1、中华人民共和国出入境检验检疫行业标准SN/T 1591一2005进出口茶叶中9种有机杂环类农药残留量的检验方法2005-05-20发布Lnspection of 9 organic heterocyclic pesticides in tea for import and export 2005-12-01实施中华人民共和国发布国家质量监督检验检疫总局SN/T 1591一2005前士一同本标准的附录A为资料性附录。本标准由国家质量监督检验检疫总局提出井归口。本标准由中华人民共和国湖南出入境检验检疫局负责起草。本标准主要起草人:李拥军、黄志强、戴华、张莹。本标准系首次发布的出入境检验检疫行业标准
2、。I 1 范围进出口茶叶中9种有机杂环类农药残留量的检验方法SN/T 1591-一2005本标准规定了进出口茶叶中9种有机杂环类农药残留量检验的抽样和制样、测定方法、测定低限及回收率。本标准适用于进出口茶叶中芳去津、乙烯菌利核、腐霉利、氟菌略、抑霉略、唾嗦酣、丙环略、氯苯喃院醇、贴瞒灵残留量的检验。2 抽样和制样2. 1 检验批以不超过2000件为一检验批，同一检验批内商品应具有相同的特征，如包装、标记、产地、规格、等级等。2.2 抽样数量表1抽样数量表单位为件批量最低抽样数15 1 650 2 51500 11 5011 000 16 1 0011 500 17 1 5012 000 18
3、2.3 抽样工具a) 取样勺。b) 分样布:塑料布。c) 分样板。d) 盛样器:筒或塑料袋，可密封。2.4 抽样方法按2.2规定的抽样件数从货品堆装的不同部位随机抽取样箱，逐件开启。打开箱盖，将箱内茶叶全部倒入塑料布上，用取样勺在各部位抽取茶样，每件抽取的量约500g.作为原始样。混合原始样品的总量不少于2kg。将所取全部原始样品充分拌匀，用四分法(或用分样器)缩分出约500g.立即装入清洁、干燥的样品筒内密封，并标明标记，及时送实验室。2.5 试样制备将所取回的样品全部磨碎，使通过孔径为0.84mm筛。?昆匀，均分成两份，装入清洁的容器内，作为试样。密封，并标明标记。2.6 试样保存将试样于
4、一50C以下避光保存。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。SN/T 1591-2005 3 测定方法3. 1 方法提要试样中残留的劳去津、乙烯菌利核、腐霉利、氟菌瞠、抑霉略、唾嗦酣、丙环瞠、氯苯喃睫醇、贴瞒灵用丙酣正己皖提取。采用活性炭小柱和中性氧化铝小柱净化，被测物用丙酬-正己烧洗脱。净化后用气相色谱仪，配质谱检测器测定，外标法定量。3.2 试剂和材料除特殊规定外，所有试剂均为分析纯，水为蒸锢水。3.2. 1 丙酣:重蒸锢。3.2.2 正己炕z重蒸馆。3.2.3 氯化铀。3.2.4 元水硫酸铀:经650.C灼烧4h，置干燥器中备用。3.2.5 活性炭固相萃取小柱
