1、S 中华人民共和国出入境检验检疫行业标准SN/T 1990-2007 进出口食品中三瞌锡和三环锡残留量的检测方法气相色谱-质谱法Determination of azocyclotin and cyhexatin residues in food for import and export-GC-MS method 2007-08-06发布2008-03-01实施护黑头中华人民共和国发布日黯中国家质量监督检验检班总川吧7中华人民共和国出入境检验检疫行业标准进出口食品中三睡锡和三环锡残留量的检测方法气相色谱质谱法SJ/T 1990-2007 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
2、，100045网址WW屯话:685239668517548 中国标准出版社奉皇岛印刷厂印刷开本880X12301/16 印张1.25 字数32千字2007年11月第一版2007年11月第次印刷印数1-2000陪书号:155066.2-18275 定价12.00元前言本标准的附录A为规范性附录、附录B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。S:-;jT 1990-2007 本标准由中华人民共和国陕西出入境检验检疫局、中华人民共和国天浮出人统检验检疫局、中华人民共和国内蒙古出入境检验检疫局、中华人民共和国山东出入境检验检疫局、中华人民共和国柄建出入境检验检疫局负责起草。本标准主
3、要起草人:李建华、何强、孔祥虹、乐爱山、葛宝坤、林150 20-50 10-20 10 GCMS时相对离子丰度品大允许误差/tIO _j_ 15 噜-20土506.6 2白实验除不加试样外，均按上述测定步骤进行。7 结果计算和表述测定结果以三环锡L若以三l呢锡计时将二环锡含且1乘以换算系数1.13. 用色1市数据处理机或按式()计算试样中三环锡的含妇.计算结果须扣除空白的。V一-m F叫一-A A一X . ( 1 ) 3 SN/T 1990-2007 式中zX 试样中三环锡的含量，单位为毫克每千克(mg/kg); A样液中三环锡衍生物三环己基甲基锡的色谱曲革面积;Cs一一标准工作液中三环锡的浓
4、度，单位为微克每毫升(g/mU;V 样液最终定容体积，单位为毫升(mL), A民标准工作液中三环锡衍生物三环己基甲基锡的色i吉峰面帜$111一最终样液所代表的试样量，单位为克(g)0 8 测定低限、回收率8. 1 m定低限本方法的测定低限为O.。8.2 回收率样品的添加浓度及回收率g实验数据见表20样品添加浓度I(mg/kg)回收率范eyf，样几/1添加浓度I(mg/kg)回收率范围1%0.020 78/97C2rt酣0.020 76.3-101. 9 茶叶0.040 /s9.Q-07, 6 0.040 84.1-104.5 。080今-85-:_5;:10i).1 0.080 85.4-10
5、2.9 0.020 92.:)-引10.020 78.4-97.0 韭菜。.040.!LJ， ! 一世拚主革0.040 89.0-97.6 0.080 87. -102. 3 . 飞、0.080 85.5-100.4 0.020 A100.8/ 0.020 83.5-96.1 苹果0.040 11 Sd-99. 0.040 84.2-96.4 0.080 1: 81，沪10211r飞-、0.080 85.4-98.9 0.020 1气rf.6-99/10.020 81. 1-97.9 板栗0.040 屯4-1才3鸡肉0.040 83.5-98.1 0.080 76)-胃号飞亿3L 。08083
6、.5-100.3 。.020mhoqt/ll / 0.020 82.3-9 1. 9 甘草0.040 X!等.ll号15/宣巳J/ 0.040 76.8-94.9 0.080 90.6-100.0 0.080 77.3-97.9 0.020 71. 5-96. 7 0.020 71. 3-99. 0 大米0.040 76.0-103.9 猪肝0.040 77.1-103.6 0.080 84.2-102.1 0.080 80.7-100.4 4 附录A(规范性附录)甲基破化援溶液的合成方法A.1 试剂和材料A. 1. 1 无水乙能:用金属纳脱水。A. 1. 2 石油酿沸程60C90C .用金属
7、纳脱水。A.1.3 fR带:60C干燥30mino A. 1.4 矶甲:院:重蒸饱。A.2 A. 2. 3 A. 2. 4 A.2.5 A.3 合成/ SN/T 1990-2007 5 SN/T 1990-2007 附录B(资料性附录)三瞠锡和三环锡标准物质衍生物三环己基甲基锡的选择离子色i曹图、质谱图Abundancc 5000 -4000 -3000 -2000 -1000 -k Abundance 80000 60000 55 40000 41 20000 。6 10.77 10.00 -图B.1三环已基甲基锡的选择离子色i曹图219 301 135 9 1 97 111. 161 17
8、5 241 269 287 III 1 100 200 300 图B.2三环己基甲基锡的全扫描质i曹图15.00 I/mn 369 384 m/z SN;T 1990-2007 Foreword Annex A of this standard is normative annex and annex B is informative annex. This standard is proposed and is under the charge of the Certification and Accreditation Administra tion of the Peoples Repu
9、blic of China This standard was drafted by the Shaanxi Entry.Exit Inspection and Ouarantine Bureau of the Pe。口lesRepublic of Chi阳，Tianjin Entry-Exit Inspection and Ouarantine Bureau of the Peoples Republic of China , Neimenggu Entry-Exit Inspection and Ouarantine Bureau of the Peoples Republic of Ch
10、ina , Shandong Entry-Exit Inspection and Ouarantine Bureau of the Peoples Republic of China and Fujian Entry.Exit Inspection and Ouarantine Bureau of the Peoples Republic of China. The principal drafters of this standard are Li Jianhua , He Oiang , Kong Xianghong , Yue Aishan , Ge Baokun , Lin Anqin
11、g , Li Gang , Pan Guoqin日，Wang Jianhua , Yang Fang This standard is the first special normal standard of the Entry-Exit Inspection and Ouarantine of the Peoples Republic of China Note , Th旧Englishvers旧n.a translatio、fromthe Chinese text. is solelfor guidance 7 S;I/T 1990-2007 Determination of azocyc
12、lotin and cyhexatin residues in food for import and export-GC-MS method 1 Scope This standard specifies the method for the determination and confirmation of azocyclotin and cy hexatin residues by gas chromatography-mass spectrometry in food. This standard is appli臼bleto t嗜detem、nationand confirmatio
