1、中华人民共和国出入境检验检疫行业标准SN/T 1947-2007 进出口食品中氟草定残留量的检测方法气相色谱-质谱法Determination of f1 uroxypyr residues in foods for import and export GC-MS method 2007-08-06发布2008-03帽01实施户居笔-号耶?中华人民共和国发布/ 国家质量监督检验检菇总局SN/T 1947-2007 . 目U古同本标准的附录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中华人民共和国吉林出入境检验检疫局、中华人民共和国云南出入境检验检疫局
2、。本标准主要起草人:王明泰、牟峻、马晓刚、宋立国、周晓、李爱军、常东华、芦春梅。本标准系首次发布的出入境检验检疫行业标准。I 1 范围进出口食品中氟草定残留量的检测方法气相色i昔m质谱法本标准规定了食品中氟草定残留莹的气相色说-质谱检测方法。SN/T 1947-2007 本标准适用于柑楠、苹果、波菜、白菜、玉米、糙米、牛肝、鸡肾中氟萃定残留莹的测定和确证。2 方法提要试样用乙酸乙酣提取以卡胶胶色识阳拉净化，再盹经氛罗里硅土斗斗Floris川n巾isi剖叫i气相色t谱古-质i谱吉仪检测，外标f定量皂3 试剂和材料/ / / 除另有规定外，所用试剂均为分析t身份、为二次芳:留水。3. 1 甲醇(残
3、臼级)。3.2 环己烧(残留级)。3.3 乙酸乙酣(残留级人3.4 正己;皖(残留级)。3.5 浓硫股(优级)。3.6 3.7 3.8 4. 1 4.2 4.3 4.4 4.5 具塞锥型瓶，250mLo 4.6 浓缩瓶:50mL、250mLo 4. 7 离心机。5 试样制备与保存5. 1 试样制备5. 1. 1 柑楠、苹果、菠菜、白菜;Y / / 作溶液。该溶液在oOC_40C冰箱取代表性样品约500g.将其可食用部分(不可用水洗)切碎后，用捣碎机将样品加工成浆状，混匀，SN;T 1947-2007 装人洁净容器，密封，标明标记。5. 1. 2 玉米、糙米取代表性样品约500g，用粉碎机粉碎，
4、混匀，装入洁净容器，密封，标明标记。5. 1. 3 牛肝、鸡肾取代表性样品约1烛，取可食部分，经捣碎机充分捣碎均匀，装入洁净容器，密封，标明标记。5.2 试样保存粮谷类、水果蔬菜类于0C4 C保存g其他类试样于一ISC以下冷冻保存。在抽样及制样的操作过程中，应防止样品受到污染或发生残留物含量的变化。6 测定步骤6. 1 摄取称取20g试样(精确至0.1g)于250mL具塞锥形瓶中，加入50mL乙酸乙附和20g无水硫般的(玉米、糙米除外)，均质提取3min，将提取液抽tJ于250mL浓缩瓶中。残渣再用50rnL乙酸乙EY重复提取一次，合并滤液，于40C7Jcl谷中浓缩至近干。加人5mL环己烧+乙
5、股乙固自(1+1)溶解残渣，待净化。6.2 净化6.2. 1 凝胶色谱净化(GPC)6.2. 1. 1 凝胶色谱条件a) 凝胶净化柱，700mrnX25 rnm , Bio Beads S-X3，或相当者$b) 流动相=环己烧乙酸乙1i(1+l), c) 流速，5.0mL/min, d) 样品定量环，5.0rnL，e) 预淋洗体积，100mL, f) 洗脱体积，260rnL , g) 收集体积，70rnL 130 mL. 6.2. 1. 2 凝胶色谱净化步骤将5rnL待净化液按6.2. 1. 1规定的条件进行净化，合并馆分收集器中的收集液(70mL 130 mL)于250mL浓缩瓶中，于40C
6、水浴中浓缩至近干，加入2mL正己:院以溶解残渣，待净化。6.2.2 回相萃取净化(SPE)使用前用6mL正己烧预淋洗弗罗里硅土(FlorisiD因相萃取柱，将样液顷入柱中，然后开j10 mL正己统+乙酸乙酣(7+3)进行洗脱.收集全部洗脱液于50mL浓缩瓶中，于40C水浴中浓缩至近干。用2 mL甲醉溶解残渣并转移至50mL具比色ff巾，待Ii化。6.3 黯化6.3. 1 样品醋化加1.5 mL浓硫酸于50mL具塞比色管中，在95C水浴中酣化10min，取出冷却，加10mL硫酸纳水溶液，同时准确加入5mL正己饶，振荡2min，取上层正己炕提取液，离心3min，供气相色i苦质ir测定。6.3.2
7、标准溶波醋化取适量的标准工作溶液，加2mL甲踪后，按照6.3.1的步骤W化。6.4 测定6.4. 1 气相色谱-质谱条件a) 色i苦柱，HP-1701石英毛细管柱，30mXO. 25 mm(内径)X 0.25m，或相当者;20 C /mn Or.， ., 5C /min b) 包i贯注温度，50C(2min)一一一一180C(1 min)一一一270C(18 min) , 2 c) 进样口温度:280C;d) 色消-质i肯接口温度:280C;时我气:氮气，纯度大于等于99.999%，流速1.0 mL/min; f) 进样量:1L;g) 进样方式g无分流进样，1.5 min后打开分流阀;hl 也
8、离方式:EI;i) 电离能量:70eV; j) 测定方式:选择离子监测方式gkJ 选择监测离子(m/z):定最209，定性209、181、211、268;) 溶剂延迟:5，minc 6.4.2 气相色谙-质谱检测及确证SN/T 1947-2007 根据样液中氛革定含量情况，选定浓度相近的标准工作溶液，标准工作溶液和待测样液中氟草定的响应值均应在仪器检韧的线性范围内。标准工作溶液与样液等体积参捅进样测定。标准溶液及样液均按6.3.1规定的条付进行测定，如果样液中与标准溶液相同的保留时间有峰出现.则对其进行确证。经确证分析被测物质量色i普降保留时间与标准物质相一致，并且在扣除背景后的样品i苦图中，
9、所选择的离子均出现;同时所选择离子的丰度比与标准样物质相关离子的相对丰度一致(m/z 181、209、211、268，其丰度比34: 100: 65: 55)，相似度在允差之内(见表1)，被确证的样品可判定为氟萃定阳性检出。氛草标准品衍生物的气相色谱质选择离子色谱图和质t曾因参见附录A中图A.l和附录日中图B.l。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/%允许的相对偏差/%6.5 空白试验50 土20除不称取试样外，均按上述步骤进行。7 结果计算和表述20-50 10-20 土25土30用也谱数据处理机或按式(1)计算试样中氟萃定残留量，计算结果应扣除空白值。式中zx 二三二王一
10、hs m x一一试样中氟草定残留盐，单位为毫克每千克(mg/kg); h-一一样液中氟卒定的色谱峰高;h，一一标准工作液巾氟草定的色i苦峰高;C一一标准工作液中氟草定的浓度，单位为微克每毫升(g/mLl; V-佯液最终定容体积.单位为毫升(mL);m一一段终样液所代表的试样质量.单位为克(g)。B 测定低限和回收率8. 1 测定低限和确证低限本方法的测定低限为0.02mg/kg. 8.2 添加浓度范围及回收率本方法添加浓度范围及回收率见表2.lO 土50( 1 ) 3 SN/T 1947-2007 本方法添加浓度及回收率范围样品名称添加浓度范围I(mg/kg)回收率范围/%柑惰0.020-0.
