1、中华人民共和国出入境检验检疫行业标准SN/T 1866-2007 进出口粮谷中咪瞠磺隆残留量检测方法液相色谱法Determination of imazosulfuron residue in cereals for import and export-Liquid chromatography 2007-04-06发布2007-10-16实施中华人民共和国发布国家质量监督检验检度总局目IJ1=1 本标准的附录A为资料性附录。本标准由中华人民共和国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天津出入境检验检疫局负责起草。本标准主要起草人:张曼、古珑、何佳、张骏、许154、林安清
2、、吴延晖。本标准系首次发布的检验检疫行业标准。SN/T 1866-2007 1 范围进出口粮谷中咪瞌磺隆残留量检测方法液相色谱法本标准规定了出口粮谷中咪嗖磺隆残留量检验的制样和液相色谱检测方法。本标准适用于出口糙米中眯嗖磺隆残留量的检测。2 测定方法2.1 方法提要SN/T 1866-2007 用二氯甲:皖、乙睛和氨水的?昆合溶剂提取试样中残留的咪唾磺隆，提取液经Cs固相萃取柱净化，液相色谱紫外检测器检测，外标法定量。2.2 试剂和材料除另有规定外，所用试剂均为分析纯，水为二次蒸馆水。2.2.1 甲醇:高效液相色谱级。2.2.2 乙睛:高效液相色谱级。2.2.3 二氯甲:院。2.2.4 冰乙酸
3、。2.2.5 1 mol/L氨水:将70mL浓氨水用水稀释至1L2.2. 6 C18固相萃取柱:3mL, 200 mg填料。使用前用3mL甲醇和3mL水依次预淋洗。2.2.7 提取液:二氯甲:皖+乙腊+1mol/L氨水(4十15+1，体积比)。2.2.8 t先脱液:甲醇十水(1十1，体积比)。2.2.9 咪嗖磺隆标准品(imazosulfuron，C14 Hll CINc 0 S, CA: 122548-33-8) :纯度大于等于99%。2.2. 10 咪瞠磺隆标准溶液:准确称取适量的咪唾磺隆标准品，用乙腊配成浓度为100g/mL的标准储备液。根据需要用流动相稀释成适当浓度的标准工作液。2.3
4、 仪器和设备2.3. 1 液相色谱仪配有紫外检测器。2.3.2 振荡器。2.3.3 旋转蒸发器。2.3.4 氮吹仪。2.3.5 固相萃取装置。2.4 测定步骤2.4. 1 制样将样品按四分法缩分至1kg，全部磨碎并过筛，筛孔0.90mm(20目)，混匀，均分成两份同样，装入洁净的容器内，密封，标明标记。于50C以下避光保存。在制样过程中，应防止样品受到污染或发生咪瞠磺隆残留量的变化。2.4.2 提取称取试样10g(精确到O.1 g)于250mL具塞锥形瓶中，加入50mL提取液(2.2. 7) ，在振荡器上提取30mi口，过滤至250mL鸡心瓶中。样品残渣重复上述操作一次。再用10mL二氯甲皖洗
5、涤滤纸，收集滤液于同一鸡心瓶中。在400C水浴中用旋转蒸发浓缩至干。用4mL乙腊溶解残渣，将溶解液转移SN/T 1866-2007 至10mL刻度试管中。在氮吹仪上于40C通氮气浓缩至干，用0.2mL甲醇重新溶解，再用水稀释至10 mL。2.4.3 净化将稀释液全部通过小柱，用5mL水淋洗，淋洗后用12mL洗脱液(2.2.8)洗脱，收集全部洗脱液于100 mL鸡心瓶中，在40C水浴中旋转蒸发至干。用1.0 mL乙睛溶解残渣，经O.45m滤膜过滤后供液相色谱测定。2.4.4 测定2.4.4. 1 液中目色谱条件a) 色谱柱:Cj日，150mmX4. 6 mm , 3. 5m或相当者;b) 流动相
6、:乙腊+0.1%乙酸(45十日，体积比); c) 流速:1.0 mL/min; d) 柱温:25C;e) 检测波长:238口m;f) 进样量:20L。2.4.4.2 液中目色谱测定根据样液中咪唾磺隆含量情况，选定峰面积相近的标准工作溶液。标准工作溶液和样液中咪唾磺隆响应值均应在仪器检测线性范围内，标准工作、溶液和样液等体积参插进样测定。在上述色谱条件下，咪嗖磺隆的保留时间约为6.6mi口。标准品的液相色谱图参见附录A中图A.1 2.4.5 空白试验除不加试样外，均按上述步骤进行。2.4.6 结果计算和表述用色谱数据处理机或按式(1)计算试样中咪瞠磺隆残留量，计算结果应将空白值扣除。V一-m f
7、hu一-A A一X . ( 1 ) 式中:X 试样中咪瞠磺隆的残留量，单位为毫克每千克(mg/kg); A 样液中眯哩磺隆的峰面积，单位为平方毫米(mm2); A 标准工作溶液中咪嗖磺隆的峰面积，单位为平方毫米(mm2); Cs 标准工作榕液中咪嗖磺隆的浓度，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);m 最终样液所代表的试样量，单位为克(g)3 方法的测定低限、回收率3.1 测定低限本方法的测定低限为0.02mg/kg o 3.2 回收率糙米中眯哩磺隆的添加浓度及回收率的实验数据:咪瞠磺隆的添加浓度在O.02 mg/kgO. 50 mg/kg时，回收率范围为88
8、.5%90. 7%。2 0.005 0.010 附录A(资料性附录)标准品色谱固AU 0.040 SN/T 1866-2007 0.03岳0.030 0.025 0.020 0.015 nu l nu nu 0.00岳EE由自由lAHliti-、ii也铿营运nu nu nu nu t/min 1. 00 2. 00 3. 00 4. 00岳006.00 7. 00 8.00 9.00 10.00 图A.1咪瞌横隆标准晶液相色谱图3 SN/T 1866-2007 Foreword Annex A of this standard is an informative annex. This sta
9、ndard was proposed by and is under the charge of China National Regulatory Commission for certification and Accreditalion. This standard was drafted by Tianjin Entry-Exit inspection and Quarantine Bureau of the peoples Re public of China. The main drafters of this standard are Zhang man ,Gu long , H
