1、中华人民共和国出入境检验检疫行业标准SN/T 1986-2007 进出口食品中滇虫睛残留量检测方法2007-08-06发布J毡雪公Determination of chlorfenapyr residues in food for import and export 2008-03-01实施整二平中华人民共和国发布V酬可国家质量监督检验检菇总局中华人民共和国出入境检验检疫行业标准迸出口食品中漠虫睛残留量检测方法SN!T 1986-20口7 中国标准出版社出版北京复兴门外三里向北街16号邮政编码，100045网址电话，6852391668517548 中国标准出版社秦皇岛印刷厂印刷句h开本880
2、X12301/16 印张1.25 字数30千字20口7年11月第一版2007年11月第次印刷印数1-2000句陡书号:155066.2-18271 定价12.元SN/T 1986-2007 目。白本标准的附录A为资料性附录。本标准由国家认证认可监督管理委员会提出并归口。本标准起草单位2中华人民共和国天津出入境检验检疫局、中华人民共和国浙江出人挠检验检疫局、中华人民共和国福建出入挠检验检疫局、中国检验检疫科学院。本标准主要起草人:林安清、张豆、许可4、吴延晖、章鄂、何佳、古珑、肖亚兵、鲍晓霞、杨方、李淑娟.本标准系首次发布的检验检疫行业标准。I SN/T 1986-2007 进出口食品中i臭虫睛
3、残留量检测方法1 范围本标准规定了食品中澳虫脂残留量检测的制样，气相色谱检测方法和气相包谱-质谱确证方法。本标准适用于红豆、茶叶、毛豆、甘蓝、苹果、醋、鸡肉、猪肉、鱼肉、牛肉等澳虫腊残留量测定。2 方法提要试样中残留的澳虫精采用乙脂提取，中性氧化铝固相萃取柱或石墨化炭黑固相萃取柱净化，洗脱液浓缩后定容，供气相色谱测定和气相俨质诸仪确证，外标法定量。3 试剂和材料除另有规定外，所用试剂均为分3. 1 乙腊z高效液相色谱级。3.2 正己烧z高效液相色谱级。3. 3 丙阴z高效液相色谱级。3.4 元水硫酸销2经650C灼烧4h, 3.5 氯化纳。3.6 3.7 注.标准储备液和标准工作液在0飞;P4
4、C叶林自中保匠。3. 12 中性氧化铝固相萃取柱，3ml,5oo th)o, 3. 13 石墨化炭黑固相萃取柱，3mL500 mg , 3. 14 元水硫酸纳柱，80mmX40 mn议内径筒形漏斗底部酸纳。飞血气, _.;.7 4 仪器与设备4. 1 气相色谱仪，带电子捕获检测器。4.2 气相色i苦-质谱仪，带负离子化学源。4.3 拙滤装置。4.4 旋转蒸发器。4.5 氮吹仪。4.6 高速均质器。4. 7 离心机3000 r/min, 4.8 团相萃取装置。4.9 ，民旋混合器。SN/T 1986-2007 5 样品和J备与保存5. 1 样品制备5. 1. 1 水果及蔬菜类取苹果、毛豆、甘蓝等
5、有代表性样品约500g，将其可食用部分切碎后，用捣碎机将样品加工成浆状。i昆匀，装人洁净的容器内，密闭并标明标记。5. 1. 2 茶叶及粮谷类取茶叶、红豆、粮谷等有代表性样品约500g，用粉碎机粉碎并通过孔径2.0mm圆孔筛。j昆匀，装入洁净的容器内，密闭并标明标记。5. 1.3 动物源食品将猪肉、鸡肉、牛肉、水产品等有代表性样品约500g用绞肉机绞碎，昆匀，装人洁净的容器内，密闭并标明标记。5.2 试样保存茶叶、调味品及粮谷类等试佯于4C以下保存;水果蔬菜类和动物源食品等试样于一18C以下冷冻保存。在制样过程中，应防止样品受到污染或发生澳虫脂残留量的变化。6 测定步骤6. 1 提取6. 1.
