SN T 1902-2007 水果蔬菜中吡虫啉、吡虫清残留量的测定 高效液相色谱法.pdf

1、版权所有 禁止翻制、电子传阅、发售中华人民共和国出入境检验检疫行业标准SN/T 1902-2007 水果蔬菜中毗虫日林、日tt虫清残留量的测定高效液相色谱法2007-05-23发布Determination of imidacloprid and acetamiprid residues in fruits and vegetables-HPLC 2007-12-01实施中华人民共和国发布国家质量监督检验检度总局版权所有 禁止翻制、电子传阅、发售目IJ1=1 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。本标准由中华人民共和国江苏出入境检验检疫局负责起草。本标准主要

2、起草人:何松涛、俞畔、乙小娟、张爱东、刘一军、孙建刚。本标准系首次发布的检验检疫行业标准。SN/T 1902-2007 版权所有 禁止翻制、电子传阅、发售1 范围水果蔬菜中毗虫琳、口比虫清残留量的测定高效液相色谱法本标准规定了水果蔬菜中毗虫琳、口比虫清残留量的高效液相色谱测定方法。本标准适用于番茄、黄瓜、柑情中口比虫琳、口比虫清残留量的检验。2 测定方法2.1 试样制备SN/T 1902-2007 将所取原始样品缩分出1kg，取可食部分，经组织捣碎，均分成两份，装入洁净容器内，作为试样，密封，并标明标记。2.2 试样保存将试样于180C以下冷冻保存。注.在试样制备过程中，应防止样品受到污染或发

3、生残留物含量的变化。2.3 方法提要试样中残留的口比虫叫中、H比虫清用乙睛提取，提取液经弗罗里砂柱净化，浓缩、定容后，用配有紫外检测器或二极管阵列检测器的高效液相色谱仪测定。2.4 试剂和材料除另有规定外，所用试剂均为分析纯，水为蒸馆水或去离子水。2.4. 1 乙睛:HPLC级。2.4.2 正己院。2.4.3 丙酣。2.4.4 氧化铀:1400C烘烤4ho 2.4.5 日比虫琳标准品:纯度大于等于96%。2.4.6 日比虫清标准品:纯度大于等于96%。2.4.7 日比虫啡、毗虫清标准溶液:准确称取适量的口比虫啡、H比虫清标准品，用乙腊配制成100g/mL的单个标准储备液，根据需要再用乙睛十水(

4、30十70)稀释成适当浓度的白合标准工作、溶液。2.4.8 固相萃取柱，弗罗里砂柱(Florisil)，体积3mL小柱，填充物500mg o 2.5 仪器设备2.5.1 高效液相色谱仪:配有紫外检测器或二极管阵列检测器。2.5.2 振荡器。2.5.3 氮吹仪。2.5.4 旋转蒸发仪。2.5.5 游涡混合器。2.5.6 微量注射器:100L。2.6 测定步骤2.6. 1 提取称取25g试样(精确至0.01g)，置于250mL具塞锥形瓶中，准确加入50.0mL乙睛，放置2h，振荡提取30min后用滤纸过滤。滤液收集到装有5g7 g氧化铀的100mL具塞量筒中，盖上塞子，剧版权所有 禁止翻制、电子传

5、阅、发售SN/T 1902-2007 烈振荡1mi口，在室温下静止10mi口，使乙睛相和水相分层。从100mL具塞量筒中吸取10.0mL乙腊溶液，移入125mL梨形瓶中，于500C水浴中旋转浓缩至近干。加入10mL丙酬+正己皖(10十90)溶解。2.6.2 净化将弗罗里面夕柱用2mL lE己皖预淋洗，当溶剂液面到达柱吸附层表面时，立即倒入样品溶液，弃去流出液。用5mL丙酬十正己皖(10十90)涮洗梨形瓶后淋洗弗罗里面夕柱，弃去流出液。用10mL丙酣十正己皖(20十80)分两次淋洗弗罗里砂柱，用15mL刻度离心管接收洗脱液。将盛有洗脱液的离心管置于氮吹仪上，在500C水浴下，氮吹至近干，用乙腊+