5、:250mg或相当者。3.2.6 中性氧化铝固相萃取小柱:250mg或相当者。3.2.7 丙酬正己皖(1+3)溶液。3.2.8 丙酬-正己烧(2+1)榕液。3.2.9 农药标准品:纯度二三98%。3.2. 10 标准溶液z准确称取适量的芳去津、乙烯菌利核、腐霉利、氟菌瞠、抑霉略、唾嗦酣、丙环瞠、氯苯蠕睫醇、贴瞒灵标准品，用丙酣配制成1.00 mg/mL标准储备液，再根据需要用正己烧稀释成相应的标准工作溶液。3.3 仪器和设备a) 气相色谱仪，配质量选择性检测器;b) 固相萃取装置，带真空泵;c) 多功能微量化样品处理仪，或相当者;d) 离心机:4000r/min; e) 涡旋棍匀器;f) 离心
6、管:15mL; g) 刻度试管:15mL; h) 微量注射器:10L。3.4 测定步骤3.4. 1 提取准确称取1g均匀试样(精确至0.001g)于15mL离心管中，加入1g氯化铀(3.2.3)，加入2mL 蒸榴水，于1昆匀器上混匀30s，放置30min，再加入4mL丙酣和正己烧混合液(3.2.7)，在混匀器上混匀2min.在2500 r/min下离心1min，吸取上层正己炕提取液于另一15mL刻度试管中。再分别加入2mL丙酣和正己烧1昆合液(3.2.7)重复提取两次，合并提取液，加入1g元水硫酸铀干燥。将干燥后的提取液完全转移至另一干净刻度试管中置于微量化样品处理仪上在50.C，氮气流吹至约
7、1mL(溶液A)。3.4.2 净化将活性炭固相萃取小柱和中性氧化铝固相萃取小柱(活性炭固相萃取小柱内填约1cm高的无水硫酸铀层)自上而下安装在固相萃取的真空抽滤装置上，先用1mLX3丙酣预淋洗小柱，再用1mLX3正己皖预淋洗小柱，保持滴速0.5mL/min，弃去所有淋洗液。将榕液A依次过活性炭固相萃取小柱和中性氧化铝固相萃取小柱，再用6.0mL丙酣和正己烧混合液(3.2.8)淋洗柱子，收集全部洗脱液，置于50.C下，氮气流吹至近干。最后用正己烧定容于0.5mL , 供GC-MSD分析备用。3.4.3 测定3.4.3.1 气相色谱-质谱条件SN/T 1591-2005 a) 色谱柱:石英毛细管柱
8、，5%苯基甲基聚硅氧炕固定相。30m X O. 20 mm (内径)，膜厚0.25m，或相当者;b) 色谱柱温度:700C保持2min，以80C/min上升至180oC，再以30C/min上升至280oC，保持农18 min; c) 进样口瘟度:2500C ; d) 色谱-质谱接口温度:220oC;e) 载气:氮气(纯度99.995%),0.6 mL/min; f) 进样量:1L; g) 进样方式:无分流进样，1min后开阀;h) 电离方式:EI;i) 电离能量:70 ev; j) 测定方式:选择离子监测方式CSIM); k) 监测离子Cm/z):见表2; 1) 榕剂延迟:15 mino 表2
9、9种杂环类农药的监测离子药采集时间/min监测离子(m/z)农药采集时间/min监测离子(m/z)努去津1520 173.187.200时.215唾嚓酣28. 230. 5 105时.172.175.305I 乙烯菌利核2025 187.198.212时.285丙环瞠30. 536 173) .191,259.261 腐霉利2526. 5 96) .255.283.285 氯苯电曾院醇3640. 5 139时.219.251.330氟菌瞠2526. 5 219.248.278时.287即主蜻灵40. 543 117.147) .309.364 抑霉嗖26.528.2 173时.215.240.
