1、中华人民共和国出入境检验检疫行业标准SN/T 1968-2007 进出口食品中扑草净残留量检测方法气相色谱-质谱法Determination of prometryne residues in food for import and export GC-MS Method 2007-08-06发布2008-03-01实施中华人民共和国发布国家质量监督检验检夜总问中华人民共和国出人挠检验检疫行业标准迸出口食品中扑草净残留量检测方法气相色谙质谱法SI/T 1968-2007 号幡中国标准出版社出版北京复兴门外兰里河北街16号邮政编码，100045网址电话，6852394668517548 中国标准
2、出版社秦皇岛印刷厂印刷电坠开本880X12301/1 6 印张1.25 字数33千字2007年II月第一版2007年11月第一次印刷印数1-2000. 书号155066.2-18249定价12.00元SN/T 1968-2007 前言本标准附录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、中华人民共和国福建出人境检验检疫局。本标准主要起草人z陈冬东、李淑娟、李晓娟、蔡;慧霞、安娟、冯帆、李建中、底英章。本标准系首次发布的出人境检验检疫行业标准。I 1 范围进出口食品中扑草净残留量检测方法气相色i昔-质谱
3、法本标准规定了食品中扑草净残留量松洲的制伴和气相色i-1-质i古检测方法。SN/T 1968-2007 本标准适用于大米、花生、胡萝卡、阿兰花、西红柿、洋葱、蘑菇、苹果、柑锅、板栗、鸡肉、牛肉、鸡肾、紫菜中扑草净残留莹的测定和确证。2 方法提要试样中残臼的扑革净IJ乙腊提氨f适合硅胶团中日萃取柱净化.m气相3 试剂和材料3. 1 乙脐z分析纯，色i晋纯。3.2 Fjl l。3. 3 乙酸乙醋。3. 4 丙酬。3. 5 二氯甲烧。3. 6 正己炕3分析纯，包i纯。3.7 氨水:浓度25%。磷股氢二钊J0 3. 15 扑草11标准物质=纯度大于等于99%.英文通用名Cprometrync).CAS
4、 No. 7287-19-6 0 3. 16 扑草净标准溶液，lil确称取适应的扑草净标准物质，用乙脐自己成浓度为100.0mg/L的标准储备溶液，401巳下迎光保在。根据市要用乙肪将储备液稀释成适当浓度的标准工作溶液，现用现配。3. 17 阳离子交换回相萃取柱CSCX), 500 mg.3 mL.戎相当者。3. 18 石罢化碳黑白相萃取柱CEnvi-Carb), 500 mg.6 mL.戎相当者。3.19 N丙基乙二氨键合硅胶因tll萃取柱CPSA),200 mg.3 mL.或相当者c4 仪器和设备4. 1 气有l色i古质谱仪2配有电子轰i:I;电离源CED。4.2 样品粉碎饥。SN/T 1
5、968-2007 4.3 振荡器。4.4 旋转蒸发仪。4.5 pH 计。4.6 氮吹仪。4. 7 固相萃取装置。4.8 涡旋振荡器。5 试样制备和保存5. 1 试样制备5. 1. 1 大米取有代表性样品500g.粉碎并使其全部通过孔径为2.0mm的样品筛。混合均匀，装入沽净的容器内，密封并标识。5. 1. 2 胡萝卡、西兰花、百红柿、洋葱、蘑菇、苹果、柑榻取有代表性样品500g.取可食部分后将其切成小块(不可水洗)，用组织捣碎饥将样品匀浆，混合均匀，装人i古净的容T.内，密封并标识。5. 1. 3 花生、板栗取有代表性样品500g.取可食部分，粉碎并使其全部通过孔径为2.0mm的样品筛。混合均
6、匀，装人沽净的容器内，密封并标识。5. 1. 4 鸡肉、牛肉、鸡肾取有代表性样品500g.切碎并用组织t.%.字机将样品加工成浆状，混合均匀，装入洁净的容器内，密闭并标识。5. 1. 5 紫菜取有代表性样品50g，粉碎，混合均匀.装入洁净的容器内，密封并标识。5.2 试样保存粮谷、坚果类试样在一4C避光保存g水果、蔬菜、肉、内脏等试样在18C避光保存。取样、制样和样品保存过程中，应防止样品受到污染或友生残留物含量的变化。6 测定步骤6. 1 提取6. 1. 1 胡萝卡、西兰花、西红柿、i丰葱、蘑菇、苹果、柑捕、板栗、花生z称取5g试样(精确至O.Olg)于250 mL锥形瓶中，加入30mL乙脂
7、，于振荡器上振荡20min，静宜10min，过滤于250mL分液漏斗。残渣再加入30mL乙脐提取一次，合并两次Ji液。滤液中加入25g氯化伪和60mL磷股盐缓冲液(3. 12).振摇15min，静置分后，弃去水层，乙脑后过元水硫酸纳后，于40C浓缩至干，残渣用2.0mL乙股乙酣十正己;院f昆合溶液(1+1)溶解。6. 1. 2 大米g称取5g试样(衍确至0.01g)于250mL锥形瓶中.加10mL7)(放置15min，加入30mL 乙脯，于振荡器上振荡20min.静TI10 min，过滤并收集滤液于250mL分液漏斗巾。残渣再加入20 mL乙脂提取一次，过滤.合并两次滤液。lJ液中加入25g氮
8、化纳和60mL磷酸盐缓冲液(3.12).振摇15min.静置分层，弃去水层，乙肪层过元水硫酸纳后，于40充MS，quantiliedby external stand-ard method. 气!3 Reagent and materials ter. 3.1 Acetonitrile , HPLC grade. 3.2 Methanol. 3.3 Ethyl acetate. 3.4 Acetone. 3.5 Dichloromethane. 3.6 n-Hexane, HPLC grade 3.7 Ammonia water:25%. 8 SN/T 1968-2007 3.8 Dibasi
