1、中华人民共和国出入境检验检疫行业标准SN/T 1967-2007 进出口食品中异稻瘟净残留量的检测方法Determination of iprobenfos residues in food for import and export 2007-08-06发布2008-03-01实施飞-P帝国椭-EA口中华人民共和国发布国家质量监督检验检疫总局SN/T 1967-2007 前言本标准的附录A、附录B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s中华人民共和国湖南出人统检验检疫局、中华人民共和向内蒙古出入境检验检疫局、中华人民共和国黑龙江出入境检验检疫局、中华人民
2、共和国福建出入境检验检疫局。本标准主要起草人z李拥军、颜鸿飞、张莹、黄志强、王美玲、李刚、杨长志、杨芳。本标准系首次发布的出人攻检验检疫行业标准。SNjT 1967-2007 进出口食品中异稻瘟净残留量的检测方法第一法气相色i昔质谱法1 范围本标准规定了食品中异稻瘟净的气相包i吉质i吉检测方法。本标准适用于茶叶、菠菜、莽头、苹果、板栗、蜂蜜、食醋、大米、鸡肉、牛肉、鱼肉中异稻瘟净残留量的测定和确证。2 方法提要试样中残留的异稻瘟净采用丙闹和正己Jt(l十2)振荡提取，石墨化碳黑固相萃取柱或中性氧化铝周相萃取柱净化，洗脱液浓缩并定容后，供气相色i吉质谱仪扭定和确证，外标法定量.3 试剂和材料除另
3、有规定外，所有试剂均为分析纯，水为蒸馆水.3. 1 正己烧z重蒸俑，3.2 丙囚E重蒸馆。3.3 氯化纳。3. 4 元水硫酸纳2经650C灼烧4h，置干燥器中备用。3.5 丙酬+正己统0+2)溶液。3.6 丙if.l+正己烧o十1)溶液。3.7 异稻瘟净标准物质(probenfos，CH 2I O，PS，CAS:26087-47-8):纯度大于99%。3.8 标准储备溶液g准确称取适量的异稻瘟净标准物质，用丙酣将配制成1000g/mL标准储备液，再根据检测要求用正己统稀释成相应的标准工作溶液。标准溶液避光于4C保存。3.9 石墨化碳黑固相萃取柱:3mL, 125 mg，或相当者。3. 10 中
4、性氧化铝固相萃取柱:3mL.125 mg.或相当者。4 仪器和设备4. 1 气相色谱-质i苦联用仪，配电子轰击离子源(EI源), 4.2 组织捣碎机。4.3 粉碎机。4.4 涡旋混匀器。4.5 固相萃取装置，带真空泵。4.6 多功能微量化样品处理仪，或相当者。4.7 低速离心机:3000 r/min, 4.8 离心管:15mL。4.9 刻度试管:15 mLo 4. 10 微量注射器:10L。1 SN/T 1967-2007 5 试样制备与保存5. 1 试样制备5. 1. 1 水果或蔬菜取有代表性祥品500g.将其可食用部分切碎后不可用水洗涤，用捣碎机将样品加工成浆状。1昆匀，装入洁净的盛样容器
5、内，密封并标明标记.5. 1.2 茶叶及粮谷取有代表性样品500g，用粉碎机粉碎并通过2.0mm困孔筛.混匀，装入洁净的盛样容器内，密封并标明标记.3mL丙副十正己烧混合液(3.6)洗脱萃取柱，保持流速1.5 mL/min.收集全部流出液，于45C下，氮气流吹至近于。最后用正己:tJt定容至0.5mL.供GC-MS分析。6.3 测定6.3. 1 气相色i昔-质谱条件2 a) 包i苦柱，HP-5MS石英毛细管柱.30mXO. 25 mm(内径).膜厚0.25m.或相当者;b) 色谱柱温度2初始温度80L:.以7L:/min升至205C.再以25C/min升至280L保持5min; d 进样口温度
6、，280L;d) 包i苦质i昔接口温度，270L;e) 载气:氮气，纯度大于等于99.995%.0. 8 mL/min; f) 进样量，JL;SN/T 1967-2007 g) 迸样方式2元分流进样，1min后开阀ph) 电离方式:EI;i) 电离能量:70eV; j) 检测方式z选择离子监测方式CSIM); k) 监测离子Crn/z):203.204.246.288;定量离子:204;) 溶剂延迟:10min ., 6.3.2 色谙测定与确证根据样液中异稻瘟净的含最情况，选定降面积相近的标l1t工作溶液，对标准工作液和样液等体积参插进样，测定标准工作溶液和样液中异稻瘟净的响应值均应在仪器检测
7、的线性范围内。在相同实验条件下，样品中待测物质的质量包谱保留时间与标准工作液相同，并且在扣除背景后的样品质量色讼中，所选离子均出现，经过对比所选择离子的丰皮七与标准品对应离子的丰度比，其值在允许范白内(允许范用见表1)贝jiT判主样飞I.f在J泣的f，n!jl功J在第6.3.1条规定的包i昔条件下，异稻瘟净的参考保留时间是17.hb:，;tr;，;主w.d117-(m/z)丰度比单203:204 246 288 17: 100 15 : 19。色说图和邱吉图参见Fdfkii 时相对离毫丰度最大容许误差相对丰度(基峰)/%GC/MS时相对离子丰度最大允许误差/%20-50 10-20 10 土1
8、5:t20 土506.4 空白实验除不加试样外，均按上述测) 1 ( . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . JJFJY J 7. 1 测定低限本方法的测定低限为0.005mg/kg. 7.2 回收率添加浓度和回收率见表2。表2回收率数据样品名称添加浓度/(mg/kg)回收率范围/%0.005 91. 9-113.1 大米0.01 91. 6-1饵.1 0.20 97.5-119.。3 SN/T 1967-2007 表2(续样品名称添加改度l(mg/kg)回收率范围1%0.005 74.9-93.3 茶
