1、 中华人民共和国出入境检验检疫行业标准SN/T 1971-2007 进出口食品中苟虫威残留量的检测方法气相色谱法和液相色谱质谱/质谱法Determination of indoxacarb residues in food for import and export-GC and LC-MS/MS methods 2007-08-06发布2008-03-01实施注一阳VF咽叫嗣中华人民共和国发布国家质量监督检验检疫总局SN/T 1971-2007 前-一一同本标准的附录A、附录B、附录C均为资料性附录。本标准由国家认证认可监督管理委员会提出并归口。本标准起草单位中华人民共和国福建出人境检验检疫
2、局、中国检验检疫科学研究院、中华人民共和国江苏出入境检验检疫局、食品安全分析与检测技术教育部重点实验室(福州大学)。本标准主要起草人z杨方、卢声字、徐敦明、李淑娟、沈72饪、余孔捷、林永辉、文IJ正才、陈国雨。本标准系首次发布的出人统检验检疫行业标准。1 范围进出口食品中部虫威残留量的检现u方法气相色谱法和液中目色谱-质i昔/质谱法第一法气相色谱法本标准规定了食品中苟虫威残留最检测的制样和气相色i由检测方法。SN/T 1971-2007 本标111适用于毛豆、青梗菜、袖子、生姜、木耳、笋、茶叶、猪肉、鱼肉、鸡肉中部虫威残rfi量的测定。2 方法提要试丰手中残留的ti虫威采用丙阳正己烧混合溶剂振
3、荡提取.ENVI-CARB回扣1萃取柱净化植物源性产品、Floriil回相萃取柱净化动物源性产品，1Jt带有电子捕获检测器的气相色谱仪测定，外标法定量。3 试剂和材料除非另有规定.所用试开IJ均为分析纯，水为主蒸水。3. 1 正己炕z色i昔纯。3. 2 丙剧:色谱纯。3. 3 乙肪=色谱纯。3. 4 氯化的。3.5 中性轼化铝z层忻用.100日200日。3. 6 饱和氯化纳溶液z称取过量氧化纳溶于水中。3. 7 丙酬+正己炕(+2)说介溶剂:盘取100mL r久i附和200mL正己:院，混匀。3.8 苟且L威标准物质:lndoxacarb.分子式C日CIF，N，O，.CAS 174060-41
4、-4.分子量527.剖，纯度大于等于98%。3.9 苟虫成标准储备液(100mg/U:准确称取迈量珩虫威标准物质，用丙酬溶解并定容至100mL.该标准储备液i可于4C冰箱中保存。3. 10 苟且成标准工作液:根据需要取适量标准贮备液，以正己炕稀释成适当浓度的标准工作液。标准工作液需现配现用。3. 11 ENVI-Carb 1M相萃取柱:250mg , 3 mL，用前以3mL丙自TJ+正己悦。十2)活化，保持柱体112润。3. 12 Florisil 目相萃取柱:500mg.3 mL. 4 仪器和设备4. 1 气相色谱仪，自己也子捕获检测器.4.2 离心机:4000r/min. 4.3 粉碎机。
5、4.4 组织捣碎机。4.5 涡旋混合器。4.6 超声波沂洗器。4. 7 元泊真空茶。SN/T 1971-2007 4.8 罔相萃取装置。4.9 氮吹仪。5 试样制备和保存5. 1 试样制备5. 1. 1 水果蔬菜类取有代表性样品500g.将可食部分切碎后(不可水洗).用组织捣碎机将样品加工成浆状.昆匀，装人沽11容器内，密封并标识。5. 1. 2 茶叶、食用菌(干)类取有代表性样品300g.用粉碎机粉碎并通过2.0mm困孔筛川昆匀，装人洁净容器内，密封并标识。5. 1. 3 肉及肉制品Z 取有代表性样品500g.切碎后丹缸织捣碎机将样品加工成浆状，混址，装入洁净容器内，密封并标识。6. 1.