13、n f residue contents of azocyclotin and cyhexatin in tea , leek , appl, chestnut. glycyrrhizae , rice , vijlegar , honey. beef, chicken , fish and liver.凰2 Principle 3 Reagents and materials . and water is distilled water. 3.2 3.3 3.4 Methyl magnesium iodide solution: synthesized according to annex
14、A 3.5 Hydrobromic acid. 3.6 Anhydrous sodium sulfate: Ignite at 650巳for4 h , and keep in a desiccator. 3.7 Hydrochloric acid solution: 10%. 3.8 Sodium chloride solution: 20 9 I L. 8 SN/T 1990-2007 3.9 Azocyclotin standard (C20H35N,Sn ,CAS No ,41083-11-8) , Purity;?98%. 3.10 Cyhexatin standard (C18H,
15、.OSn ,CAS No , 13121-70-5) , Purity;?98%. 3. 11 Standard stock solution, Accurately weigh an adequate amount of azocyclotin standard and cyhexatin standard and dissolve in a small volume acetone respectively. Dilute with acetone to form a standard stock solution of 100g/mL in concentration. Store at
16、 4t 3.12 Standard working solution, Then dilute the standard stock solution with acetone to the re quired concentration as the standard working solution. Store at 4t. 3.13 Florisil cartridge, 1 g, mL. 4 Apparatus and equipmnt 4. 1 4.2 Tissue blender. 4.3 Grinder. 4.4 Homogenizer. 4.5 Shaker. 4.6 Rot
17、ary vacuum evapor 4.7 Solid phase extraction 4.8 Centrifuge. 4.9 Centrifuge tube, 50 mL. / ass detectqpff / f / 5 Preparation and storage of test samples 5. 1 Preparation of test samples 5. 1. 1 Fruits, nut and vegetables The combined prima叩samplesare reduced to the ca 500日，which has been removed sh
18、ell. seed. peel , stem. root. coronal (do not wash by water). The edible portions are cut and homogenized thoroughly in a high speed blender. Keep the prepared sample into a clean container. sealed and la beled 9 SN/T 1990-2007 5.1.2 Tea and glycyrrhizae and grains or cereals The combined primary sa
19、mples are reduced to the ca 500日，which is crushed with a grinder and let wholly pass through 2.0 mm sieve. Kee口theprepared sample into a clean container, sealed and la beled. 5. 1. 3 Meats and meat products The combined primary samples is reduced to ca 500 g , the edible potions are thoroughly groun
20、d and homogenized in a meat grinder. Keep the prepared sample into a clean container , sealed and labeled. 5.1.4 Honey Take about 500 9 01 representative sample. The sample that is not crystallized shall be stirred well to produce a homogenous sample. 1I the sample is crystallized , it should be war
21、med in a waterbath at below 60C with the sample bottle covered tightly. Mix thoroughly when all sample has melted. then cool immediately to room temperature. In the course 01 melting the sample. precautious measures must be taken to avoid evaporation of water from the sample. Keep the prepared sampl
22、e into a clean container, sealed and labeled. 5. 2 Storage of test samples The test samples 01 tea. bee products. Chinese herbs, grains or cereals should be stored below 4l:. The test samples of fresh fruits. vegetabl田，meat and meat products should be stored below -18C In the course of sampling and
23、sample preparation. precaution should taken to avoid contamination or any factors which may cause the change of residue content. 6 Method of determination 6. 1 Extraction For solid samp怡.such as tea. leek. apple, chestnut. glycyrrhizae. rice. beef. chicken. fish and liv 町，weigh 5 gCaccurate to O. 01
24、g) of the test sample into a 50 mL centrifuge tube. Add 10 mL water and 5 mL hydrobromic acid , stand for 2 h after shake 30 s drastically. Add 20 mL acetone. extract for 5 min on a high speed homogenizer , and centrifuge for 3 min under 3 000 r/min. Transfer the up per extract solution into a 150 m
25、L concentrating bottle by passing through anhydrous sodium sulfate column. Repeat above extract procedure with 20 mL of acetone twice. Combine acetone extracts, evaporate to 20 mL in a rotary evaporator with a bath temperature below 350C. Add 20 mL NaCI so lutionC3. 8) and 2 x 40 mL petroleum ether,