10、296a) 标记离子为定量离子。3.4.3.2 色情测定根据样液中莞去津、乙烯菌利核、腐霉利、氟菌瞠、抑霉瞠、唾嗦自同、丙环略、氯苯嘈院醇、贴蜻灵的含量情况，选定峰面积相近的标准工作榕液。标准工作溶液和样液中芳去津、乙烯菌利核、腐霉利、氟菌哇、抑霉略、唾嗦酣、丙环略、氯苯嘈院醇、贴瞒灵的响应值均应在仪器检测的线性范围内。对标准工作液和样液等体积参插进样测定。在上述色谱条件下，标准品SIM色谱图参见附录A中图A.1。3.4.3.3 质谱确证对标准榕液及样液均按3.4.3.1规定的条件进行测定，如果样掖中与标准榕液相同的保留时间有峰出现，则对其进行质谱确证。当待测物全部监测离子的相对丰度与标准品一
11、致，且相似度在土10%之内时，可确证此待测物。在上述气相色谱质谱条件下，9种杂环类农药的保留时间及监测离子丰度比Cm/z)见表30表39种杂环类农药的保留时间和监测离子丰度比农药保留时间/min监测离子丰度比秀去津18.41 173: 187: 200 215(26 : 3 : 100: 58) 乙烯商利核21. 09 187: 198: 212: 285(74: 89 : 100 86) 腐霉利25.46 96 : 255 : 283 : 285 (100 : 8 : 69 : 5) 3 SN/T 1591-2005 表3(续)农药保留时间/min监测离子丰度比氟菌瞠25. 67 219:
12、248: 278: 287 (18 : 7: 100: 53) 抑霉唾27.33 173: 215: 240: 296(76: 100: 9: 6) 喽嚓酣28. 30 105: 172: 175: 305 (100: 35: 25 : 6) 丙环R坐32.20 ,32.58 173: 191 : 259 : 261 (100: 27: 88: 57) 氯苯嗜睫醇39.12 139 : 219 : 251 : 330 (100 : 63 I 54 : 36) 盹瞒灵41. 64 117: l47: 309: 364 (15: 100: 7 I 6) 3.4.4 空白实验除不加试样外，均按上述测
13、定步骤进行。3.5 结果计算和表述用色谱数据处理机或按式(1)计算试样中芳去津、乙烯菌利核、腐霉利、氟菌略、抑霉略、睛菌略、喔嗦嗣、丙环唾、氯苯嘈院醇、盹蜻灵的含量:V一-m A一-A A一X . ( 1 ) 式中:X一-试样中秀去津、乙烯菌利核、腐霉利、氟菌瞠、抑霉睡、睛菌略、唾嗦酣、丙环睡、氯苯唔院醇、贴瞒灵的含量，单位为毫克每千克(mg/kg); A一-样液中莞去津、乙烯菌利核、腐霉利、氟菌睡、抑霉瞠、睛菌略、唾嗦酣、丙环瞠、氯苯嘈院醇、助蜡灵的峰面积，单位为平方毫米(mm2); Cs一-标准工作掖中芳去津、乙烯菌利核、腐霉利、氟菌睡、抑霉略、腊菌略、唾嗦酣、丙环睡、氯苯喃院醇、贴瞒灵的
14、浓度，单位为微克每毫升(g/mL); As -_标准工作液中芳去津、乙烯菌利核、腐霉利、氟菌瞠、抑霉瞠、腊菌略、唾嗦酣、丙环嗖、氯苯喃院醇、贴瞒灵的峰面积，单位为平方毫米(mm2);自V二一样液最终定容体积，单位为毫升(mL);m 最终样液所代表的试样量，单位为克(g)。注:计算结果须扣除空白值。4 测定低限、回收率4. 1 测定低限本方法各农药的测定低限分别是:芳去津:O. 02 mg/kg; 腐霉利:0.02mg/kg; 抑霉嗖:0.05mg/kg; 丙环瞠:O. 05 mg/kg; 贴瞒灵:O. 50 mg/kg; 乙烯菌利核:0.02mg/kg; 氟菌略:O. 38 mg/kg; 唾嗦
15、酣:0.01mg/kg; 氯苯嘈睫醇:0.02mg/kgo 4.2 回收率本方法各农药的添加浓度及回收率的实验数据z4 SN/T 1591-2005 亮去津:被度在O.02 mg/kg2. 0 mg/kg范围内，回收率为89.5%111. 4%; 乙烯菌利核:浓度在O.02 mg/kg2. 0 mg/kg范围内，回收率79.5%105. 4%; 腐霉利:浓度在O.02 mg/kg2. 0 mg/kg范围内，回收率为92.3%98. 5%; 氟菌瞠:浓度在O.38 mg/kg30. 0 mg/kg范围内，回收率为76.6%87. 9%; 抑霉瞠:浓度在O.05 mg/kg5. 0 mg/kg范围
16、内，回收率为72.6%85. 6%; 唾喋酣:被度在O.01 mg/kg1. 0 mg/kg范围内，回收率为93.5%103. 5%; 丙环唾:浓度在O.05 mg/kg5. 0 mg/kg范围内，回收率为81.1%108. 2%; 氯苯喃睫醇:浓度在O.02 mg/kg2. 0 mg/kg范围内，回收率为89.5%1l6. 2%; 贴瞒灵:浓度在O.25 mg/kg20. 0 mg/kg范围内，回收率为87.2%94. 8%。5 SN/T 1591-2005 Abundance 750000 700000 650000 18.41 600000 550000 500000 450000 21
17、. 09 350000 300000 250000 200000 150000 100000 50000 k 20.00 18.41 min-一秀去津;21. 09 min一乙烯菌利核;25.46 min腐霉利F25.67 min-一氟菌嗖;27.33 min-抑霉瞠p28.30 min-唾嚓酣;32.20 min,32. 58 min-一-丙环瞠;39.12 min-一氯苯略旋醇;41. 64 min 赂鳞灵。25.00 27.33 附景A(资料性附录)标准晶色谱圈28.30 32.58 32.20 30.00 35.00 39. 12 41. 64 40.00 固A.19种杂环类农药标准品
18、的SIM色谱固6 45.00 50.00 mm SN/T 1591-2005 Forword The annex A of this standard is an informative annex. This standard was proposed byand is under the charge of the State Administration for Ouality Super vision and Inspection and Ouarantine of the Peoples Republic of China. This standard was drafted by
19、Hunan Entry-Exit Inspection and Ouarantine Bureau of the People s Re public of China. The main drafters of this standard are Li Yongjun ,Huang Zhiqiang ,Dai Hua ,Zhang Ying. This standard is a professional standard promulgated for the first time. Note: This English version.a translation from the Chi