6、1 茶叶、食用菌(干的样品z称.1.00g均匀试样.置于50mL具EE塑料离心管中，加入2mL饱和氮化纳溶液。.6).力1人3时，丙自三十iE己院混合溶剂。.7).涡徙振荡m混匀30s.超声提取20min , 以2500内m离心3min，取上层有#布宇穷可1fr:-交部市到1tl3时，丙用十正己炕混合时IJ(3. 7)重复提取两次，合并提取液，于35C7ls，W氮吹浓坊约1m-r 6. 1.2蔬菜与水果样品s称取10.00画眉匀t样，主ff50 ml具塞曲闲心管中，加5g ;化纳，再加6.2. 1 对于按6.1.1与6.1.2步骤提取的军tfz将样品提取E豆豆ENVI-CARBJl;l相萃取柱
7、，保持流速约为1mL/min.当样液液面接近固相萃取柱J料层时，用6mL丙丽十正己;院混合溶剂10-20 +-30 表3布虫威药物残留的添加回收率数据样曰:名称t在力1浓度/(mgkg)0.005 青梗菜0.010 0.020 。005生莹0.010 0.020 10 土50回收率/%I03.0117.6 91. 4-117. 4 96. 8 108.5 H9. 2-117. 6 94.8-118.1 93.3-103.2 7 SN/T 1971-2007 表3C续)样品名称添加放度/(mg/kg)回收率/%。.00577.8-98.6 茶叶0.010 82.3-2.3 0.020 82.9-
8、95.0 0.005 80.2-112.6 笋0.010 108.3-118.9 0.020 93.9-105.8 0.005 105.0-9.8 柏于0.010 86.0-3.8 0.020 90.7-100.7 0.005 86.0-113.6 毛豆0.010 103.0-9.2 0.020 98.1-8.0 0.005 67.4-93.8 木耳0.010 72.3-2.3 0.020 72.5-92.1 0.005 87.4-100.2 猪肉o. ,-qO 82.3-2.3 。.u20 93.7-106.5 。.00586.6-5.8 鸡肉0.010 71. 2-102. 9 。.020
9、86.3-100.4 0.005 66.2-88.4 鱼肉0.010 94.1-104.1 0.020 85.9-102. 1 8 SN!T 1971-2007 附录A(资料性附录)苟虫威的气相色谱图ECD1 A. 293. 1 7. 640.:( 2.50 3.CO 3.5。如004.50 5.00 MRM of 3 Channels ES+ 528.2249.1 9. 66e4 00 4.50 5.00 图c.11部虫戚的多反应监测(MRMl离司色i曹图11 SN/T 1971一2007Foreword Annex A.Annex B.Annex C of this standard ar
10、e informative annexes This standard is proposed by and is under the jurisdiction of the Certification and Accreditation ad ministration of the Peoples Republic of China_ This standard is drafted by Fujian Entry-Exit Inspection and Ouarantine Bureau of the People 5 Re public of China. Chinese Academy
11、 of Inspection and Ouarantine. Jiangsu Entry田ExitInspection and Ouarantine Bureau of the People s Republic of China. Key Laboratory of Analysis & Detection Tech nology for Food Safety (Fuzhou Universityl Ministry of Education. Main drafters of this standard are Yang Fang. Lu Shenyu. Xu Dunmin. Li Sh
12、ujuan. Sheng Chongyu. Yu Kongjie. Lin Yonghui. Lin Zhengcai.Chen Guonan. This standard is a professional standard promulgated for the first time. Note: This English version is a translation from the Chinese text.is solely for guidance 12 SN/T 1971-2007 Determination of indoxacarb residues in food fo
13、r import and export -GC and LC-MS/MS methods The first method Gas chromatography method 1 Scope The standard specifies the method of determination of indoxacarb residue in foodstuffs for import and export by gas chromatography. The method is applicable to determine indoxacarb residues in green soybe
14、ans. qing.gen-cai. grape fruit .ginger. fungi. bamboo shoot. tea. pork. fish and chicken for import and export. 2 Principle Extraction of the indoxacarb residues with acetone and n-hexane mixed solvents. and then purifica tion with a ENVI-CARB solid phase extraction (SPE) cartridge for plant origin
15、food.Florsil solid phase extraction (SPE) cartridge for animal origin food. followed by gas chromatography analysis 3 Reagents and Materials Unless otherwise specified .all regents used are A. R. .and pure water is redistilled water. 3. 1 n-Hexane: HPLC grade. 3.2 Acetone: HPLC grade. 3.3 Acetonitri
16、le: HPLC grade. 3.4 Sodium chloride. 3.5 Neutral aluminum oxide: 100 mesh-200 mesh. 3.6 Saturated sodium chloride solution: Add sodium chloride in water to the solution is saturated. 3.7 Acetone and n-hexane mixed solvent:Pipet 100 mL actone and 200 mL n.hexane and mix well. 13 S:I/T 1971-2007 3.8 S