26、 shake 5 min , transfer the upper organic phase into anoth er 150 mL concentrating bottle through anhydrous sodium sulfate column. Add another 2 x 40 mL pe-10 S:-I/T 1990-2007 troleum ether and shake 5 min , discard the lower aqueous lay凹，combine the organic phase and evaporate to approach dryness i
27、n a rotary evaporator with a bath temperature below 40C and blow to dryness. Dissolve the residue with 20 mL anhydrous ether (3.3) For liquid sample such as vinegar and honey, weigh 5 g(accurate to O. 01日)of the test sample into a 150 mL concentrating bottle. Add 20 mL water, 5 mL hydrobromic acid ,
28、 10 mL acetone and shake 10 min. Extract for 5 min with 2 x 30 mL petroleum ether twice by shaking. Transfer the upper or ganic phase into another 150 mL concentrating bottle by passing through anhydrous sodium sulfate column. Evaporate to approach dryness in a rotary evaporator with a bath temperat
29、ure below 40 C and blow to dryness. Dissolve the residue with 20 mL anhydrous ether(3. 3). 6.2 Derivation Add 5 mL methyl magnesium iodide solution(3. 4) into above anhydrous ether solutionC6. 1) , shake 5 min , then add 10 mL 10% hydrochloric acid solution(3. 7) and shake 2 min. Stand for separate
30、into two layers and discard the lower aqueous layer. Evaporate to dryness in a rotary evaporator with a bath temperature below 35C. Dissolve the residue with 10 mL petroleum ether. 6.3 Clean up Set up the solid phase extraction vacuum manifold and mechanical pumpCabout 1 cm thickness anhy drous sodi
31、um sulfate was put into the florisil cartridge). Rinse the cartridge with 5 mL petroleum ether and then transfer the above solution C 6. 2) into cartridge. Elute with 15 mL petroleum ether with the flow rate below 3 mL/min. Collect all the eluted solution in a 150 mL concentrating bottle and evapora
32、te to approach dryness in a rotary evaporator with a bath temperature below 40 C and blow dryness with nitrogen. Dissolve the residue and dilute exactly to 1. 0 mL with acetone for GC/MSD. For complex test sample matrix , such as tea , leek , glycyrrhizae , and liver, cleaning up operate with two fl
33、orisil cartridges connecting in series 6.4 Prepare standard solution Transfer 1.0 mL cyhexatin standard working solution in appropriate concentration into a 150 mL con centrating bottle. Blow out acetone with nitrogen and dissolve the residue with 20 mL anhydrous etherC3.3) The following operat旧npro
34、cedures are same as that of test sample with derivate (6.2) and clean up(6. 3) 11 SN/T 1990-2007 6. 5 Oetermination 6.5. 1 GC/MSO operating conditions a) Column: 08-5 ms fused q国内Z田口illarycolumn. 30 m X O. 25 mm (id). film thickness O. 25m or the equivalent; 25t/min _ 25t/min b) Column temperature:
35、160C (0 min)一一一一205C(10 min)一一260C(5 min) ; c) Injection port temperature: 230 C ; f) Injection volume: 1L; g) Injection mode: Splitless. purg h) lonization mode: EI; j) lonization energy: 70 eV; j) Acquistion mode: SIM; k) Monitor ion (m/z): 121. 135. 1) Sovlent delay: 6 min. 6.5.2 According to the
36、 approximate concentration of tricyclohexylmethyltin in the test sample solution. select the standard working solution with similar peak area to that of sample solution. The standard working solution should be injected randomlv in between the injections of sample solution of equal volume. The respon
37、ses of tricyclohexylmethyltin in the standard working solution and sample solu tion should be in the linear range of the instrumental detection. According to the GC/MSO operating conditions (6.5.1). if the retention time of sample chromato gram peaks are consistent with the standards. and subtracted
