1、E 中华人民共和国出入境检验检疫行业标准SN/T 1981-2007 进出口食品中环氟茵肢残留量的检测方法气相色谱-质谱法Determination of cyflufenamid residues in food for import and export GC-MS method 2007-08-06发布2008-03-01实施且也-w中华人民共和国发布国家质量监督检验检疫总问中华人民共和国出入境检验检疫行业标准迸出口食品中环氟茵肢残留量的检测方法气相色i昔-质谱法SlT 19812007 替中国标准出版社出版北京军兴门外三里河北街16号邮政编码，!OOO5网址.Cn 电话:6852394
2、668517548 中国标准出版社秦皇岛印刷厂印刷晤开本880X12301116 印张1.25 字数33千字2007年11月第一眼2007年11月第次印刷印数12 000 书号;155066.2-18266 定价12.00元SN/T 1981-2007 前言本标准的附录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:中华人民共和国山东出入境检验检疫局、中华人民共和国浙江出人统检验检疫局。本标准主要起草人=李立、李征、尹大路、丁慧瑛、曹鹏、李秀勇、防涛。本标准系首次发布的出入境检验检疫行业标准。I 1 范围进出口食品中环氟茵肢残留量的检测方法气相色i昔-质谱法本标准
3、规定了食品中环氟菌胶残留量的制样和气相色谱质i古检测方法。SN/T 1981-2007 本标准适用于小麦、玉米、花生、波萝、苹果、青葱、胡萝札紫苏叶、金银花、姜粉、花椒粉及茶叶中环氟茵胶残臼量的检测和确证。2 方法提要样品用提取，提取制un入氯化帆水后.JHE川arh/LC叫#1必C18固相萃取柱或弗罗里硅土层析柱净化，气相色i肯质i古法测定-:-;环节路口i?/ 3 试剂和材料/ ,/ 除另有规定外，试剂均为色i昔纯，必挤占过N水。/3. 1 乙肺。3.2 正己:tJt。3.3 丙酬。3.4 甲苯。3.5 乙赂。3. 6 氮化俐z优级纯。3. 13 Envi-Carb/LC-N H，固相萃取
4、柱，500mg/500 mg;, 3.14 C8固相萃取柱，1000 mg , 3.15 弗罗里硅土柱z破硝层析柱30cmX 15 mm(内径).中下端依次装入1cm高的无水硫酸纳、10g 弗罗里硅土和1cm高的无水硫酸纳。4 仪器和设备4. 1 气相色说-质i吉联Jll仪配有电子轰击离子源(EJ源)。4.2 振荡器。4.3 抽走涡混合器。4.4 旋转蒸发仪。SN/T 1981-2007 4.5 高速均质器。4.6 离心机。4.7 离心管，100mL。4.8 分液漏斗，150mL. 4.9 滤膜，0.45m.5 试样的制备与保存5. 1 试样制备5. 1. 1 小麦、米、金银花、茶叶取有代表性
5、样品约500g.Jll粉碎机全部粉碎并通过2.0mm圆孔筛。混匀，装入洁净的容器内，密闭，标明标记.5. 1. 2 水果或蔬菜取有代表性样品约500g.将其可食用部分切碎后，用食品捣碎机将样品加工成浆状。1昆匀，装入洁净的容器内.密闭，标明标记。5. 1. 3 姜粉、花椒粉取有代表性样品约100g，充分混合均匀.过2.0mm因孔筛，装入洁净容m内，密闭，标明标记。5. 1.4 :花生取有代表性样品500g，用磨碎机全部磨碎。J昆匀，装入洁净的容器内，密闭，标明标记。5.2 试样保存小麦、玉米、花生、茶叶、姜粉、花椒粉试悖于OC-4(:保tf;水果和蔬菜类试样于18C以下玲冻保存。在拙样及制样的