9、c potassium phosphateC几HPO，). 3.9 Monopotassium phosphateC KH, PO,). 3.10 Anhydrous sodium sulfate, Ignited at 650t for 4 h.and stored in desiccator 3. 11 Sodium chloride( NaCIl. 3.12 Phosphate buffer (0.5 mol/L.pH 7.0) ,Weigh 52.7 9 of K,HPO, and 30.2日of KH,PO, in 1 L cylinder.add water to dissolve
10、 and make up to the mark.adjust pH=7. 0 by use 2 mol!L NaOH. 3.15 3.17 SCX.SPE ,500 mg.3 mL.or equivalent. 3.18 t n 向VE 在夕-uuh qh und rv otjli mfjt nr o f3.19 PSA.SPE ,200 mg.3 4 4.1 Gas Chromatograph-Mass 4.2 Sample crusher. 4.3 Shaker. 4.4 Rotary evaporator. 4.5 pH阿1eter.4.6 Nitrogen evaporator. 4
11、. 7 SPE vacuum container. 9 川/T1968-2007 4.8 Vortex oscillator. 5 Sample preparation and storage 5. 1 Sample preparation 5.1.1 Rice Re口resenativesample 500 9 grind thoroughly. pass through 2.0 mm sieve. mix thoroughly, then place in clean containers as the test samples by sealed and labeled. 5.1.2 c
12、arrot.broccoli tomato.onion.mushroom.apple.citrus Represenative sample 500 g , cut into small pieces lor edible part , homogenate thoroughly. then place in clean containers as the test samples by sea1ed and labeled. 5. 1.3 peanut castanea mollissima Represenative sample 500日.grind thoroughly lor edi
13、ble part , pass through 2. 0 mm sieve , mix thor oughly. then place in clean containers as the test samplesby sealed and labeled 5.1.4 chicken.beel.chicken kidney Represenative sample 500g. homogenate thoroughly , then place in clean containers as the test sam ples by sealed and labeled. 5.1.5 Laver
14、 Represenative sample 50 g.grind and mix thoroughly. then place in clean containers as the test sam ples by sealed and labeled 5. 2 Sample storage The test samples 01 cereals and nuts should be stored in - 4 t avoiding from the light, the other samples should be stored below -18t .avoiding Irom the
15、light. In the course 01 sampling and sample preparation.precaution must be taken to avoid contamination or any lactors which may cause the change 01 residue content. 10 S:I/T 1968-2007 6 Procedure 6. 1 Extraction 6. 1. 1 carrot. broccoli. tomato .onion. mushroom. apple. citrus. castanea mollissima.