17、tandard of indoxacarb, Molecular formula C H17 CIF, N,o, ,CAS 174060-41-4, molecular weight 527. 84，purity二三四0/0.3.9 Stock solutions of indoxacarb (100 mg/L) ,Accurately weigh indoxacarb standard material ,dis-solve with methanol to a volume of 100 mL,and store at approximately 40C for a maximum per
18、iod. 3.10 Calibration solutions of indoxacarb for GC , Dilute appropr旧tevolume of stock solutions to a intended concentrat旧nwith n-hexane and mixed 队lell.These solutions should be prepared before use. 3.11 extraction cartridge was condi-revent the columns from runing dry. 3.12 Florisil solid phase e
19、xtracti 4 Apparatus and equipment 4. 1 4.2 Centrifuge,4 000 r/min. 4.3 Grinder. 4.4 Tissues homogenizer. 4.5 Vortex mixer. 4.6 Ultrasonic machine. 4. 7 Vacuum pu付1口.4.8 Solid phase extraction equipment 4.9 Pressured gas blowing concentrator. 14 S:-;jT 1971-2007 5 Sample preparation and storage 5. 1
20、Sample preparation 5. 1. 1 Furits and vegetables Collect ca 500 9 the representative samples the edible portions are cut up (without washing by wa terl and mixed well with a tissure homogeniz町.then sealed in clean containers and marked. 5.1.2 Tea and fungi Collect ca 300 9 the representaive samples
21、and crush with a 1)叫er，let them pass through 2. 0 mm sieve , and then sealed in clean Collect ca 500 9 the representat mogenizer, and then sealed in cIan c门tilinersand morked. 5.2 Sample storage 6 Method of Determinatibn 6. 1 Extraction 6. 1. 1 For tea and fungi: Weigh 1. 00 9 of the prepared test s
22、amples into a 50 mL stoppered plastic centrifge tube , then add 2 mL saturated sodium chloride solution (3.6) and 3 mL acetone and n.hex ane mixed solvent (3. 7) , stopper the tubes and votrtex lor 30 s and extract with ultersonic machine for 20 min , and then centrifuge at 2 500 r.min lor 3 min. tr
23、ansler the supernatant to another clean tube and repeat the extraction procedure again. The supernatants are collected in a 3 mL tube and evaporat to ca 11L at 350C under a stream of nitrogen. 6.1.2 For vegetables and lurits:weigh 10.00 9 01 the prepared test samples into a 50 mL sto口peredplastic ce
24、ntriluge tube.add ca 5 9 sodium chloride and 10 mL acetone and n-hexane mixed solvent (3. 7) ,extract with ultersonic machine lor 20 min ,and then centriluge at 2500 r/min lor 3 min ,trans ler the supernatant to another clean tube and repeat the extraction procedure agai门Thesupernatants are collecte
25、d in a 10 mL tube and evaporat to ca 1 mL at 350C under a stream 01 nitrogen. SN/T 1971-2007 6.1.3 For meat and meat products:weigh 10.00 9 of the prepared test samples into a 50 mL stop pered plastic centrifuge tube,add ca 5 9 sodium chloride and 10mL acetone and n-hexane mixed sol vent (3.7) ,extr
26、act with ultersonic machine for 20 min,and then centrifuge at 2 500 r/min for 3 min. transfer the supernatant to another clean tube and re口eatthe extraction procedure again. The super natants are collected in a 10 mL tube and evaporated to ca 1 mL at 35 C under a stream of nitrogen. and re-extract t
27、he analyte with 3 x 2 mL acetonitrile,collect the acetonitrile phase and evaporat to ca 1 mL at 35 C under a stream of nitrogen 6. 2 Clean-up 6.2.1 For samples extracted according to 6. 1. 1 and 6. 1.2: Pass extracts through ENVI-CARB car tridges conditioned at a flow rate ca 1 mL/min, discard elute