6、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。6 测定步骤6. 1 提取6. 1. 1 粮谷、坚果类称取10g试样(精确至0.01g)于100mL离心管中，加20mL水，放置5mino加50mL乙腑，于10 000 r/min均质提取1min，离心3min，将提取液移人100mL容jl瓶中。残留物再分别用2X20mL 乙脂重复提取两次。合并提取液.用乙脐定容至100mL.准确吸取20mL提取液于150mL分液漏斗中，依次加入10g氯化的和20mL磷酸缓冲液，振摇3mn。静置后，弃去水层。6. 1. 2 水果、蔬菜f，j:取20g试样(精确至O.01 g)于100mL离心管中，加50mL
7、乙脯，于10000 r/ min均质1min, 离心3min，将提取液移入100mL容量瓶中，残留物再分别用2X20mL乙肪重复提取两次，合并提取液，用乙精定容至100mL。准确吸取20mL提取浓于150mL分液漏斗中，依次加入10g氯化伪和20 mL磷酸缓冲液，振jJi3 min.静置分层，后，弃去水层。乙脐层加入5g无水硫酸纳脱水后于450C旋转浓缩至近干，加入2mL乙睛十甲苯(3+)混合溶液溶解。6. 1. 3 调味料、中草药称取5g试样(精确至0.01g)于100mL离心管中，加10mL 水，放置15min。加50mL乙腑，于10000 r/ min均质1min，离心3min，提取液移
8、入100mL容量品中，残渣再分别用2X20mL乙脂重复提取两次，合并提取液，用乙n定容至100mL.准确移取20mL提取液于150mL分液漏斗中，依次加入10g氯化伪和20mL磷酸缓冲液，振摇3min。静置分层，弃去下层水液。乙肪层加l人5g元水硫酸的脱水，于45.C浓缩至近干.fIl正己炕定容至5mL。6. 1. 4 茶叶称取试样1g(精确至0.01g)于50mL离心管中，加人1g氯化纳和3时，水，在淤涡混合器上充2 SN/T 1981一2007分混匀1min，放置。.5h，加入10mL正己炕+丙自由0+1)混合溶液，以10000 r/ min均质1min，离心3min，吸取上层有机相过无水
9、硫自主例柱于浓缩瓶中，残渣再分别用2X20mL正己炕+丙酬。+1)重复提取两次，合并上层有机相，于45.C水浴中减压浓缩至近干。6.2 净化6.2. 1 粮谷、坚果用10mL乙脐预淋洗C18小柱，弃去流出液。注入乙脂提取液(6.1.1)，再用2mL乙脂洗脱，收集全部洗脱液，经元水硫酸纳脱水，于45.C浓缩至近干，用2mL乙腊十甲苯(3十)混合溶液溶解。用10mL乙脐+甲苯(3十1)混合溶液预淋洗Envi-Carl】/LC-NH，(500 mg/500 mg)小柱，弃去流出液。注人上述所得溶液，再用25mL乙肪+甲苯(3+1)混合溶液淋洗，收集洗脱液，于45(旋转浓缩至近干，残留物用1.0 mL
10、丙酬+正己统(1+1)混合溶液济解，过O.45m滤膜，供气相色i吉质谱测定。6.2.2 蔬菜、水果用10mL乙脐+币苯(3十1)混合溶液淋洗Envi-carb/LC-NH，(500 mg/500 mg)小柱，弃去流出液。注入6.l. 2所得提取液，再用25mL乙肪+甲苯(3+1)混合溶液淋洗，收集洗脱液，于45.C浓缩至近干，残留物用1.0 mL丙朋+正己统0+1)混合溶液溶解，过0.45m滤膜，供气相色i汗质谱测定。6.2.3 调昧料、中草药用30mL正己烧淋洗弗罗里硅土层析柱，弃去流出液。向柱中注入2mL 6. 1. 3的提取液，用100 mL正己烧+乙谜07十3)淋洗，收集洗脱液于45.