9、叶0.01 90.8-107.9 0.02 83.5-108.1 0.005 86.8-110.1 菠菜0.01 91. D-110. 5 0.02 107.1-118.9 0.005 75.3-86.5 苹果0.01 92.8-119.9 0.02 95.6-113.1 0.005 96.8-114.4 蜂蜜0.01 97.7-115.8 0.02 91. 2-112. 8 0.005 109.9-120.9 板栗0.01 90. 2-108. 8 0.02 88.8-119.5 0.005 93.4-104.2 食醋0.01 89.9-100.6 0.02 95.2-107.2 0.005
10、 87.6-98.5 奔头0.01 93.7-100.6 0.02 82.1-9.1.3 o. (旧581. 8-10.1. 9 鱼肉0.01 80.9-94.0 0.02 90.0-105.8 0.005 86.4-凹.4牛肉0.01 92.4-11 1. 0 0.02 91. 2-11 1. 9 0.005 70.9-103.0 鸡肉0.01 70.2-93.1 0.02 73.3-92.7 第二法气相色谱法/ B 范围本标准规定了食品中异韬瘟净残留莹的气相色1苦测定方法。本标准适用于大米、菠菜、苹果、牛肉、鸡肉、鱼肉、蜂蜜、板栗、茶叶、食即等食品中异稻瘟净残留flf4 SN/T 1967
11、-2007 的测定。, 9 原理、r试样中残留的异稻瘟净农药经闪酬+正己炕飞1+2)和正己烧提取，过固相萃取柱净化，用配备火焰光度检测器的气丰1色谱仪进行测定，外标法定量。10 试剂与材料除另有规定外，所有岚剂均为分析纯，水为蒸馆水。10. 1 正己炕重蒸倍。10.2 丙削重蒸惰。10.3 元水硫酸的:经650C灼烧4h，置干燥器中备用。10.4 丙酬+正己炕(+2)1在液。10.5 异稻瘟净标准物质(probenfos，CH O，PS.CAS，26087-47-8)，纯度大于99%。10.6 标准储备济液z准确称取适茸的异衍瘟净标准物质，用丙酬将配制成1000月/mL标准储备液，再根据检测要
12、求川正己炕稀将成相应的标准工作溶液。标准溶液避光于4C保存。10.7 石墨化碳照固相萃取扛一:3mL , 125 rng，或相当者。10.8 中位氧化知固相萃取柱，3mL. 125 mg.戎相当;哀。11 仪器和设备11. 1 气相色谱仪2配火焰光!立检J!mCFPD。11. 2 快速混匀器。11. 3 离心机，3000 r/min. 11. 4 多功能做最化样品处理仪，jj)tfU当者。11. 5 具东刻度离心管，5mL，10mL。11. 6 玻硝试管，20mL. 11. 7 尖畹吸管。11. 8 微量可调移液器，10L.200L.1000L. 11. 9 微量注射器，10L。12 试样制备
13、与保存12. 1 试样制备12. 1. 1 水果或蔬菜取有代表性样品500g，将其可食用部分切碎后不可用水洗涤，ffj捣碎机将样品加工成浆状。j昆匀，装人iti净的应样容器内.密封并标明标记。12. 1. 2 茶叶及粮谷取有代表性样品500g，用粉碎机粉碎并通过2.0mm圆孔筛。混匀，装入i古净的盛样容器内，密封并标明标记。12. 1. 3 肉及肉制品取代表性样品500g，将其切碎后，用捣碎机将样品加工成浆状，混匀，装入ti净的盛样容器内，密封:Jio标明标记。12.2 试样保存茶叶、蜂产品、调味品及粮谷类等试样于0C4C保存:水果蔬菜类和肉及肉制品类等试样于一18C以下冷冻保存。5 SN/T
14、 1967-2007 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。电F13 测定步骤13. 1 提取与净化13. 1. 1 对于大米、板栗、蜂蜜样品，称取2g(精确至0.001g)均匀试佯于10mL离心试管巾，加入2 mL水，加入元水硫股纳使之饱和，加入2mL丙副+正己炕混合液00.4)振荡提取两次，每次2min , 然后离心3min(离心速度为2000r/min).吸取上层提取液于刻度离心;残渣再用2mL正己炕提取两次，合并提取液于刻度离心管中，在多功能微量化样品处理仪或其他相当的仪器上，于40C下用氮气流吹至1.0 mL.供进样分析。13. 1. 2 对于菠菜、苹果、食酣
15、样品，称取2g(精确至0.001g)均匀试样于10mL离心试管中。对于茶e) o g) h) 尾吹气，20mL/min; 。进样为式元分流进样.1.0 min后开阀，j) 进样最，2L。13.2.2 色谱测定根据样液中异稻应净含量情况，选定与样j在浓度相近的标准工作溶液。异稻瘟净标准工作溶液和样液中异稻瘟11.响应倪均应在仪器捡到HJt性范围内。标准工作溶液和样液等体积穿插进样测定，以保留时间定性，测量峰面积与标准工作液比较进行定量。在上述色i吉条件下，异稻瘟净标准物质的色谱图参见附录B中图B.1.13.3 空白试验除不加试样外，按上述测定步骤进行。6 SN/T 1967-2007 13.4
16、结果计算和表述用色谱数据处理机或按式(2)计算试样中异稻瘟净残留景，计算结果需扣除空白值。式中=X=AXcxV = As X m X一样品中异稻瘟净含量，毫克每千克(mg/kg); A一一样液中异稻瘟净的峰面识;As 标准工作溶液中异稻瘟净的降面积;c 标准工作溶液中异稻瘟净浓度，微克每毫升g/mU;V 样液最终定容体积，毫升(mL); m一一称取的试样质量.克(g) 14测定f:限回收率r 1 14. 1 B月旧定低限I I ( 2 ) 本方法在大米、菠菜、苹果、牛肉、鸡肉、鱼肉、驿蛮、板栗和食醋E测定低限为0.005mg/kg;在茶叶r 表3回踱军费据样品名称添加w度I(mg/kg)回收率