6、 1 动物源样品称取5g(精确到0.01g)试样于50mL离心管巾，加10g无水硫酸纳(3.4).20 mL提取液(3.8), 均质2min, 3 000 r/min离心5min。将溶液转移至250mL分液漏斗中，重复上述操作一次。合并提取液于上述分液漏斗中，静置10mino将乙脑层分出并通过无水硫酸纳柱(3.14)至250mL浓缩瓶中，再分别将20mL和10mL乙脂加入分液漏斗中，振摇5min，静置10mn.分层。合并乙脐层于七述浓缩瓶中.40C水浴旋转蒸发至干。用5mL正己统济解残渣。6. 1. 2 植物源样品一般植物样品称取20日(精确到0.01g)，粮谷等含水分较少的植物样品称取10g
7、(精确到O.Olg)，茶叶称取5g(精确到0.01g)于150mL烧杯中，加入50mL乙脂，均质2min，经布氏漏斗抽滤至加有10 g氯化纳(3.5)和20mL磷酸盐缓冲溶液(3.7)的250mL具塞锥形瓶中。以50mL乙脂洗涤烧杯和均质器，合并乙脂洗液于布氏漏斗中并抽滤收集于上述锥形瓶中，猛烈振摇5min，静宜15mi口。将乙脂层定容至100mL，取50mL乙腊层至250mL浓缩瓶中，在40C水浴旋转蒸发至干。在浓缩瓶中加入O.5时，氯化纳溶液(3.6) , 20 mL正己坊，超声1min.振摇2min，将样液通过无水硫酸纳柱(3.14)至另一150mL浓缩瓶中，用20mL和10mL正己烧洗
8、涤浓缩瓶两次，过上述无水硫酸纳柱，40C水浴中旋转蒸发至约5mL, 6.2 净化6.2. 1 动物源样品将6.1. 1中得到的正己烧样液全部通过中性氧化铝国相萃取柱(3.12，使用前依次用3mL丙阳和3 mL正己;院预洗)，以5mL正己统洗浓缩瓶，一并过柱。再用3mL洗脱液(3.9)洗脱，收集全部洗脱液于收集管中，在40C下用氮吹仪浓缩至干。用1.0 mL正己烧溶解残渣，供气相色谱仪测定和气相色谱-质谱确证。6.2.2 植物源样晶将石墨化炭黑固相萃取柱(3.13)和中性氧化铝固相萃取柱(3.12，使用前依次用3mL丙酬和3mL 正己炕预洗)自上而下安装，将6.12中得到的正己统样液全部通过固相
9、萃取柱，用5mL正己烧洗浓缩瓶，一并过柱，再用5mL正己烧淋洗固相萃取柱。弃去石墨化炭黑团相萃取柱，用3mL洗脱液(3.9)SN/T 1986-2007 洗脱中性氧化铝固相萃取柱，收集全部洗脱液于收集管巾，在40C下用氮吹仪浓缩至干.粮谷样品用1. 0 mL正己炕溶解残渣，茶叶样品用0.50mL正己烧溶解残渣，一般植物样品用2.0mL正己炕溶解残渍，供气相色谱仪测定和气相色i苦质i昔确证。6.3 测定6.3. 1 气相色i昔条件a) 色i古拉，DB5-MS搀性石英毛细管柱30mXO. 32 mm内径).膜厚0.25m.或相当者:b) 柱沮程序起始温度90C(保持1rnin) ，以10C/min
10、升温至270C(保持5min); c) 进样口温度:250C;d) 电子俘获检测器温度:300C;的我气z氮气，纯度大于等于99.999%.流速1.5 mL/min; ) 进样方式无分流，进样后0.7min打开分流阀;g) 迸样量1L。6.3.2 气相色谱-质谱条件a) 离子源z负离子化学源，甲炕反应气，b) t1:口沮度:250C ; c) 离子源温度:230C;d) 载气z氮气，纯度大于等于99.999%.流速1.0 mL/min; e) 进样方式2元分流，进样后0.7min打开分流i词;) 采集方式=全扫描:g) 进样量:2L;h) 选择离子(m/z):79.234.269.349, 6
11、.3.3 气相色i曹测定根据样液中澳虫肪含最情况，选定峰顶积相近的标准工作溶液。标准工作溶液和样液中澳虫肪响应值均应在仪器检测线性范围内。标准工作溶液和样液等体积参插进样测定。在上述色谱条件下，澳虫腊的保留时间约为17.0min.标准品的色i昔因参见附录A中图A.l。6.3.4 气相色谱质谱法确证标准济液及样液均按6.3.3规定的条件进行测定，如果样液中与标准溶液相同的保留时间有峰出现，则对其边行质谱确证。经确证分析被扭tl物质1I也谱峰保留时间与标准样品相一致，并旦在扣除背景后的样品i古医l中，所选择的离子均出现g同时所选择离子的丰度比与标准样品相关离子的相对丰度一致.相似度在允差之内(见表
12、).则可判定样品中存在澳虫师。被确证的样品可判定为j臭虫脂阳性检出。标准品的总离子流图参见附录A中图A.2.标准品的质i古图参见附录A中图A.3.表1定性确证时相对离子丰度的最大允许偏差相对离子丰度1%允许的相对偏差1%50 2050 1020 10 士20: 25 :!: 30 +50 6.3.5 空白试验除不加试样外，均按上述步骤进行。6.3.6 结果计算和表述用色i出数据处理机或按式()计算试样中汲虫脂残留量，计算结果应将空白值扣除。) 1 ( . . . . . . . . . . . . . . . . . . . . . . . . . 式中2X 试样中澳虫腑的残留壶，单位为毫克每
13、千克(mg/kg); 3 SN/T 1986-2007 A 样液中澳虫脆的峰面积pAs 标准工作溶液中澳虫腊的峰面积;Cs一一-标准工作溶液中澳虫腊的浓度，单位为微克每毫升仰自ImL)，V 样液员终定容体积，单位为毫升(mL)，m 最终样液所代表的试样量，单位为克(g)o7 方法的测定低限、回收率7. 1 测定低限气相色谱法和气相色谐-质谱法的测定低限均为0.005mg/kg。7.2 回收率见表2。表2样品的添加阳回收吟验数据样品猪肉鸡肉鱼肉牛肉茶叶4 添加浓度!(问!kg主阳川!样品拉加浓度!(问/以5 10 20 5 10 20 5 10 20 5 10 20 5 10 20 72土工艺J