11、200 82.5-97.2 苹果0.020-0.200 81. 5-95. 0 董菜0.020-0.200 80.5-95.8 白菜0.020-0.200 87.5-95.0 玉来0.020-0.200 8. 0-99. 2 糙来0.020-0.200 86.5-97.2 牛肝0.020-0.200 82.5-99.5 鸡肾11 , 1:J -1，:JU 81. 5-99. 8 也4 1洲才J11vtJI-表2/一一附录A(资料性附录)氟草定标准品衍生物的气相色i曹-质谱选择离子色谱图10.91 6000 5000 4000 3000 2000 1000 / 10.00 15.00 川/r25
12、.00 图A.1面衍生物的气后色i曹-质i普选择离子色谱图SN/T 1947-2007 30.00 mm 5 SN/T 1947-2007 25000 20000 15000 10000 5000 。6 59 70得句R民轧中牛羊100 附录B(资料性附录氟革定标准品衍生物的气相色谱，质谱图209 268 181 150 200 250 图B.1氟草定标准晶衍生物的气相色谱质i普图281 299 318327 切。m/z SN/T 1947-2007 Foreword Annex A and Annex B of this standard are informative. This stan
13、dard was proposed by and was under the charge of the certification and Accreditation Ad ministration of the Peoples Republic of China. This standard was drafted by Jilin Entry-Exit Inspection and Quarantine Bureau and Yunnan Entry-Exit Inspection and Quarantine Bureau. This standard was mainly draft
14、ed by Wang Mingtai ,Mu Jun ,Ma Xiaogang ,Song Liguo , Zhou Xiao,Li Aijun , Chang Donghua , Lu Chunmei. This standard is a professional standard for entry-exit inspection and quarantine promulgated for the first time. Note: This English version , a translation from the Chinese text. is only for refer
15、ence. 7 SN!T 1947-2007 Deter,mination of fluroxypr residues in foods for import and export -GC-MS method 1 Scope This standard specifies a method for determination of fiuroxypyr residue in foods for import and ex port by gas chromatography-mass spectrometry (GC-MS1. This standard is applicable to th
16、e de萨市市tlonand confirmii白而fiul，oxypyrresidue in ora吨e，apple , spinach ,cabbage , corn and browrfrice，b。inefiver,and chicken kidey. 2 Prinple 卢/ / _,.,v The test sample is extracted with ethyl aCI1忌，andclear)ed up by gel permeation chromatography ./ / (GPCl and florisil solid phase extracSPE) colun:A
17、fter being esterified , the sample 回lutionis ready for the determination and confrmation by GC-MS smg externrltandard method 民:，-i三二二1 户心当3 Reagents and materials r. l 3.6 Anhydrous sodium sulfate,dried at 650 C for 4 hours,then stored in a sealed container. 3.7 Sodium sulfate aqueous solution,2%. 3
18、.8 Fluroxypyr standard (C, H5 C2 FN2 0, ,CAS. No. 69377-81-7) : Purity二三98%.3.9 Standard stock solution, accurately weigh appropriate amount of fluroxypyr standard and dis solve with acetone. Dilute with acetone to 100g/mL. The solution is stored in a refrigerator at 0C -4C. 8 SN/T 1947-2007 3.10 St
19、andard working solution, Di1ute the standard stock solution with n.hexane to required con. centration. The solution is stored in a refrigerator at ot -4 c. 3.11 Florisil SPE tube ,Florisil.500 mg.6 mL.or equivalent. 4 Apparatuses and equipments 4.1 GC.MS, Equipped with electro.impact source (EIl. 4.
20、2 4.3 Homogenizer. 4.4 4.5 4.6 Concentrate bottle,50 mL an 4.7 Centrifuge. / 5.1.1叫b句J/I .1 . L Take approximately 500 9 of repreSpntative sample. llect Ihe ediblj parts (do not wash with wa. 、也I., .1 ter)and cut into pieces. Crush with a、trusherint。阳fp.Mix torou)hly. Put into clean containers. Seal