17、范固1%J齿。.0.0且. 75.0-94.0 大米0YM古气81. 0-106 / f. 2 飞88.0-92.5 /J .005 92.2-102 4 菠菜0.01 / 、82.3-108 / 川2呼/ 82.0-98.5 .10叫/ 74.8-91. 2 苹果o.oJ ! 78.5-98.1 1. / 80.5-105 50 20-50 Permitted tolerances!% 士10:t 15 10-20 士2010 士506. 4 81ank test The operation of the blank test is the same as that described in
18、 the method of determination. but with omission of sample addition. 6. 5 Calculation and expression of result The calculation of iprobenfos content in the sample is carried out by GC!MS data processor or ac cording to the following formula (1The blank value should be subtracted from the above result
19、 of calculation x= A Cs V Asm ) 1 ( . . . . . . . . . . . . . . . . . . . . . . where X-Iprobenfos content in the sample.mg!kg, A-Peak area of iprobenfos in the sample solution; cs-Peak area of iprobenfos in the standard working solution，g!mL; As一Concentrationof iprobenfos in the standard working so
20、lution; V一Finalvolume of sample solution. mL; m-Mass of test sample.g. 7 Limit of determination and recovery 7.1 Limit of determination The limit of determination of this method is 0.005 mg!kg. 16 S:I/T 1967-2007 7. 2 Recovery According to the experimental data, the fortifying concentration of isopr
21、ofos for each sample and its corresponding recoveries see table 2. Table 2-The recovery of the method Sample Spiked concentration/ (mg/kg) Range of recove叩1%0.005 91.9-113.1 rlce 0.01 91.6-108.1 0.20 97.5-119.0 0.005 74.9-93.3 tea 口.019日.8-107.90.02 83.5-108.1 0.005 86.8-110.1 leek 0.01 91.9-110.5 0
22、.02 107.1-118.9 0.005 75.3-日6.5apple 0.01 92.8-119.9 0.02 95.6-113.1 0.005 96.8-114.4 hone 0.01 97.7-115.8 。.0291.2-112.8 0.005 109.9-120.9 chestnut 。0190.2-108.8 0.02 88.8-119.5 0.005 93.4-104.2 vlnegar 0.01 89.9-100.6 0.02 95.2-107.2 0.005 87.6-98.5 allium chinense 0.01 93.7-10日60.02 82.1-94.3 0.0
23、05 81.8-104.9 fish 0.01 80.9-94.0 0.02 90.0-106.8 17 SN/T 1967-2007 Table 2 (continued) Sample Spikedncentration/ (n咱IkgJRange of recovery/ % 0.005 86.4-99.4 beef 0.01 92.4-111.0 0.02 94.2-111.9 0.005 70.9-103.0 chicken 0.01 70.2-93.1 0.02 73.3-92.7 The secon4111ethod Gas chromatography method 8Scop
24、e/j / / and confirFatlon。fiprobenfos resldes by gas chromatog-raphy-mass spect rometry in food.(;r- This standard is applicable for derr_inat旧roflsoprof口sresidue inl巾e，spmac 9 Principle 10 Reagents and materials ，-_.，. acet9协hexane(1 + 2) and hexane. ter / / Unless otherwise specified.all reagents u