11、C以下旋转浓缩至近干，残留物用1mL丙用十正己统0+1)混合溶液溶解，过0.45m滤膜，供气相包谱-质i苦测定。6.2.4 茶叶用5mL正己炕溶解残渣，转移入弗罗里硅土柱中。用5mL正己烧淋洗，弃去流出液，用5mL正己炕+乙M(9+1)混合溶液洗脱。收集洗脱液于10mL离心管，在45.C水浴中用吹氮浓缩仪缓缓吹至近干，残留物用1mL丙网+正己炕(1+1)混和液溶解，过0.45m滤膜，供气相色谱质i古测定。6.3 测定6.3. 1 气相色谱-质谱条件a) 色i苦柱:DB-35MS石英毛细管柱:30mXO. 25 mm(内径)，0.25m(胶厚); b) 柱温:70.C旦旦旦.260.C0 min
12、)三卫士.300.C00 min); c) 近样口沮度:250.C;d) 辅助加热器:280C;e) 离子源混度:230.C ; f) 囚极杆温度:150.C ; g) 我气z氮气纯度二三99.999%, 1 mL/min; h) 迸样是:2L;i) 进样方式:脉冲不分流gj) 电离方式:EI;k) 电离能量:70eV; 1) 选择离子:(m/z)412、321、294、275，定量离子412。6.3.2 气相色i曹-质谱检测及确证根据样液中被测物含量情况，选定浓度相近的标准工作洛液，对标准工作溶液与样液等体积参插进样测定，标准工作熔液和待测样液中环氟商胶的响应值均应在仪器检测的线性范围内。如
13、果样液与标准工作溶液的选择离子包谱图巾，在相同保留时间有色谱崎出现，并且在扣除背景后的样品质量色话中，所选离子均出现，所选择离子的丰度比与标准品对应离子的丰度比，其值在允许范SN/T 1981-2007 围内(允许范围见表1)。在6.3.1条件下.环氟茵胶保臼时间是10.0min，其监测离子(m/z)为m/z412、321、294、275(其丰度比为100: 70 : 85 : 50)对其进行确证;根据定量离子m/z421对其进行外标法定量。在6.3.1条件下，环氟茵胶标准物创气相包i泞质i昔总离子流包i昔图和全扫描质i肯图参见附录Ao表1使用定性气相色谱-质谱时相对离子丰度最大容许误差相对离
14、于丰度!yo允许的相对偏爱1%7 结果计算与表述50 土1020-50 土15用色谱数据处理饥或按式(1)计算样品中环氟茵胶残留景。式中=x一试样中环氟茵胶残位且盟主主主干克(刊的g)tA 样品溶液中环氟茵胶的峰面积;/ / As 一环氟茵胶标准工作溶液的峰面极俨今/,/ V 样品溶液段终定容体积，单i与毫:ll( rnL) c 环氟茵胶标准工作液自主浓度单位为1攻克每毫升(g/ml-): m 最终样液代表的试得质量，单位为克以)0 B 测定低限和回收率8. 1 测定低限。.41;二3加浓度及回收毒品实验2V添见iJ坪i萃的问表加惊样品名称5导浓度l!g/kg): k四16f / 花椒 。缸l
15、飞, ,/ / : / / S ,_. 姜粉20 40 5.0 胡萝10 20 5.0 青葱10 20 4 10-20 土20回收率/%84.75-106.45 82.10-107.78 89.06-101. 72 84.65-100.90 82.08-110.55 83.10-103.51 90.80-107.80 90.33-95.90 94.06-108.44 90. 10-95.40 83.20-99.90 。1.94-99. 55 ,; 1 0 :l: 50 . ( 1 ) SN/T 1981-2007 表2(续)样品名称添加浓度/(阳/kg)回收率/%5.0 92.30-101.