6、水(30+70)定容至2.0mL，在游涡混合器上泪匀，经O.45m滤膜过滤后，待测。2.6.3 测定2.6.3. 1 液中目色谱参考条件a) 色谱柱:ODS-C18，5m，250mmX 4 mm(内径)或相当柱;b) 色谱柱温度:400C;c) 流动相:乙睛十水，梯度:0mi口，(5十95);15 min, (25十75);20mi口，(5十95);d) 流速:1.0 mL/min; 巳)检测波长:258口m;f) 进样量:20L。2.6.3.2 色谱测定根据样液中被测农药残留限量要求，选定浓度相近的白合标准工作溶液。混合标准工作榕液和待测液中农药的含量在仪器检测的线性范围内，按上述色谱条件进

7、行测定，口比虫琳和口比虫清的保留时间约为13min和15mi口。标准品的液相色谱图参见附录A。2.6.4 空白试验除不加试样外，按上述测定步骤进行。2. 7 结果的计算和表达按式(1)计算试样中毗虫琳、口比虫清的残留量，计算结果需将空白值扣除。式中:x 生兰主兰主A , Xm X 试样中口比虫叫中、H比虫清含量，单位为毫克每千克(mg/kg); A 试样中口比虫啡、H比虫清的峰面积;c 标准工作液中毗虫啡、毗虫清的浓度，单位为微克每毫升(g/mL); V 样液最终定容体积，单位为毫升(mL);A 标准工作液中毗虫啡、毗虫清的峰面积;m 最终样液所代表的试样量，单位为克(g)o3 测定低限、回收

8、率3.1 测定低限本方法测定低限为0.02mg/kg o 3.2 回收率3.2.1 番茄中毗虫琳、毗虫洁的添加浓度及其回收率的实验数据:. ( 1 ) 添加浓度在0.02mg/同时，毗虫琳回收率为91.4%109%，毗虫清回收率为95.5% 108% ; 添加浓度在0.05mg/kg时，毗虫琳回收率为89.2%101%，毗虫清回收率为90.O% 2 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 102% ; 添加浓度在o.10 mg/kg时，毗虫琳回收率为80.7% 88. 5%，毗虫请回收率为85.5% 103%。3.2.2 黄瓜中毗虫琳、口比虫清的添加浓度及其回收率的实验

9、数据:添加浓度在0.02mg/同时，毗虫琳回收率为98.5%106%，毗虫清回收率为94.O% 106% ; 添加浓度在0.05mg/同时，毗虫琳回收率为95.0%106%，毗虫清回收率为90.O% 102% ; 添加浓度在o.10 mg/同时，毗虫琳回收率为81.4%97. 2%，P比虫清回收率为96.2% 109%。3.2.3 柑情中毗虫琳、口比虫洁的添加浓度及其回收率的实验数据:添加浓度在0.02mg/kg时，毗虫琳回收率为94.5% 102%，毗虫清回收率为90.0% 99.0% ; 添加浓度在0.05mg/kg时，毗虫琳回收率为88.0%98.4%，毗虫清回收率为84.4% 93.8

10、% ; 添加浓度在O.10 mg/kg时，毗虫清回收率为84.9%109%，毗虫清回收率为87.3% 99.5%。3 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 附录(资料性附录)标准品高效液相色谱图A 但如司运-ECUZ垂4叫世=EENmMJ 0.25 。0.2岳o.岳。75-_-. 一二)一一一一1. 25 lm 17 16 1岳14 13 12 11 10 9 8 1. 5 标准品液中目色谱固(0.08g/mL)固.14 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 Foreword Annex A of this standard is an in

11、formative one. This standard was proposed band is under the charge of National RegulatorCommission for Certi fication and Accreditation. This standard was drafted by Jiangsu Entry-Exit Inspection and Quarantine Bureau of the People s Republic of China. The main drafters of this standard are He songt

12、ao, Yu ye , Yi Xiaojuan , Zhang aidong , Li u yijun , Sun jlangang. This standard is an inspection and quarantine professional standard promulgated for the first time. Note: This English version.a translation from the Chinese text. is solelfor guidance. 5 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 Determinatio