20、nese text. is solely for guidance. 7 SN/T 1591-2005 Inspection of 9 organic heterocyclic pestcides residues in tea for import and export 1 Scope This standard specifies the methods of sampling , sample preparation and determination by gas chro matography-mass spectrometry of 9 organic heterocyclic p
21、estcides resdues in tea for import and ex port. This standard is applicable to the determination of atrazine、vinclozolin、procmidone、triflumizole、imazalil、buprofezin、propiconazole、fenarimol、pyridabenresidues in tea for import and export. 2 Sampling and sample preparation 2.1 Inspection lot The quanti
22、ty of an inspection lot should not be more than 1 500 packages. The characteristics of the cargo within the same inspection lot , such as packing , mark, origin , specifi cation and grade , should be the same. 2.2 Quantity of sample taken Table 1 quantity of sample taken unlt:piece Number of pieces
23、in an each inspection lot Minimum number of pieces to be taken 1-5 6-50 2 51-500 11 501-1000 16 1 001-1 500 17 1 501-2000 18 2. 3 Sampling tools a) Sampling ladle. b) Sheet: Plastic sheet. c) Plate for quartering. d) Sample container:Can or plastic bag ,which can be sealed. 2.4 Sampling procedure Dr
24、awa number of sample packages specified in 2. 2 from different parts of the pile at random and open the packages one by one. Pour the tea from each packages on a clean plastic sheet and take up 8 SN/T 1591-2005 the sample from different parts of the out-poured tea as the primary sample. The amount t
25、aken from each package is ca 500 g , the total quantity of the primary samples should be at least 2 kg. Mix thor oughly the primary samples and reduce to ca 500 9 bquartering， promotly place in a clean , dry sam ple container, seal , label and sent to laboratory in time. 2. 5 Preparation of test sam
26、ples The mixed prima叩sampleis crushed with a grinder and let wholly pass through a O. 84 mm (pore size) sieve. Mix well and divided into two equal portions. Each portion is placed in a clean container as the test sample ,which is sealed and labeled. 2. 6 Storage of test samples The test samples shou
27、ld be stored below -5巳andkept away from light. In the course of sampling and sample preparation , precaution must be taken to avoid contamination or any factors which may cause the change of residue content. 3 Method of determination 3. 1 Principle 丁heatrazine、vinclozolin、procymidone，triflumizole、im
28、azalil、buprofezin、propiconazole、fenarimol、pyridaben residues in the test sample are extracted with acetone-n-hexane. cleaned up by passing through a active carbon and neutral aluminum oxide cartridge. The analyte is eluted with acetone-n hexane. Determination and confirmation made by means of gas ch
29、romatograph equipped with mass selective detector, using external standard method. 3. 2 Reagents and materials Unless otherwise specified ,the reagents should be analytically pure , Water is redistilled water. 3.2.1 Acetone: Redistill. 3.2.2 n-Hexane:Redistilled. 3. 2. 3 Sodium chloride. 3.2.4 Anhyd