21、 、. - , Ii and label them.、飞0;.;_/ 1. 5.1.2 Corn and brown.rice Take approximately 500 9 of representative sample. Smash thoroughly by a pulverizer. Mix thorough. Iy. Put into clean contaners. Seal and label them 5. 1.3 Bovine lve and chicken kidney Take approximately 1 kg of representatve sample. C
22、ollect the edible parts. Crush with a crusher. Mix thoroughly. Put into clean containers. Seal and label them 9 SN/T 1947-2007 5. 2 Storage of test sample The test samples of cereals. fruits and vegetables should be stored at the temperature ranged from 0C to 4 C. Others should be frozen and stored
23、at the temperature below -18C. Certain measures should be taken to prevent contamination of the samples or decomposition of the residues during the sample preparation procedure. 6 Procedure 6. 1 Extraction Weigh ca 20 9 (accurate to O. 1 gl of the test sample into a 250 mL stoppered Erlenmeyer flask
24、. Add 50 mL of ethyl acetate and 20 9 anhydrous sodium sulfa!e. Homogenize for 3 min by a homogenizer. Filter the extract into a 250 mL concentrate bottle under vacuum. Extract the residue with 50 mL of ethyl acetate again. Combine the filtrate into the same concentrate bottle. Condense to nearly dr
25、yness by a rotary evapora!or under water bath at 4OC. Dissolve the residue with 5 mL of cyclohexane-ethyl acetate(l + 1) and wait for cleaning-up. 6.2 Cleanup 6. 2. 1 GPC cleanup 6.2. 1. 1 GPC operating condition a) GPC column:700 mmx25 mm.Bio Beads S.X3.or equivalent; b) Mobile phase ,cyclohexane-e
26、thyl acetate (1 + 1); c) Flow rate ,5. 0 mL/min; d) Injection volume at sample 100p:5. 0 mL; e) Volume of pre.rinsing, 100 mL; f) Eluting volume,260 mL; g) Collecting volume:70 mL-130 mL. 6. 2. 1. 2 GPC cleanup procedure Take 5 mL of the solution above for阳ntyacrding to the condition described as th
27、e s田tion6.2.1.1. Combine the eluates in the fraction collector(70 mL-130 mUand move to one 250 mL concentrate bottle. Condense to nearly dry by a rotary evaporator in a water bath at a temperature below 40 C. 10 S;II/T 1947-2007 Dissolve the residue with 2 mL of n-hexane and wait for SPE cleaning-up
28、_ 6. 2. 2 SPE cleanup Rinse the Florisil SPE column with 6 mL of n-hexane in advance. Transfer the sample solution into the column. Elute with 10 mL of n-hexane-ethyl acetate (7+3). Collect all eluates into a 50 mL con centrate bottle and condense to nearly dry by a rotary evaporator under water bat
29、h at 401巳.Dissolvethe residue and dilute with methanol exactly to 2 mL and wait for esterification. 6.3 Esterification 6. 3. 1 Sample esterification Add 1.5 mL sulphuric acid in a stopered color comparison tube. Esterify for 10 min under water bath at 95C. Cool down. Add 10 mL sodium sulfate solutio