16、30 苹果10 91. 43-116. 93 20 92.08-108.51 5.0 97.00-104.50 菠萝10 94.53-103.23 20 90.01-109.50 5.0 80.90- 92.70 玉米二俨引，啧斗二了牛、哼00 民争86.65-101. 30 79.80-95.78 才P飞r;-)-奇h一二J劳81. 20-11 1. 40 小麦10/飞机v 85.25-92.05 zk卢iZ83.48-96.53 75.00-100.70 花生与之三二:2lO0手IJJIJ 区90.80-109.00 86. 13-118. 15 10 民主二!83.30-107.00 一
17、一茶叶20 77.60-104.80 ff 40 /tJ汇，F、81. 50-102. 0。/;尤310 / r入81. 60-98. 50 金银花I飞、j咛吗20 f vf飞F 80.40-93.12 扩/40 84.46-94.42 84.35-101. 70 紫苏tJY?174/ / Iuj 82.85-103.45 by 84. 14 - 98. 88 啊晶同J J 5 SN/T 1981-2007 附录(资料性附录)环氟茵肢标准物质选择离子流图A TJC:IOPP 0 11. 15 ! 川! lJ 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00
18、13.00 14.0015.0016.0017.0018.0019.00 min Abundance 4 4001 4 2001 4 000 3 800 3 600 3 400 3 200 3 000 2 800 2 600 2 400 2 200 2 000 1 800 1 600 1 400 1 200 I 000 800 600 400 200 Time _ 412 341 355 367 393 ) 360 380 400 420 毛Z坏氟茵肢标准昂选择离子流图321 294 275 223 188 图A.1118 55 ehF1，hhvii?14 lil-llIll-ll000000
19、0000000 )町阳阳阳阳阳阳瞅瞅阳mm阳阳mmmmm毗mmmmmMmmmmm-m田m扭曲目拍自由Hmmm因回剧剧而阳臼m回归咄咄阻四岛囚HMSt1111111-A z m 环氟菌肢标准晶选择离子质i昔图图A.26 SN/T 1981-2007 Foreword Annex A of this standard is an informative annex. This standard was proposed byand is under the charge of the Certification and Accreditation Admin stration of the Peo
20、ples Republic of China. This standard was drafted by Shandong Inspection and Quarantine Bureau and Zhejiang Inspection and Quarantine Bureau of the Peoples Republic of China. This main drafters of this standard are: LiLi.LiZheng. Yin Da Lu.CaoPeng. LiXiuYon日.SuiTao.This standard is a professional st
21、andard promulgated for the first time. Note: This English version , a translation from the Chinese text, is 50lelfor gu idance. 7 SN/T 1981-2007 Determination of cyflufenamid residues in food for import and export-GC-MS method 1 Scope f cyflufenamid residue in wheat. ginger. the power of bunge prick
22、ly ash and tea. 2 Principle and confirmation is made by GC-MS. using external 3 Reagents and materials hromtography pure. Water is redis-ti lIed water. 3.1 Acetonitrile. 3.2 n-Hexane 3.3 Acetone 3.4 Toluene 3.5 Ethyl ether. 3.6 Sodium chloride. 3.7 Anhydrous sodium sulfate:Analytically pure.ignite f
23、or 4 h at 650C and keep in a tightly con tainer. 8 SN/T 1981-2007 3.8 Phosphate buffer solution (0.5 mol/L.pH=7. 0) : Dissolve 52.7 9 dibasic potassium phosphate (几HPO，)and 30.2 9 of monobasic potassium phosphate in ca 500mL water.adjust to pH 7.0 with 1 mol/L sodium hydroxide solution or 1 mol/L hy
24、drochloric acid solution dilute to 1 L with water. 3.9 Florisil:O. 150 mm-O. 250 mm 3.10 Cyflufenamid standard:CHF5N，O， .CAS NO: 180409-60-3.purity98%. 3. 11 Cyflufenamid standard stock sulution: Weigh an asequate amount of cyflufenamid standard. dissolve in acetone-n hexane t。preparea soluti口nof 1.
25、0 mg/mL as the standard stock solution. stored at ot -4 t. r 3.15 4.1 4.2 4.3 Mixer. 4.4 Rotary vacuum evaporator. 4.5 High speed homogenizer. 4.6 Centrifuge. 4. 7 Centrifuge tube: 100 mL. 4.8 Separatory funnel: 150 mL. 4.9 Filter:0.45m. / 、h吨喃自，jtJ9 SN/T 1981-2007 5 Preparation and storage of test
26、sample 5. 1 Preparation of test sample 5.1.1 Wheat.m刮目.honeysuckle. tea Take approximately 500 9 of representative sample. Grind with a grinder to pass through a 2. 0 mm round-hole sieve. Mix thoroughly and placed into a clean container, sealed and labelled. 5. 1.2 Fruits and vegetables Take approxi
27、mately 500 9 of representative sample. The edible part are blended in a high speed blend 肘，mixthoroughly and placed into a clean container, sealed and labeled. 5. 1.3 The power of ginger. the power of hunge prickly ash Take approximately 100 9 of representative samp怡，mixthoroughly pass through a 2.