14、|气二r5 J骂:/黯lt(旷;矿f衍叭川，.!3川-旷 . 11 . ., 11 !引!:111;4;f;丁工工:Up亨严1有毛玄艾卢产口jtk飞j肌8I 11 ./ J:/ 去记咐.jV气，一/fL/ 5 10 20 5 10 20 5 10 20 回收率市围f%88.3-105.8 74.3-100.0 83.2-97.3 84.2-110.。96.8-102.7 92.3-107.4 77.9-90.8 72.9-91. 7 70.5-86.2 81. 5-94. 8 78.1-92.4 82.7-96.3 89.0-108.0 79.0-91. 6 81. 9-109. 0 SN/T
15、 1986-2007 附录(资料性附录)滇虫踊标准晶色谱图A ha罔画。-au-d问。协同ECDl A，归CJ1016旧500002.0)H, 4 500 4 000 mm 22.5 20 17.5 / ff?!图3 500 3 000 2 500 二一2 000 1 500 1 000 NL: 1. 056 TICMS xcjl00ng 5 2. 5 500 。/ r飞/ / 喝蝇归卢./23.58 19.8820.2520.4721. 52 22.49 17.2811 82 8 6 8 4 5 4 3 3 3 回12 21 2 。2 RT:ll. 48-23. 98 100 95 90 8
16、0 75 70 85 65 60 45 50 55 UUF85A主王们wdE40 35 30 25 20 15 10 5 。5 23 22 21 20 澳虫腊标准品总离子流图(NCI)19 17 18 Timc (min) 16 15 图A.214 13 12 SN/T 1986-2007 fish-50-1 #802 RT: 16. 85 AV:l NL:L 21E5 T: O. Ol-c Cldet500. 00 Fu!l ms 55. 00-475. 00 100 95 90 85 80 75 70 65 吕60;55 2目击450丘40100 150 232.4 200 348.9 3
17、47.0 350.9 269.1 312.0 352. 1 250 300 350 m , 图A.3漠虫信标准品质谙图CNCI)6 396.2 句伊.，.400 435.7 460.8 吁甲.，.450 SN/T 1986-2007 Foreword Annex A of this standard is an informative annex. This standard was proposed band is under the charge of China National Regulatory Commission for certification and Accreditat
18、ion. This standard was drafted by Tianjin Entry-Exit inspection and Ouarantine Bureau of the Peop怡sRepublic of China. zhejiang Entry-Exit inspection and Ouarantine Bureau , fujian Entry-Exit inspection and Ouarantine Bureau.Chinese academy of inspection and quarantine. The main drafters of this stan
19、dard are Lin Anqing. Zhang Man , Xu Hong. Wu Yanhui , Zhang Hua. He Jia. Gu Long.Xiao Yabing.Bao Xiaox旧.YangFang. Li Shujuan. This standard is a professional standard promulgated for the first time. 7 SN!T 1986-2007 Determination of chlorfenapr residues in food for import and export 1 Scope This sta
20、ndard specifies the methods of sample preparation and determination by GC and GC.MS of Chlorfenapr residue in foods TI川叫r时d叫p州Ii阳阳蛐c臼创a甜b恤l怡e叫le阳阳t怡err盯巾Ybean. cabbage. apple. vinega，-chicken俨pork号L鄂shand beef. ) 2 Principle 3 Reagents and materials / l Un巾less。州t仙阳he圳川阴s邻阳机口阳阳e町cifie旧眈eWater. f 1 ,
21、 3. 1 Acetonitrile, HPLC grae. 3.2 n.Hexane, HPLC grade. 3.3 Acetone, HPLC grade. ,;r d乡，;J Y j/ , /i / 3.4 Anhydrous sodium sulfate, ignite at 650t for 4 h. and store in air.tight container. 3.5 Sodium chloride. 3.6 Sodium chloride solution (100 g/U , weigh 100 9 of sodium chloride. add water to ma