30、rous sodium sulfate: Ignite at 650巳for4 h, and keep in a desiccator. 3.2.5 Active carbon cartridge:250 mg or equivalent. 3.2.6 Neutral aluminum oxide cartridge:250 mg or equivalent 3.2. 7 Acetone + n-hexane (1 + 3). 3.2.8 Acetone + n-hexane (2 + 1). 9 SNjT 1591-2005 3.2.9 Pesticides standard: Purity
31、注98%.3.2. 10 Standard solution preparation: Accurately weight an adequate amount of atrazine、vinclozolin、procymidone、triflumizole、imazalil、buprofezin、propiconazole、fenarimol、pyridabenstandards , dis solve in a small volume of acetone. Dilute with acetone to form a standard stock solution of 1. 00 mg
32、/mL in concerntration. Then dilute the standard stock solution with n-hexane to the required con cerntration as the standard working solution. 3.3 Apparatus and equipment a) Gas chromatograph ,equipped with mass selective detector. b) Solid phase extraction unit with mechanical vacuum pump. c) Multi
33、functional sample treatment unit for micro-chemical method, or equivalent. d) Vortex mixer. e) Centrifuge:4000 r/min. f) Centrifuge tube: 15 mL. g) Graduated tube: 15 mL. h) Mcro-syringe: 10L. 3. 4 Procedure 3. 4. 1 Extraction Weigh ca 1 9 (accurate to 0.001 g) of the test sample into a 15 mL Centri
34、fuge tube,add 1 9 sodium chloride and 2 mL redistilled water,blend for 30 s. stand for 30 min. Add 4 mL of acetone-n-hexane (1 +酌，blendfor 2 min in vortex mixer,centrifuge for 2 min under 2 500 r/min. Transfer the upper n-hexane extract into another 15 mL graduated tube,Repeat the procedure with 2 m
35、L of acetone-n hexane (1 + 3) twice, Combine acetone-n-hexane extracts, Add 1.0 9 anhdrous sodium sulfate to dry.丁hetotal extracts after dried was transferred to another clean graduated tube and evaporated to nearly 1 mL (solvent A) for clean with nitrogen stream at 50t on the multifunctiom:! sample
36、 tre时，ment unit for micro-chemical method. 3. 4. 2 Cleanup Set up the active carbon and neutral aluminum oxide cartridge from up to down on the solid phase extraction vaccum manifold (about 1 cm thickness anhydrous sodium sulfate was put into the active carbon cartridge). Then wash the active carbon
37、 and neutral aluminum oxide cartridge with 1 mL x 3 acetone and n-hexane respectively, keep flow speed at 0.5 mL/min. Discard the eluent. Transfer the solvent A into the cartridge. Wash the test tube with 6 mL acetone-n-hexane solution (2 + 1) and add into the cartridge. Collect the total eluent. an
38、d blow nearly dry with nitrogen stream at 500C ,Dissolve the residue and dilute exactlto O. 50 mL with n-hexane for GC/MSD. 3. 4. 3 Determination 3.4.3. 1 GC/MSD operating conditions 10 SN/T 1591-2005 a) Column: Fused quartz capillary column. (5%-phenyl)-methylolysiloxane.30 m X 0.20 mm (id) .film t
39、hickness 0.25m.or the equivalent; b) Column temperature:70t for 2 min.ramp at 80C/min to 1800C .ramp at 30C/min to 280oC. hold for 18 min; c) Injection port temperature:250t ; d) GC/MS interface temperature:220oC ; e) Carrier gas:Helium (purity99. 995%) .0. 6 mL/min; f) Injection volume: 1L; g) Inje