28、0 mm round hole sieve. Placed into a clean container, sealed and labeled. 5. 1. 4 Peanut Take ap口roximately500 9 of representative sample. Grind and mix thorough忡.placed into a clean container, sealed and labeled. 5.2 Storage of the test sample Test sample of wheat.maize.peanut,tea ,the power of gin
29、ger,the power of bunge prickly ash should be stored at a range of OC -4C. The test sample of fruits and vegetables should be stored below -18C. During sampling and sample preparat旧n，precaution should be taken to avoid contamination or any factors which may cause the change of residue content 6 Proce
30、dure 6. 1 Extraction 6. 1. 1 Cereals and nuts Weigh 10 9 of the test sample(accurate to O. 01日)into a 100 mL cent门fugetube,add 20 mL water, let stand for 5 min.add 50 mL acetonitrile, Extract for 1 min in a speed of 10 000 r/min , certrifuge for 3 min. Transfer the extract into a 100 mL volumetric f
31、lask. The residue was extracted by 2 x 20 mL acetonitri怡，combinthe extract , dilute to 100 mL with acetonitrile. Exactly transfer 20 mL extract 10 SN/T 1981-2007 into a 150 mL separatory funnel . Add 10 9 sodium chloride and 20 mL phosphate buffer solution shake for 3 min . Place aside for separatio
32、n. Discard the water phase. 6. 1.2 fruit and vegetable Weigh 20 9 of the test sample (accurate to O. 01 g) into a 100 mL centrifuge tube.add 50 mL aceto nitrile.Extract for 1 min in a high spead of 10 000 r/mi门.certrifuge for 3 min. Transfer the extract in to a 100 mL centrifuge tube. Extract the re
33、sidue two times with 2 x 20 mL acetonitrile combin the ex tract.dilute to 100 mL with acetonitrile. Exactly transfer 20 mL extract into a 150mL separatory fun nel. Add 10 9 sodium chloride and 20 mL phosphate buffer solution. Shake for 3 min. place aside for separation . Discard the weater phase. Ad
34、d 5 9 anhydrous sodium sulfate into acetonitrile phase. Con dense acetontrile to nearly dryness by a rotary evaporator at 45C. Dissolve the residue with 2 mL acetontrile-toluene(3 + 1 ). 6. 1. 3 flavoring. herbal medicine Weigh 5 9 of the test sample (accurate to O. 01 g) into a 100 mL centrifuge tu
35、be.add 10 mL water. place aside for 15 min. Add 50 mL acetonitrile. Extract for 1 min in a high spead of 10000 r/min.cer trifuge for 3 min. Transfer the extract into a 100 mL volumetric flack. Extract the residue two times with 2 x 20 mL acetonitrile. combin the extract. Dilute to 100 mL with aceton
36、itrile. Exactly transfer 20 mL extract into a 150 mL separatory funnel. Add 10 9 sodium chloride and 20 mL phosphate buffer solution. Shake for 3 min place aside for separation . Discard the weater phase. Add 5 9 anhydrous so dium sulfate into acetonitrile phase. Condense acetontrile to nearly dryne
37、ss by a rotary evaporator at 45t. Make up exactly to a volume of 5 mL with n-hexane. 6.1.4 tea Weigh 1 9 of the test sample (accurate to O. 01 g) into a 50 mL centrifuge tube.add 1 9 sodium chlo ride and 3 mL water . Mix thoroughly for 1 min with a mixer. place aside for O. 5 h. Add 10 mL n-hex ane-
38、acetone( 1 + 1). Extract for 1 min in a high speed of 10 000 r/min. Certrifuge for 3 min. The upper layer passing through an anhydrous sodium sulfate column . Extract the residue for two times with 2 x 20 mL n-hexane -acetone (1 +门.combin the upper layer and evaporated to nearly dryness at 45C. 6.2