22、ke a 1 L solution. 3.7 Phosphate buffer (1 mol/L pH 7.0) , Dissolve 105 9 (土O.1日)of dipotassium hydrogenphos. phate (几HPO，)and 61 9 (土0.1g) of potassium dihydrogenphosphate (KH, PO,) in approximately 8 SN/T 1986-2007 500 mL of water. Adjust the pH of the solution to 7. 0 with 1 mol/L sodium hydroxid
23、e or 1 mol/L hydrogen chloride. Add water to make a 1 L solution. 3.8 Extract solvent: .n-Hexane saturated by acetonitrile- acetonitrile (1 + 1, V / V). 3.9 Elute solvent: n-Hexane - acetone (1 + 1 , v / V) 3.10 Chlorfenapyr standard (C15Hll SrCI凡几0，CAS: 122453-73-0): Purity99%. 3. 11 Chlorfenapyr s
24、tandard solution: Weigh appropriate chlorfenapyr. dissolve in n-Hexane and 4 Apparatus and equipm,eft I 4.1 GC: Equipped with E10, / 4.2 GC-MS: Equipped witt NCI. 4.3 Suction filter system. 4.4 Rotation evaporator. 4.5 Nitrogen evaporator with heated bath. 4.6 Homogenizer. 4.7 Vortex mixer:3 000 r/m
25、in 4.8 Solid phase extraction equiment. 4.9 Minishaker. ) funnel , filled with 20 9 anhydrous / 9 SN/T 1986-2007 5 Sample preparation and storage 5. 1 Sample preparation 5. 1. 1 Fruit and vegetable Reduce the fruit or vegetable sample of apple.green soy bean and cabbage to ca 500 9 by quartering. mi
26、x thoroughly. place in clean containers. seal and label. 5. 1.2 Tea and cereal Reduce the tea or cereal sample to ca 500 9 by quartering. grind thoroughly and let pass through a 20-mesh sieve.mix thoroughly.place in clean containers. seal and label. 5. 1.3 Animal original food Reduce the animal orig
27、inal food sample 01 pork. chiken. beel and aquatic口roductto ca 500 9 by quar tering. mix thoroughly. place in clean containers. seal and label 5. 2 Storage 01 test sample The test sample 01 tea. condiment and cereal shall be stored below 4C; fruit. vegetable and animal original lood shall be stored
28、below -1St. In the course of sample preparation. precaution should be taken to aviod contamination or any factors which may cause the change 01 residue content. 6 Procedure of determination 6. 1 Extraction 6. 1. 1 Animal original food Weigh 5 9 (accurated to O. 01 g) of the test sample into 50 mL ce
29、ntrifuge tube. add 10 9 of anhy drous sodium sulfate (3.4) and 20 mL extract solvent (3.8) .homogenize 2 min. centriluge 5 min at 3000 r/min. Taking the organic phase into 250 mL separator lunnel , repeated above operation Combine the organic phase. wait 10 min. Collect the acetonitrile phase and pa