25、sed should be of analytical grade. water is distilled water or corresponding de-ionized water. 10. 1 Acetone. 10.2 n-Hexane. 10.3 Anhydrous sodium sulfate, Ignite at 650(; for 4 h.and keep in a desiccator. 10.4 Acetone-n-hexane solution,Acetone+ n-hexane (1 + 2). 18 SN/T 1967-2007 10.5 1probenfos st
26、andard (C比1O,PS.CAS,26087.47.8) , Purity99%. 10.6 Iprobenfos standard solution, Accurately weight an adequate amount of iprobenfos standard. dissolve in a small volume of acetone. Dilute with acetone to form a standard stock solution of 1 000g/mL in concerntration. Then dilute the standard stock sol
27、ution with n-hexane to the re quired concerntration as the standard working solution. The standard solution should be stored below 4C and keep in dark place 10.7 Graphitized carbon black cartridge,3 mL.125 mg.or equivalent. 川N阳阳阳川川e创削圳叫叫u川山川叫tr阳时r阳a剖|川 11川Aq刑刷u川叫i怡帆p川叫m吨电j / / 11. 1 Gas chromato口rap
28、h(equi口口edwithetector FPD)/ 11.2 Vortex mixer. / 11.3 Centrifuge,3 000 r/min 11.4 Multifunction sample treatment unit for micrpch回响tca.1method or equivalent. /、., / :二二工:二:1:;:;:止rr:丁:ffff怕川川h忖忖g伊竹r11.7 p酬illa川叫川W刑ry-tii11.8 11.9 12 Preparation and storage of test sample 12. 1 Preparation of test sa
29、mples 12.1.1 Fruits and vegetables The representative samples is reduced to ca 500日.which has been removed shell. seed. peel. stem. root.coronal (do not wash by water). then cut up the edible port旧nsare blended and then homoge nized thoroughly in a high speed blender .and then are placed in a clean
30、container as the test sample. 19 SN/T 1967-2007 which is sealed and labeled 12.1.2 Tea and grains or cereals The representative samples is reduced to ca 500 g. which is crushed with a grinder and let wholly pass through 2. 0 mm sieve , and then are placed in a clean container as the test samp怡，which
31、 is sealed and labeled. 12. 1. 3 Meats and meat products The representative samples is reduced to ca 500 g. The eatable potions are thoroughly ground and ho mogenized in a meat grinder. And then are placed in a clean container as the test sample , which is sealed and labeled. 12. 2 Storage of test s
32、amples The test samples of tea , bee products , grains Or cereals should be stored below ot -4t. The test samples of fresh fruits,vegetables.meat and meat products should be stored below -18C. In the course of sampling and sample preparation. precaution must be taken to avoid contaminat旧nor any fact
33、ors which may臼usethe change of residue content. 13 Procedure 13.1 Extraction and cleanup 13.1.1 For rice.chestnut.bee honey.weigh 2 9 of the test sample (accurate to 0.001 g) in 10 mL centrifuge tube. Add 2 mL water, then saturate the sample with anhydrous sodium sulfate. Add 2 mL acetone + hexane (