10、e jia , Zhang jun ,Xu hong ,Lin anqing. This standard is a professional standard promulgated for the first time. 4 SN/T 1866-2007 Determination of imazosulfuron residue in cereals for import and export-Liquid chromatography 1 Scope This standard specifies the methods of sample preparation and determ
11、ination bliquid chromatogra phy of imazosulfuron residue in cereals for import and export. This standard is applicable to the determination of imazosulfuron residue in rice for import and export. 2 Method of determination 2. 1 Principle The imazosulfuron residue were extracted with dichloromethane,
12、acetonitrile and ammonia , purified by C18 SPE column , determinated by HPLC with UV-detector, Calculated by comparing peak of the sample with corresponding standard peak area. 2. 2 Reagents and materials Unless otherwise specified ,all reagents used should be analtically pure. Water is redistilled
13、Water. 2.2. 1 Methanol: HPLC grade. 2.2.2 Acetonitrile: HPLC grade. 2.2.3 Diloromethane. 2.2.4 Acetic acid. 2.2.5 1 mol/L Ammonia: Pipet 70 mL ammonia to volumetric flask and dilute to 1 L. 2.2.6 C18 SPE column: 3 mL/200 mg , condition the SPE cartridge with 3 mL methanol followed by 3 mL water. 2.2
14、.7 Extract solvent: Dichloromethane + Acetonitrile + 1 mol/L Ammonia(4 + 15 + 1 , V / V / V). 2.2.8 Eluant: Methanol + waterC 1 + 1 , V / V). 5 SN/T 1866-2007 2.2.9 Imazosulfuron standard (C11 H13CIN605S,CA: 122548-33-8): Purity二三99%.2.2.10 Imazosulfuron standard solution: Weigh appropriate imazosul
15、furon , dissolve in acetonitrile and prepare a solution of 100g/mL as standard stock solution. Dilute the imazosulfuron standard stock solution to the required concentration as the standard working solution with mobile phase. 2.3 Apparatus and equipment 2.3.1 HPLC: Equipped with UV-detector. 2.3.2 O
16、rbital shaker. 2.3.3 Rotation evaporator. 2.3.4 Nitrogen evaporator with heated bath. 2.3.5 Solid phase extraction equiment. 2.4 Procedure 2.4. 1 Sample preparation Reduce the sample to ca 1 kg by quartering , grind thoroughly and let pass through a 20-mesh sieve, mix thoroughly and divide into 2 eq
17、ual portion , place in clean containers , seal and labe l. The test sample shall be stored below - 5C and kept away from light. In the course of sample prepa ration , precaution must be taken to aviod contamination or any factors which may cause the change of residue content. 2.4. 2 Extraction Weigh
18、 10 9 (accurated to O. 1 g) of the test sample into 250 mL taper bottle, adding 50 mL extract solvent (2.2.7). It was coverd and shaked for 30 min. The extraction liquid was filterated to 250 mL erlenmeyer flake. Repeated above operation. Then , the filter paper was washed b10 mL dlorometh ane to th