39、Cleaning up 6. 2. 1 Cereals. nuts Rinse the C18 column with 10 mL acetonitrile.discard effluent . Load the solution (6.1.1) to column. And elute the column with 2 mL acetonitrile. AII eluates dehydrated with anhydrous sodium sulfate and evaporated to dryness at 45C. Dissolve the residue with 2 mL ac
40、etonitrile.toluene. Rines a Envi.Carb/LC-NH, (500 mg/500 mg) with 10 mL acetonitrile-toluene; Load the above solution 11 SN/T 1981-2007 to column. Elute the column with 25 mL acetonitrile-toluene (3 +门.collect the eluates and evaporat ed to nearly dryness at 45 C . Dissolve the regidue with 1. 0 mL
41、acetone-n-hexane (1 + 1) passing through a 0.45m filter.the solution is ready for GC-MS determination and confirmation 6.2.2 Fruit and vegetable Rines a Envi-Carb/LC-NH, (500 mg/500 mg)with 10 mL acetonitrile-toluene. Dissard the effluent. Load the extract(6. 1.2) to column. Elute the column with 25
42、 mL acetonitrile-toluene (3 +门.collect the eluates and evaporated to nearly dryness at 45C. Dissolve the residue with 1.0 mL acetone-n-hexane (1 + 1). passing through a O. 45m filter. the solution is ready for GC-MS determination and confir mation 口3f川川f们彻la附m叫叫m叫川n吨川巾g忖仇hee町呐巾r巾阳baRir阳h怆M川e时刊f刊川l。叫
43、risi削l阻国s创sil川l川|川um川mnw川比怕川h3Om叫L川m町n罔1estCd th叫1column. Elu巾川t恒e阳|川川1wi川1咒旧Om毗，L，川川n仆川7卜-h阳e仪呵an(-e础阳he(吭町削(圳(忏川1行7忏+川巾川咀盯叫e创luated to nearly dryness at 45 C. Di罔SSOlv电川f啊可eresidue with 1 mL acetone-n-hexane (1 + 1门).p阳as白smgf -/ through a 0.45m fi Iter. the soluti甲!fis ready for GC.M&1川口士201口Rela
44、tive intensity(base peak)/% 士507 Calculation and expression of the result Calculate the content of fipronil residue in the test sample by GC-MS data processor or according to the followed formula. X=AXCXV As X m ) 1 ( . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 SN/T 1981-2
45、007 where X-the residue content of cyflufenamid in the test sample ,mg/kg; A一thepeak height of cyflufenamid in the sample solution; As-the peak height of cyflufenamid in the standard working solution; c一theconcentration of cyflufenamid in the standard working solution，问/mL;V-the final volume of the
46、sample solution,mL; m-the corresponding mass of the test sample representing the final sample solution , g. 8 Limit of determination and recovery 8. 1 Limit of determination The limit of determination of this method , wheat , maize, peanut , pineapple , apple , s臼lIion，carrot is 0.005 mg/kg. Perilla
47、 frutescens, honeysuck怡，the power of ginger, the power of bunge prickly ash and tea is 0.010 mg/kg 8. 2 Recovery The experimental data of the concentrations of cyflufenamid in the fortified sample and its corre sponding recoveries are seen in table 2. Table 2-The experimental data of the concentrati
48、ons of cyflufenamid in the fortified sa、pleand its corresponding recoveries Sample Added concentrat ions/ (g/kg) Recove叩range/%10 84.75-106.45 the阳werof bunge prickly ash 20 82.10-107.78 40 89.06-101.72 10 84.65-100.90 the power of ginger 20 82.08-110.55 40 83.10-103.51 14 SN/T 1981-2007 Table 2(nti
49、nued) Sample Added concentrations/(g/kg) Recove叩range/%5.0 90.80-107.80 carrot 10 90.33-95.90 20 94.06-108.44 5.0 90. 10-95.40 S臼Ilion10 83.20-99.90 20 91.94-99.55 5.0 92.30-101.30 apple 10 91.43-116.93 20 92.08-108.51 5.0 97.00-104.50 pine apple 10 94.53-103.23 20 90. 01-109. 50 5.0 80.90-92.70 盯1aJZe10 86.65-101.30 20 79.80-95.78 5.0 81. 20-111.40 wheat 10 85.25-