13、n of imidacloprid and acetamiprid residues in fruits and vegetables -HPLC 1 Scope This standard specifies the method of determination of imidacloprid and acetamiprid residues in fruits and vegetables by HPLC. This standard is applicable to the determination of imidacloprid residue in tomatoes , cucu

14、mbers and oranges. 2 Method of determination 2. 1 Preparation of test sample The combined primary sample is reduced to 1闸，theedible portions are blended , and divided into two equal portions. Each portion is placed in a clean container as the test sample , which is then sealed and labeled. 2. 2 Stor

15、age of sample The test sample should be stored below -1SC. Note: In the course of sample preparation , precautions must be taken to avoid contamination or any factors that may cause the change of residue content. 2.3 Principle The imidacloprid and acetamiprid residues in the test sample are extracte

16、d with acetonitrile. The ex tract is cleaned up by Florisil SPE tube and evaporated. The residue is dissolved and made up to a definite volume. Determination is made by means of a HPLC with UV detector or DAD. 2. 4 Reagents and materials Unless otherwise specified. all the reagents used should be an

17、alytically pure. Water is distilled water or de-ionized water, 2.4, 1 Acetonit门le:HPLC grade. 6 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 2.4.2 N-hexane. 2. 4. 3 Acetone. 2.4.4 sodium chloride: baked at 140C for 4 h. 2.4.5 Imidacloprid standard: Purity no less than 96%. 2.4.6 Acetamiprid standard:Purity no le

18、ss than 96%. 2.4.7 Imidacloprid and acetamiprid standard solution: Accurately weigh an appropriate amount of imidacloprid or acetamiprid standard ,dissolve in acetonitrile to prepare a single standard stock solu tion of 100g/mL. Dilute the standard stock solution with acetonitrile-water (30 + 70) to

19、 the re quired concentration as the mixed standard working solution. 2.4.8 SPE tube, Florisil ,3 mL,500 mg. 2.5 Apparatus and equipment 2.5.1 High performance liquid chromatograph equipped with UV detector or DAD. 2. 5. 2 Shaker. 2.5.3 Nitrogen evaporator. 2. 5. 4 Rotary evaporator. 2. 5. 5 Vortex m

20、ixer. 2.5.6 Micro-syringe: 100L. 2. 6 Procedure 2. 6. 1 Extraction Weigh 25 9 (accurate to 0.01 g) of the test sample into a 250 mL conical flask with stopper, accu rately add 50.0 mL acetonitrile and set aside for 2 h. Filtrate after shaking for 30 min. Gather filtrate into a 100 mL measuring cylin

21、der in which 5 g. 7 9 sodium chloride are placed. Stopper the measur ing cylinder and shake vigorously for ca 1 min , then let stand for separating completely. Transfer 10.0 mL acetonitrile layer into a pear-shaped flask, condense it near to dryness with rotary evaporator in a bath of 50C ,dissolve

22、the residue with 10 mL acetone-n-hexane (10+90). 7 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 2. 6. 2 Clean up Condition the 5PE tube with 2 mL n-hexane , add the sample solution into the 5PE tube when the above solution arrive the sorbent surface, reject the effluents. Rinse the pear-shaped flask with 5 mL ac

23、etone + N-hexane (10 + 90) , reject the effluents. Then rinse the 5PE tube with 5 mL acetone + N hexane (20 + 80) and once again , collect the elution with 15 mL scale centrifugal tube. Condense near to drness with nitrogen evaporator in a bath of 50oC. The residue is dissolved with 2.0 mL acetoni t

24、rile + water (30 + 70) , and mixed with the vortex mix町，filteredthrough a 0.45m membrane and ready for HPLC determination. 2. 6. 3 Determination 2. 6. 3. 1 HPLC operating condition a) Column: OD5-C18 (5m) ,250 mm x 4 mm( i. d. ) or equivalent; b) Column temperture:40oC ; c) Mobile phase:acetonitrile