40、ction mode:Splitless.purge after 1 min; h) lonization mode:EI; i) lonization energ:70 ev; j) Acquistion mode:SIM; k) Monitor ion (m/z) :See table 2; 1) Sovlent delay: 15 min; 3. 4. 3. 2 GC/MSD determination Table 2 Monitor ion of 9 heterocyclic pestcides pestcides acquistion Monitor ion(m/z) pestcid
41、es acquistion time/min time/min atrazine 15-20 173，187，200时，215buprofezin 28.2-30.5 vinclozolin 20-25 187，198，212时，285propiconazole 30.5-36 procymidone 25-26.5 96时，255，283，285fenarimol 36-40.5 triflumizole 25-26.5 219，248，278时，287pyridaben 40.5-43 imazalil 26.5-28.2 173时，215，240，296a) are quantitati
42、ve ion. Monitor ion(m/z) 105时，172，175，305173剖，191，259，261139时，219，251，330117，147酣，309，364According to the approximate concentration of atrazine、vinclozolin、procymidone、triflumizole、imazalil、buprofezin、propiconazole、fenarimol、pyridabenin the test sample solution, select the standard working solution
43、with similar peak area to that of sample solution.丁heresponses of atrazine、vinclozolin、procymidone、triflumizole、imazalil、buprofezin、propiconazole、fenarimol、pridabenin the stand ard working solution and sample solution should be in the linear range of the instrumental detection. The standard working
44、solution should be injected randomly in between the injections of sample solu tion of equal volume. Under the above GC/MSD conditions, the retention time of 9 heterocyclic pesti cides standard see to table 2. For SIM chromatogram of 9 heterocclic pesticides standard ,See Fig. A 1 in annex A. 3.4.3.3
45、 GC/MSD confirmation According to the operating condition assigned in 3. 4. 3. 1 analyze the standard solution and sample solution. If there is a peak appeared at the same retention time for both of the sample solution and standard working solution. the GC/MSD confirmation test should be conducted.
46、The analyte could be confirmed when the relative abundance of its total monitor ions accord with that of the pesticide 11 SN/T 1591-2005 standards ion and the similaritdegree of their relative abundance ratio in土10%.Under the above GC/MSD operating conditions, the retention time of atrazine、vinclozo
47、lin、triflumizole、imazalil、buprofezin、propiconazole，fenarimol、pyridabenand the abundance ratio of the monitoring ions (m/z) of 9 heterocclic pesticides see to table 3. Table 3 Retention time and the abundance ratio of the monitoring ions of 9 heterocclic pesticides pestcides Retention time/min the ab
48、undance ratio of the monitoring ions atrazine 18.41 173 : 187 : 200 : 215(26 : 3 : 100 : 58) vinclozolin 21.09 187 : 198 : 212 : 285(74 : 89 : 100 : 86) procymidone 25.46 96 : 255 : 283 : 285 ( 100 : 8 : 69 : 5) triflumizole 25.67 219: 248 : 278 : 287(18 : 7 : 100: 53) imazalil 27.33 173 : 215 : 240
49、 : 296(76 : 100 : 9 : 6) buprofezin 28.30 105: 172 : 175: 305(100 : 35 : 25 : 6) propiconazole 32.20,32.58 173 : 191 : 259 : 261 (100 : 27 : 88 : 57) fenarimol 39.12 139 : 219 : 251 : 330(100 : 63 : 54 : 36) pyridaben 41.64 117 : 147: 309 : 364(15 : 100: 7 : 6) 3. 4. 4 81ank test The operation of the blank test is the same as that described in the method of determination. but with omission of sample addition. 3. 5 Calculation and expression of resul