34、10.4) , blend for 2 min in vortex mixer and centrifuge for 3 min. Repeat the proce dure with 2 mL acetone + hexane (10. 4) twice, Then , the residue are extracted with 2 mL hexane , blend for 2 min in vortex mix肘，and centrifuge for 3 min. Repeat the procedure with 2 mL hexane twice.Combine the upper
35、 layer to obtain the extracted solution. Transfer all the extracts to centri fuge tube with ground stopper and concentrate to 1. 0 mL with nitrogen at 40t. The concentrated solution is ready for gas chromatographic determination. While emulsified during extracting. increase the extract solution volu
36、me 13.1.2 For Spinach.apple.vinegar.weigh 2 9 of the test sample (accurate to O. 001 g) in 10 mL centrifuge tube. For tea , weigh O. 5 9 of the test sample (accurate to O. 001 g) in 10 mL centrifuge tube ,add 2 mL water,then saturate the sample with anhydrous sodium sulfate. Add 2 mL acetone+ hexane
37、 (10.4) , blend for 2 min in vortex mixer and centrifuge for 3 min. Repeat the procedure with 2 mL acetone+ hexane (10.4) twice, Then , the residue are extracted with 2 mL hexane. blend for 2 min in vortex mixer,and centrifuge for 3 min. Repeat the procedure with 2 mL hexane twice.Com bine the upper
38、 layer to obtain the extracted solution. Elute the graphitized carbon black cartridge with 20 SN/T 1967-2007 4 ml acetone+ hexane (10.4). Discarded the eluent. Transfer all extracts to the graphitized carbon black cartridge.then elute the cartridge with 4 ml acetone+ hexane (10.4). Collect all the e
39、luent in centrifuge tube with ground stopper and concentrate to 1. 0 ml with nitrogen at 401巳.The concen trated solution is ready for gas chromatography determination 13.1.3 For beef.chicken.fish.weigh 2 9 of the test sample (accurate to O. 001日)in 10 ml centri fuge tube. Add 2 ml water. then satura
40、te the sample with anhydrous sodium sulfate. Add 6 ml of ac etone + hexane (10.4) blend for 2 min in vortex mixer and centrifuge for 3 min. Then. the residue are extracted with 2 ml hexane. blend for 2 min in vortex mixer. and centrifuge for 3 min. Repeat the procedure with 2 ml hexane twice. Combin
41、e the upper layer to obtain the extracted solution. Elute the neutral aluminum oxide cartridge with 4 ml acetone + hexane (10.4). Discarded the eluent. Transfer all extracts to the neutral aluminum oxide cartridge. then elute the cartridge with 4 ml ace tone + hexane (10.4). Collect all the eluent i