19、e same erlenmeyer flake. Rotary evaporate the extraction liquid at 40C to dry, dissolve the residues with 4 mL acetonitrile, tranfer to 10 mL graduated tube ,dry under nitrogen at 40C. Redis solve the residues with 0.2 mL methanol ,distilled to 10 mL with water. 2.4.3 Clean up Transfer above solutio
20、n to the cartridge. Wash cartridge with 5 mL water. Then transfer 12 mL eluant (2.2.8) to cartridge , rotary evaporate the elution liquid at 40C to dr. Dissolve the residues with 6 SN/T 1866-2007 1.0 mL acetonitrile. It was filtrated with 0.45m filter, finallwas subjected to analsis. 2.4.4 Determina
21、tion 2.4.4. 1 LC operating conditions a) Column:CB 150 mm x 4.6 mm(i. d. ) ,3. 5m or equivalent; b) Mobile phase:Acetonitrile一0.1%Aceticacid (45+55 , V/V); c) flowrate:1.0mL/min; d) Column temperature:250C ; e) Wave length of UV detecter: 238 nm; f) Injection volumn: 20L. 2.4.4. 2 LC determination A
22、ccording to the approximate concentration of imazosulfuron residuces in sample solution , select the standard working solution with similar peak area to that of the sample solution. The standard working solution should be randomly injected in between the injection of sample solution of equal volumn.
23、 Un der the above LC conditions , the retention time of imazosulfuron is about 6.6 min. See Figure A. 1 in annex A. 2. 4. 5 81ank test Perfore the blank test with the same procederes as that described in the method of determination but without addition of test sample. 2.4. 6 Calculation and expressi
24、on of the results The calculation of results is accoring to the formula( 1) : V一-m A一-A A一X 、，Fall f . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Where , X -the residue of imazosulfuron , mg/kg; A -the peak area of imazosulfuronof the sample solution , mm勺As-the peak area of imazosulfur
25、on the Standard working solution，mm勺cs-the concentration of imazosulfuron in the Standard working solution，g/mL; 斤1-the门lassof test sa门lplein the final solution , g. 7 SN/T 1866-2007 3 Li mit of determination and recovery 3. 1 limit of determination Li mit of determination is 0.02 mg/kg. 3. 2 Recove
26、ry According to the experimental data , the fortifing concentration of imazosulfuorn at 0.02 mg/kg-0.50 mg/闸，andits corresponding recoveries are 88.5% -90.7%. 8 AU 0.040 Annex A Cinformative annex) Chromatogram of standard 0.03岳才口E z E 0.030 才E i ilA 由咀0.020 0.015 0.010 0.00岳0.000 0.005 0.010 SN/T 1
27、866-2007 1. 00 2.00 3.00 4.00岳006.00 7. 00 8.00 9.00 10.00 t/min Figure A. 1-The HPLC chromatogram of imazosulfuron standard 9 khCON-问Z中华人民共和国出入境检验检疫行业标准进出口粮谷中咪瞌磺隆残留量检测方法液相色谱法SN/T 1866-2007 -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7月第一版2007年7月第一次印刷印数1-2000 开本880X 1230 定价10.00元-兴书号:155066.2-17831 SN/T 1866-2007
