 g sodium chloride and 15 mL of hexane were added.The mixture was been vortex for 1 minute，centrifuged at 4 000 r/min for 3 minutes.The upper solution was transferred into a concentrate bottle，and the residue was extracted with 15 mL of hexane once more.Combine the layer of hexane，condensed to nearly dry，waiting for cleaning-up.14.2 Cleaning-upAdd 10 mL of hexane to dissolve the residue.1.0 mL of the solution was transfered to a florisil column（11.1）.After sample loading，the spe column was washed with 10 mL of hexane，and finally cinmethylin was eluted by 10 mL of hexane-diethyl ether（4.6）.The eluate was condensed to dry.Dissolve the residue with 1.0 mL of hexane and detect by GC-MS/MS.14.3 Determination14.3.1 GC-MS/MS operating conditionGC-MS/MS operating condition is as follows：a）Chromatographic Column：HP-5MS，30 m 0.25 mm（i.d），film thickness 0.25 m，or equivalent；b）Column temperature：60（1 min）30/min 180 10/min 250（5 min）；c）Injection port temperature：250；d）Carrier gas：Helium，purity 99.999%，flow rate 1.0 mL/min；e）Injection volume：2.0 L；f）Injection mode：splitless；g）Electron