16、peanut. 5 9 (町，curating to 0.01 g) of test sample is weighed and put into a 250 mL conical flask. At the same time. 30 mL of acetonitrile is added into the conical flask. and shaked for 20 min.let standed for 10 min.and filtered into a 250 mL separatory funnel . The extraction is repeated with 30 mL
17、 of acetonitrile. Com bine the supernatants to the funnel. add 25 9 NaCI and 60 mL phosphate buffer (3. 12). shake for 15 min.lay separation and acetonitrile through anhydrous sodium sulfate.and evaporate to nearly dry in a water bath under 40t. The residue is redissolved in 2.0 mL of ethyl acetate-
18、n-hexane( 1 + 4) 6. 1. 2 Rice ,5 9 (accurating to O. 01 g) of test sample is weighed and put into a 250 mL conical flask. 10 mL water is added into the conical flask let standed for 15 min. 30 mL of acetonitrile is add ed into the conical flask and shaked for 20 min .Iet standed for 10 min and filte
19、red into a 250 mL sepa ratory funnel . The extraction is repeated with 20 mL of acetonitrile. Combine the supernatants to the funnel.add 25 9 NaCI and 60 mL phosphate buffer (3. 12) .shake for 15 min. lay separation and acetonitrile through anhydrous sodium sulfate. and evaporate to nearly dry in a
20、water bath under 40C. Then the residue is redissolved in 2. 0 mL of ethyl acetate-n-hexane( 1 + 4). 6. 1.3 chicken. beef. chicken kidney, 5 9 (accurating to O. 01 g) of test sample is weighed and put in to a 50 mL centrifugc tube.30 mL of acetonitrile is added into the centrifuge tube and shaked for
21、 20 min.let standed for 10 min and filtered into a 250 mL separatory funnel. The extraction is repeat ed with 30 mL of acetonitrile. Combine the supernatants to the funel.add 25 9 NaCI and 60 mL phos phate buffer(3. 12) shake for 15 min .Iay separation wipe off water. then purify from fats by extrac
22、 tion with 20 mL acetonitrile saturated hexane. shake for 10 min. wipe off hexane. Acetonitrile through anhydrous sodium sulfate is evaporate to nearly dry in a water bath under 40t . Then the residue is redissolved in 2.0 mL of ethyl acetate-n-hexane(l +4) 6.1.4 Laver ,3 9 (accurating to O. 01 g) o
23、f test sample is weighed and put into a 250 mL conical flask. 10 mL water is added into the conical flasklet standed for 15 min. 30 mL of acetonitrile is add ed into the conical flask and shaked for 20 min. let standed 10 min and filtered into a 250 mL separato ry funnel The extraction is repeated w
24、ith 20 mL of acetonitrile. Combine the supernatants to the funnel.add 25 9 NaCI and 60 mL phosphate buffer (3.12) .shake 15 min.lay separation. And acetoni trile through anhydrous sodium sulfate is evaporated to nearly dry in a water bath under 40t . Then the residue is redissolved in 2.0 mL of ethy
25、l acetate-n-hexane(l +4). 6.2 Clean up 6.2.1 Extraction of sample from 6. 1. 1 .6. 1.2 and 6. 1.3 is cleaned up by Envi-Carb and PSA. Fiest. V飞thichare washed with 10 mL and 4 mL ethyl acetate-n-hexane( 1 + 4). tandem 2 column( Envi-Carb 11 SN/T 1968-2007 u口.PSA down). Pass the sample extract soluti
26、on through the tandem column at a rate of2 mVmin Rinse the column with 3 x 2 mL ethyl acetate-n-hexane( 1 + 4Collect elution in a tube. Nitrogen evaporator to dry in a water bath under 40t .add 1.0 mL acetonitrile and ready for GC-MS determi-nation. 6. 2. 2 Extraction of sample from 6. 1.4 is cleane
27、d up by Envi-Carb and SCX. Envi-Carb is washed with 10 mL ethyl acetate-n-hexane (1 +4). Pass the sample extract solution through the column at a rate of 2 mLlmin. Rinse the column with 3 x 2 mL ethyl acetate-n-hexane(l + 4). Collect elution in a tube. Nitrogen evaporator to dryness. add 1 mL n-hexa
28、ne solution. SCX is activated with 2 mL acetone and 2 mL dichloromethane. pass the solution through the column. Rinse the lumn with 2 x 1 mL n-hexane. 2 mL dichloromethane. 2 x 2 mL acetone in turn. elution by 2 mL ammoniated methanol(3.14). Collect elu山tionin fs盹e口尚同忡al玩r肖泊=玩ator叩f扣Lmnel，l巾叫白民飞dd1口