42、n centrifuge tube with ground stopper and concentrate to 1.0 ml with nitrogen at 40l:. The concentrated solution is ready for gas chromatography determina tion 13.2 Determination 13.2. 1 GC operating conditions a) Column , EQUITyTM_1701 used quartz capillary column.30 m x O. 32 mm x 1.0严m(i. d) .or
43、equiv alent , b) Column temperature ,lnitial temperature 100l: .ramp at 10l:/min to 220C .hold for 10 min, c) Injection port temperature,250C , d) Detector temperature,250l:, e) Nitrogen，Purity99. 99% .carrier gas 5.0 ml/min, f) Hydrogen,75 ml/min, g) Oxygen , 100 ml/min, h) Make-up gas,20 ml/min, j
44、) Injection mode , splitless purge after 1 min, j) Injection volume，2L 21 SN/T 1967-2007 13. 2. 2 GC determination According to the approximate concentration of isoprofos pesticide in the test sample solution. select the standard working solution with similar peak area to that of sample solution. Th
45、e responses of iso profos pesticide in the standard working solution and sample solution should be within the linear range of the instrumental detection. The standard working solution should be injected randomly in be tw臼nthe injections of sample solution of equal volume. Identify by the RT and quan
46、tify with peak area. Under the above GC opening condition. the Gas Chromatogram of isoprofos standard is seen fig ure 8. 1 in Annex B. 13. 3 81ank test The operation of the blank tes with omission of sample additi tlon. 14 Limit of determination and recovery 14.1 Limit of determi阳tion( 2 ) The limit
47、 of determination in rice. spinach. apple. cattle muscle. chicken musc怡.fish musucle. bee honey.chestnut and vinegar by this method is 0.005 mg/kg;tea is 0.01 mg/kg. 14.2 Recovery According to the experimental data. the fortifying concentration of isoprofos for each sample and its 22 SN/T 1967-2007
48、corresponding recoveries see table 3: Table 3-The revery of the method Sampl. Spiked concentration/ (mg/kg) Range of the revery/% 0.005 75.0-94.0 nce 0.01 81.0-106 0.2 88.0-92.5 0.005 92.2-102 leek 0.01 82.3-108 0.02 82.0-98.5 Z-0P4.-句0Q0但T5马-品油罚一卡也jY呻-74.8-91.2 apple 78.5-98.1 /且02.7 80.5-105 / O.吵
49、5/93.8-106 beef / /501 86.5-104 :2. 山古苦苦87.5-112 0.0051二.84.0-101 chicken 0.01. 85.0-102 / 产业三三毛x81.0-103 / I 0叫:7194.6-100 fish .O俨飞二二?86.0-117 口.中叮飞M 臼. 86.0-108 飞/ O. 05 / 1. 俨唁. 号h飞旷， 82.6-91.6 w Bee honey ， 86.6-105 片。2/ 77.0-92.0 /0.005 回-伊/81.8-104.9 chestnut 0.01 80.9-94.0 0.02 90.0-106.8 。.005