28、s , then the cartridges are dried under vacuum ,and the indoxacarb residues are eluted with 6 mL acetone and n.hexane mixed solvent (3. 7). The elutes are collected and evaporated to dryness at 350C under a stream of nitrogen and re dissolved in 1 mL acetone for GC-ECD analysis. 6.2.2 For samples ex
29、tracted according to 6. 1.3: Connect Florisil PR cartridges to a solid phase ex traction equipment,fill 0.5 9 neutral aluminum oxide in the top of the cartridegs,and condition the SPE cartridegs with 3 x 1 mL acetone and n-hexane mixed solvent (3.7) at a flow rate ca 1 mL/min. Pass extracts through
30、Florisil PR cartridges and elute with 6 mL acetone and n-hexane mixed sol vent. The elutes are collected and evaporated to dryness at 35C under a stream of nitrogen and re dissolved in 1 mL acetone for GC-ECD analysis. 6.3 Determination 6. 3. 1 GC operation conditions a) Column:HP-5 fused quartz cap
31、illary column,30 m x O. 32 mm(i. d.) ,film thickness 0.25m or equivalent columns, b) Column temperature:lnitial temperature 80C , ramp at 20 C /min to 180C , hold for 3 min , ramp at 20 C /min to 240C ,hold for 3 min ,and then ramp at 20 C /min to 270 C ,hold for 13 min, c) Injection port temperatur
32、e:220C , d) Temperature of detector:280C , e) carriergas:Helium,purity99.995% ,flow rate is 1 mL/min, f) Injection volume:2L， g) Injection mOde:Splitless , purge after 1 min 6.3.2 GC/ECD detection Prepare standard solutions containing indoxacarb at appropriate concentrations according to the ana Iyt
33、e in sample extracts. The standard solutions should be injected in between the injections of sample 16 SNjT 1971-2007 solution of equal volume. The responses of standard solutions and sample solutions should be within the linear range. The referenced retent旧ntime of indoxacarb is 24. 8 min. Annex A
34、is the chromato gram of the standard solution. 6.4 Calculation and expression of the result Calculate the concentration of the indoxacarb residues in sample according to the formula (1) , vx-m !/T 1971-2007 Annex A ( Informative) Gas chromatogram of indoxacarb standard solution fEECE-EDIA. (20061lFY
35、C飞.I1l02.D) IIz 700 hdy-TNA川川尸ili-600 500 25 mm 27 26 Figure A. l-Gas chromatogram of indoxacarb standard solution 25 24 T 23 22 T 21 T 20 S:TjT 1971-2007 Reference mass conditions, a) Capillary,O.5 kV , b) Source Temperature, 120C, Annex B ( Informative) Reference mass conditions c) Desolvation Tem
36、perature，4骂。C，可d) Cone Gas Flow (Nitrogen) ,5 L/h . e) f) Collision Pressure (Argon) ,2, g) Precursor Compound (m/z) indoxacarb 528.2 Note: The daughter ion with .脊be different, and the MS par L品Cone/V 30 30 CE/eV 13 16 1 ) Noncommerc旧Istatement:the reference mass parameters in Annex A are accomplis
37、hed by waters UPLC/Priemer LC.MS/MS, the equipment and i!s type involved in the standard rnethod is only for reference and not related to an commercial aim.and the analysts are encouraged to use equipments of different corporation or different type 26 Annex C (Informative) SN(T 1971-2007 Multiple reaction monitoring chromatogram of indoxacarb standard solution 中华人民共和因出人境检验检疫行业标ll迸出口食品中部虫威残留量的检测方法气相色谱法和液相色i昔-质i昔/质谱法SN/T 1971-2007 * 中罔标m出版社出版北京复兴门外三里河北衔16号邮政编码;1000-15 网址电:68S2394668517548 中国标准出版社秦皇岛印刷厂印刷争等开本880X 1230 1/6 印张2字数19干字2007年11月第版2007斗11月第一次印刷印敖1-2000 书号155066.2-18252 定价15.00元