25、+ wat町，gradient:Omin ,5%; 15 min ,25% ;20 min,5%; d) Flow rate: 1.0 mL/min; e) Detector wavelength: 258 nm; f) Injection volume:20L 2. 6. 3. 2 HPLC determination According to the approximate concentration of the pesticide in the sample solution , select the mixed standard working solution with simil

26、ar peak area to that of the sample solution. The responses of pesticide in the mixed standard working solution and sample solution should be within the linear range of the instrumental detection. Under the above chromatographic condition, the retention time of imidacloprid and acetamiprid is ca 13 m

27、in and 15 min. For HPLC chromatogram of the standa时，seealso annex A. 2. 6. 4 81ank test The operation of blank test is the same as that described in the method of determination but with omission of sample addition. 2. 7 Calculation and expression of the result Calculate the content of imidacloprid o

28、r acetamiprid in the test sample according to the formula (门，8 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 the blank value should be substracted from the result of calculation. x=x c x V As x m . ( 1 ) where X -the residue content of imidacloprid or acetamiprid in the test sample , mg/kg; A-the peak area of imi

29、dacloprid or acetamiprid in the sample soluion; c-the concentration of imidacloprid or acetamiprid in the standard working solution，g/mL; V-the final volume of the sample solution , mL; As-the peak area of imidacloprid or acetamiprid in the standard working solution; m-the corresponding mass of the

30、test sample in the final sample solution ,g. 3 Li mit of determination and recove叩3. 1 Li mit of determination The limit of determination of this method is 0.02 mg/kg. 3. 2 Recovery 3. 2. 1 According to experimental data , the fortifing concentrations of imidacloprid and acetamiprid in tomato and th

31、eir corresponding recoveries are: 0.02 mg/kg ,the recovery of imidacloprid and acetamiprid is 91.4%-109% and 95.5%-108%; 0.05 mg/kg , the recovery of imidacloprid and acetamiprid is 89.2% -101 % and 90.0% -102%; 0.10 mg/kg ,the recovery of imidacloprid and acetamiprid is 80.7%-88.5% and 85.5%-103%.

32、3. 2.2 According to experimental data , the fortifying concentrations of imidacloprid and acetamiprid in cucumber and their corresponding recoveries are: 0.02 mg/kg ,the recovery of imidacloprid and acetamiprid is 98.5%-106% and 94.0%-106%; 0.05 mg/kg ,the recovery of imidacloprid and acetamiprid is

33、 95.0%-106% and 90.0%-102%; 0.10 mg/kg ,the recovery of imidacloprid and acetamiprid is 81.4%-97.2% and 96.2%-109%. 3. 2.3 According to experimental data , the fortifing concentrations of imidacloprid and acetamiprid in orange and thei r corresponding recoveries are: 0.02 mg/kg , the recovery of imi

34、dacloprid and acetamiprid is 94.5% -102% and 90.0% -99.0%; 0.05 mg/I甸，therecovery of imidacloprid and acetamiprid is 88. 0% -98.4% and 84.4% -93.8%; O. 10 mg/kg , the recovery of imidacloprid and acetamiprid is 84.9% -109% and 87.3% -99.5%. 9 版权所有 禁止翻制、电子传阅、发售SN/T 1902-2007 Annex A ( Informative) HP

35、LC chromatogram of the standards 古=巳【ESO革面。由自由飞【甘己且。-udu甘【口口坦白.NJmAU 0.2岳。0.25 一二)一一一一。.5o. 7岳-1 1.2岳一一-_-_. 1111n 17 nb l 15 14 13 12 11 10 9 8 1.岳Figure A. 1-HPLC chromatogram of the standards (0.08g/mL) 10 版权所有 禁止翻制、电子传阅、发售khCON-NO-问Z中华人民共和国出入境检验检疫行业标准水果蔬菜中毗虫琳、口比虫清残留量的测定高效液中目色谱法SN/T 1902-2007 -兴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当咛1/16 印张1字数17千字2007年9月第一版2007年9月第一次印刷印数1-2000 开本880X 1230 定价10.00元-兴书号:155066 2-18046 SN/T 1902-2007