30、ss through the cloumn of anhydrous sodium sullate (3.14) to 250 mL erlenmeyer flake. Rinse the separator lunnel with 20 mL and 10 mL acetonitrile .shake 5 min. let stand 10 min lor separately comperately. Combine ace tonitrile phase.rotary evaporate the extraction liquid at 40t to dry. dissolve the
31、residues with 5 mL n-Hexane. 10 SN/T 1986-2007 6. 1.2 Plant original sample Weigh 20 9 (accurated to O. 01日)of the almost plant test sample .or 10 9 (accurated to O. 01 g) cereal. or 5 9 (accurated to 0.01 g) tea into 150 mL beaker .add 50 mL acetonitrile.homogenize 2 min. filter with Suction filter
32、 system funnel to 250 mL taper bottle with 10 9 sodium chloride (3.5) and 20 mL phosphate buffer (3.7). Rinse the baker and homogenizer and filter to the same taper bottle. Shake 5 min.wait 15 min. Distilled the acetonitrile phase to 100 mL. Pipet 50 mL to 250 mL erlenmeyer flake. rotary evaporate a
33、t 40 C to dry. Add O. 5 mL sodium chloride solution (3.6) and 20 mL n-hexane. ultrasonic 1 min.shake 2 min.pass this liquid through the column of anhydrous so dium sulfate (3. 14) to another 150 mL erlenmeyer flal哩.rinse the erlenmeyer flake twice with 20 mL and 10 mL n-hexane.pass through the same
34、column of anhydrous sodium sulfate. rotary evaporate at 40 C to about 5 mL 6.2 Clean up 6.2. 1 Animal original food Transfer above solution (6.1.1) to the alumina oxide neutral cartridge (3.12. preconditioned with 3 mL acetone followed by 3 mL n-hexane). Rinse the erlenmeyer flake with 5 mL n-hexane
35、 .pouring the solution into the cartridge. Then elute the column with 3 mL elute solvent (3.9) .and collect all the elution in a tube and evaporate to dryness under nitrogen flow at 40t. Make up to 1.0 mL with n-hexane for GC or GC-MS determination. 6.2.2 Plant original sample Set up the graphitized
36、 carbon column (3.13) and the alumina oxide neutral cartridge (3.12. precon ditioned with 3 mL acetone followed by 3mL n-hexane) from up to down . Transfer above solution (6.1.2) to the cartridge. Raise the erlenmeyer flake with 5 mL n-hexane pouring the solution into the cartridge. Wash the cartrid
37、ge with 5 mL n-hexane . Discard the graphitized carbon column. Then elute the alumina oxide neutral column with 3 mL elute solvent(3. 9) .and collet all the elution in a tube and evaporate to dryness under nitrogen flow at. 40C. Dissolve the residues with for GC or GC-MS determination. With 2.0 mL n