38、 from background compensat旧n.se lected ions are all present and the relative ion abundance of the selected ions according with that of the calibration standard. at comparable concentrations. within the tolerances (seen table 1). Under 12 SN/T 1990-2007 the above GC/MSD operating conditions , the ret
39、ention time oftricyclohexylmethyltin is 10. 8 min , and the ratio of the monitoring ions (m!z) is 121 : 135 : 219 : 301 = 25 : 57 : 100 : 44. For GC-MS chromatogram (TIC) and mass spectrum of the standard , see figure B. 1 and B.2 in annex B. Table l-Maximum permitted tolerances for relative ion abu
40、ndance while confirmation Re!ative abundance 50 20-50 10-20 10 (base peak)/% Permitted 士10:t 15 士20士50tolerances/ % a d山-_. 1 ( . . . . . . . . . . . . . . . . . . . . . . . . . . . where X -the residue content of azocyclotin 叫/m s A-the peak area of tricyclohexylmethyltin in the sample solution; cs
41、-the concentration of azocyclotin or cyhexatin in the standard working solution，g/mL; V-the final volume of the sample solution, mL; As-the peak area of tricyclohexylmethyltin in the standard working solution; m一thecorresponding mass of the test saample in the final sample solution. g. 13 SNjT 1990-
42、2007 8 Limit of determination and recovery 8. 1 Limit of determination The limit of determnaton of ths method is 0.020 mg Ikg. 8. 2 Recovery Fortfying concentrations n test samples and recovery experimental data are I st in table 2. Table 2-Fortifying concentratons in test samples and recovery exper
43、imental data Sample Fortifying Range 01 Fortifying Range of concentration/ (町咱/kg)recove叩/%concentration/ (mg/kg) recovery/% 0.020 78.4-97.0 0.020 76.3-101.9 tea 0.040 89.0-97.6 0.040 84.1-104.5 0.080 85.5-100.4 0.080 85.4-102.9 0.020 92.5-99.1 0.020 78.4-97.0 leek 0.040 91.4-99.4 hone 0.040 89.0-97
44、.6 0.080 87.口-102.30.080 85.5-100.4 0.020 81.5-100.8 0.020 83.5-96.1 apple 0.04口85.2-99.2 beef 0.040 84.2-96.4 0.080 84.5-102.1 0.080 85.4-98.9 0.020 72.6-99.1 0.020 81.1-97.9 chestnut 0.040 84.4-1.3 chicken 0.040 83.5-98.1 0.080 76.9-100.3 0.080 83.5-100.3 0.020 80.7-99.4 0.020 82.3-94.9 glycyrrhiz
45、ae 0.040 86.1-101.8 fish 。口4076.8-94.9 0.080 90.6-1日O.。0.080 77.3-97.9 0.020 71.5-96.7 0.020 71.3-99.0 rice 0.040 76.0-103.9 liver 0.040 77.1-103.6 0.080 84.2-102.1 0.080 80.7-100.4 14 SN/T 1990-2007 Annex A ( normative) Synthesis of methyl magnesium iodide solution A. 1 Reagents and materials A. 1.
46、 1 Anhydrous ether: dehydrate with sodium. A. 1. 2 Petroleum ether: boiling point from 60t to 90t. dehydrate with sodium. A. 1.3 Magnesium ribbon: dried 30 min at 60C. A. 1.4 lodomethane: redist川人1.5lodomethane-anhydrous: 71 9 iodomethane and 400 mL anhydrous ether ether mixture solu tion. A. 1.6 An
47、hydrous sodium sulfate: Ignite at 650t for 4 h. and keep in a desiccator. A.2 Apparatus and equipment A. 2. 1 Heater sheath: 1 000 mL. A. 2. 2 Double neck flask: 1 000 mL. A. 2. 3 Spherical condenser: 400 mm. A. 2. 4 Isobarically funnel: 250 mL A. 2. 5 Dryness tube. A.3 Synthesis Cut the dried magne
48、sium ribbon into 2 mm-3 mm chips. weigh ca 14 9 of magnesium chips into a 1 000 mLdouble necks flask. add 100 mL petroleum ether (A. 1. 2). Set up spherical condenser (with dryness tube packed anhydrous calcium chloride) and isobarically funnel. inlet cooling water into condenser. and heat slowly. T
49、ransfer iodomethane-anhydrous ether mixture solution (A. 1.5) into the funnel. add 60 mL mixture solution (A. 1.5) into the flask first and add the other mixture solu tion during 1 h. Keep the reaction in fluxing 1. 5 h. Transfer the methyl magnesium iodide solution into a brown bottle after cooling to room temperature and place in a desiccator. -SN/T 1990-2007 Annex B (