29、mLp仙11咀咀!$pl灿hat白ebu时ff怡e町r叫 12) and shake J .;吃二jU队Reextractthrice with 10 mL.1QmL.5_JnL己d怕10romethane.Dichlcjromethanethrough anhydrous t飞山lsodium sulfate. Nitrogen evaporatto drvrie白.ad!L1:0 mL acetonitrjle and ready for GC-MS deter-mination. ./ _ ;/ 6.3 Determination 6.3.1 GC-MS operating condit
30、ionsj a) Column , DB-5MS.30 m x O. 25 b) Column temperature,70t (1 min c) Carrier gas , He. Purity ;:99. 99!f% ; d) Injection temperature,260t ; e) Flow r8te,1.0 mL/min; f) Injection mode, Splitless; g) Injection volume,l. 0L; h) lon source ,EI.70 eV; i) Source temperature,230t; j) Ouadrupole temper
31、ature,150t; k) Interface temperature,280t; 12 , ;/ / 一队一jeiin1BGh巴280C(3min); S:-I/T 1968-2007 1) Selected ions(m/z) ,241.184.226.199. 6.3.2 GC-MS determination According to the approximate concentrat旧n01 prometryne in sample solution. select the standard working solution with similar peak area to t
32、hat 01 sample solution. The responses 01 the analyte in the standard working solution and the sample solution should be within the linear range 01 the instru ment detection. The retention time 01 prometryne is about 11.5 min under the above conditions. The Choromatogram 01 total ion current 01 the s
33、tandard. see annex A. Qualitative ion of prometryne is 241.184.226.199. The quantitation ion 旧1日4.For the same analysis batch and the same mpound.thefvariation range 01 the ion ratio at tre similar concentration can not f range 1川川W叫t仙阳阳he阳e凹n巾m叫po阳川口m州n时dl步功dQualitative V Compound Qualitative ion (
34、m/z) / Stand 凹飞ratio/%Permitted to!erances/% 241.00 / YY /yJ10O 184/ 可言10 Prometryne 226.00 10 199.00 15 -6. 4 81ank test 川口阳阳阳e町era阳a) f. / 勺川.7 Ca创Ic蛐u川由|阳a挝甜削叫ti阳llona川nd臼叫叩pr阳叩e邸s岱s臼蚓叫叫s创叫呻呻i呻咿耻+如n刊of加t归阳阳阳阳e臼阻削叫叫s缸叫圳u川山川圳i忱忖t1 . . 1 1 . I T阳h怕阳ecoAJcsVV叫J忡I/ 一-一一一.，:_L- ( 1 ) As x m where X-the
35、 residue content of prometryne in test sample.mg/阔，A -peak area 01 prometryne in sample s口lution;As-peak area of prometryne in standard solution; Cs -concentration 01 prometryne in standard solution.g/mL; 13 SN/T 1968-2007 V一thefinal volume of the sample solution.mL, m-mass of test sample,g. NOt6: T
36、he blank value shall be subtracted from the above resut of cakulation 8 Limit of determination and recovery 8. 1 Limit of determination The limit of determination of this method is 0.01 mg/kg. 8.2 Add concentration and recovery Data are shown in Table 2. Table 2-Add concentration and recovery Sample
37、 Add concentration, (mg Ikg) Recovery/% 0.01 102.0日-109.76nce 0.02 98.18-109.47 0.05 101.74-108.78 0.01 82.23-95.34 peanut 0.02 90.16-102.71 0.05 86.73-95.86 0.01 74.19-87.12 carrot 0.02 70.21-90.12 0.05 82.73-88.30 0.01 90.74-115.38 broccoli 0.02 96.44-109.8 0.05 84.08-108.86 0.01 75.54-87.44 to口18
38、tO0.02 86.67-95.00 0.05 85口4-91.990.01 74.54-86.83 mushroom 0.02 82. 67 - 96. 88 0.05 85.04-91.38 0.01 77.55-96.05 onlon 0.02 86.44-104.62 0.05 88.53-102.24 14 SN/T 1968-2007 Table 2( Continued) Sample Add concentrationl(mg/kg) Recovery/% 0.01 81.40-102.17 apple 0.02 82.52-100.83 0.05 86.85-102.20 0
39、.01 82.36-104.18 citrus 0.02 85.83-108.60 。.0582.16-104.79 0.01 74.19-88.20 castanea mollissima 0.02 85. 04 - 96. 26 0.05 80.16-100.18 0.01 81.62-97.62 chicken 0.02 83.54-97.61 0.05 88.58-102.36 0.01 82. 16-102. 4B beet 0.02 86.80-108.33 0.05 92.58-107.13 0.01 88.40-102日4chicken kidney 。.0284.68-102
40、.55 0.05 79.73-87.80 0.01 75. 54-87. 44 laver 0.02 86. 67 - 93. 94 0.05 85.73-91.99 SN/T 1968-2007 Annex A (informative annex) Choromatogram of total ion current TIC) and mass spectrum Abundance 80000 75 000 70 000 65 000 60 000 55 000 50 000 45 0日040 000 35 000 30 000 25 000 20 000 15 000 10 000 TI
41、C:6122108_D飞data.ms11. 546 J于一_.JL-一卢卢忖JF8.50 9.00 9.50 10.00 10.5011. 00 11.5012.0012.5013.00 13.5014.0014.5015.00 Time一一-Figure A. l-Choromatogram of total ion current of prometryne standard Abundance 9 000 8 500 8 000 7 500 7 000 6 500 6000 5500 5000 4 500 4 000 3 500 3 000 2500j 58 2 000 1 500 1 000 阳r时11川11.5川n:612川Ddilta，ms-)! 241 184 210 t t 261280300 3213423623R3 406 430 4阳的1500 。阳100150白o250 3皿350400 4民500、，-Figure A. 2-Mass spect阳mof prometryne standard 书号，1550662-18249 定价，12.元