38、-hexane for almost plant test sample. with 1.0 mL n-hexane for cereal. and with O. 50 mL n-hexane for tea 6.3 Determination 6.3. 1 GC operating conditions a) Column , DB5-MS 30 m x O. 32 mm( i. d. ) filn、thickness0.25m Or equivalent, b) Column temperature,90 C for 1 min.ramp at 10C/min to 270 C .hol
39、d for 5 min, c) Injection port temperature,250 C , 11 SN/T 1986-2007 table 1). We can confirm that there are corresponding analyte in the sample. See figure A. 2 and figure A. 3 in annex A Table l-Maximum permitted tolerance for relative ion intensities using a range of mass spectrometric techniques
40、 Relative intensity (base peak) / % GC-MS (relative) :!: 25 10-20 :!:30 10 :!: 50 50 土2020-50 6.3. 5 Blank test ) 1 ( . . . . . . . . . . . . . . . P g/mL. 7 Limit of determination and 7. 1 Limit of determination The limit of determination of this method of GC and GC- MS is O. 005 rng/kg. 7. 2 Recov
41、ery See table 2. 13 SN/T 1986-2007 Table 2一Thefortifyingncentration of chlorfenapyr in samples and its corresponding recoveries fortify cncentration reCQve叩10同ifcncentrationreCQvery sample g/kg)C%) 回mple(glkg) (% ) 5 72.4-102.8 5 88.3-105.8 阳rk10 84.1-107.4 10 74.3-1.0 20 92.0-11 1. 2 20 83.2-97.3 5
42、 80.5-84.5 5 84.2-110.0 chicken 10 86.2-97.4 四bb句e10 96.8-102.7 20 81.5-101.5 20 92.3-107.4 5 79.5-98.0 5 77.9-90.8 fish 10 73.0-90.1 red bean 10 72.9-91.7 20 74.4-93.3 20 70.5-86.2 5 90.2-109.6 5 81.5-94.8 beel 10 83.2-106.8 apple 10 78.1-92.4 20 90.5-100.5 20 82.7-96.3 5 94.2-109.8 .5 89.0-108.0 t
43、ea 10 88.2-105.8 green soy 10 79.0-91. 6 20 88.5-103.5 20 81. 9-109.0 14 SN/T 1986-2007 Annex A (informative annex) Chromatogram of standard EEEJ恒。-FQ=。=ECDl A，【XCJlOI队DB500002D)H, 4 500 4 000 3 500 3 000 2 500 2 000 1 500 1 000 500 ffim 22守S20 17.5 Figure A. l-Gas chromatogram of chlorfena口yrstanda
44、rd 5 12.5 0 7.5 5 2.5 。NL 1. 05E6 TICMS xcjl00ng 16.85 RT: l l. 48-23. 98 00 95 90 85 80 75 70 65 60 out-mRUE2PZE55 50 45 40 8 1 6 1 4 8 4 1 1 5 4 1 3 3 3 1 3 8 12 21 2 1 1 0 2 1 35 30 25 23.58 19.8820,_ 25 20.4721. 52 22.49 17.2817.82 20 5 0 5 。23 22 2 Figure A.2-GC.MS (NCI) of TIC chlorfenapyr sta
45、ndard 20 9 17 18 Time (minl 6 5 4 3 2 SN/T 1986-2007 fish-50-1 #802 RT: 16. 85 AV:l NL: 1. 27E5 丁:0，O-c Cldel=500. 00 Full ms 55. 00-475. 001 100 95 90 85 RO 75 70 65 0 i80 255 350 , 击45u 40 t 7 ;111爪1l丁rL|f1L91川叫0100 150 269.1 200 250 300 miz 348.9 347.0 350.9 312.0 352.1 396.2 .俨矗，.-400 435.7 460.8 叩.，.450 350 Figure A. 3-Mass spectrogram of chlorfenapyr standard (NCI) 川一元9-ut一2嗣-2 m E1 6-6-nRU-5-lE ZES 号一价书一定