30、n and exact 5 mL n-Hexane simutanously. Shake for 2 min. Take supernatant of n-hexane extract. Centrifugate for 3 min. Then the final sample solu tion is ready for GC-MS determination. 6.3.2 Standard solution esterification Take appropriate volume of standard work solution. Add 2 mL methanol. and es
31、terify as the section 6.3.1 6.4 Determination 6.4. 1 GC-MS operating condition a) Chromatographic column ,HP-1701 silica capillary column.30 m X O. 25 mm (i. d. ) X 0.25 flm film or equivalent; 20 C /min _O ， 5t/min b) Column temperature,50 C (2 min)一一一一一180C(1min)一一一一.270C(18 min); c) Injection por
32、t temperature,280C , d) Interface temperature, 280 C , e) Carrier gas，Helium.purity99. 999% .flow rate 1.0 mL/min, f) Injection volume, 1L; g) Injection mode,Splitless.open the diverting valve after 1.5 min; 11 SN/T 1947-2007 h) Electron ionization mode: 曰:i) lonization energy: 70 eV; jJ Determinati
33、on mode:SIM; k) Selected monitoring ion(m!z): ion for quantification is 209.and ions for qualification are 209. 181.211.and 268: 1) Solvent protection delay:5 min. 6.4.2 GC-MS determination rid oonfirmation 二Relative ionic abundance/% Allowed relative deviation/% :t 30 三三10:t 50 10-20 6.5 81ank test
34、 81ank test will be conducted according to the procedures above without sample addition. 7 Calculation and expression of the result Calculate the content of fluroxypyr residue in the test sample by GC-MS data processor or according to the formula (1). The result of calculation should be deducted wit
35、h blank value. V一-m 户LU-s 7n hH一= X . ( 1 ) 12 where X-the residue content of fluroxypyr in the test sample,mg/kg, h-the peak height of fluroxypyr in the sample solution, hs-the peak height of fluroxypyr in the standard working solution, C the concentration of fluroxypyr in the standard working solu
36、tion，g/mL， V-the final volume of the sample solution ,mL, m-the corresponding mass of the test sample in the final sample solution ,g. 8 Detection limit and recovery 8. 1 Limit of quantification The limit of quantification o this method is 0.02 mg/kg. ificati口nand rec叶SN/T 1947-2007 Sample Fortifica
37、tion range/ (mg/kg) Recovery range/% orange 0.020-0.200 82.5-97.2 apple .,.,-件-0.200. 81.5-95.0 spinach / 产胁。200气80.5-95.8 cabbiage t I 0.020气p:豆o飞87.5-95.0 J I O. 020fO. 200 corn 84.0-99.2 brown-rice ) 0.02-0叩86.5-97.2 bovne liver 飞0.020-0.200 82.5-99.5 , chicken kidney 1 0/ 0J 200.2 lO / 81.5-99.8
38、 F 13 SN/T 1947一2007才6000 5000 4000 3000 2000 1000 14 Annex A (informative) GC-MS selected ion chromatogram of the fluroxypyr derivative 10.91 10.00 15.00 20.00 25.00 Figure A. 1一GC-MSselected ion chromatogram of the fluroxypyr derivative 30.00 mm SN/T 1947一2007Annex B ( informativel GC-MS spectrum
39、of fluroxypyr derivative 209 25000 20000 15000 ISI 10000 5000 236 。4且7，st100 150 200 mlz Figure B. l-GC-MS spectrum of fluroxypyr derivative 中华人民共和国出人境检验检疫行业标准迸出口食品中氟草定残留量的检测方法气相色谱质谱法SN/T 1947-2007 中国标准出版社出版北京复兴门外三里河北街16号邮政编码，100045网址电话，6852394668517548 中国标准出版社秦皇岛印刷厂印刷 开本880X1230 1116 印张1.25 字数26千字2007年11月第一版2007年11月第一次印刷印数1-2o 书号，155066.2-18229 定价12元
