基质固相分散_高效液相色谱法测定水稻中痕量异丙威

1、 基金项目河北省教育厅资助项目作者简介杨容(1962-,女,教授,从事农药残留分析。【研究报告】基质固相分散-高效液相色谱法测定水稻中痕量异丙威杨容,马晶军,宋双居(河北农业大学理学院化学系,河北保定071001摘要研究出一种应用基质固相分散(MSPD -高效液相色谱法测定水稻中痕量异丙威残留物的新方法。并讨论了应用MSPD 技术进行前处理与传统残留分析的区别。本实验将MSPD 技术用于水稻中异丙威残留物的高效液色谱分析,并对定量检测条件作了详细的研究。关键词高效液相色谱;基质固相分散;异丙威;水稻中图分类号O65717+2文献标识码A 文章编号1004-8685(200501-0048-02

2、Matrix solid -phase dispersion (MSPDand high performance liquid chromatographic de 2termination of trace isoprocarb in riceYang Rong ,M a Jing 2jun ,Song S huang 2ju(Department of Basic Courses ,Agricultrure University Hebei ,Baoding 071001,China AbstractA simple and reliable method for the determin

3、ation of trace isoprocarb in rice by matrix solid -phase dispersion (MSPD and liquid chromatography was presented 1The MSPD -HPLC technique used for assay of trace isoprocarb in rice was discussed and the conditions of quantitative analysis investigated in detail 1K ey w ordsHPLC ;MSPD ;Isoprocarb ;

4、Residue ;Rice异丙威(isoprocarb 是氨基甲酸酯类杀虫剂,具有较强的触杀作用,常用于水稻害虫的防治1。近十几年来,食品中农药残留物在世界范围内引起了人们的关注,但是对异丙威残留物的研究较少,尤其是利用高效液相色谱法(HPLC 23。一般应用HPLC 对食物中某种痕量物质进行测定时,样品的前处理都比较复杂。美国Austin RL 等4报道的基质固相分散(MSPD 前处理提取分离技术,样品吸附到很大面积上(500m 2/g C 18,然后依靠所选定的溶剂依次洗脱,可大大简化许多残留物前处理方式,最大限度地避免被测残留物的流失。本试验将MSPD 技术用于水稻中异丙威残留物的HPL

5、C 分析,并对定量检测条件作了详细的研究。1试验材料与方法111仪器及试剂高效液相色谱仪由岛津(Shimadzu LC -6A 高压输液泵,岛津SPD -6AV 紫外可见分光检测器,7125注射进样阀,PEHS -5C 18(5m ,416mm ×125mm 色谱柱,台式自动平衡记录仪组成。100140目ODS 按文献5,6方法合成,产品经碳元素分析测定符合要求。环己烷、丙酮、甲苯、甲醇均为分析纯试剂,使用前经过蒸馏提纯。甲醇经0145m 滤膜过滤。水经3次蒸馏。异丙威标样由农业部农药检测所提供。112色谱条件流动相:甲醇/水(v/v ,65/35;流量:110ml/min ;记录纸

6、速:4mm/min ;进样体积:10l ;紫外检测波长:254nm 。113基质柱的制备取1g C 18键合相与0125g 样品及异丙威标准溶液(25l 质量浓度为300mg/L 异丙威混合均匀,得到ODS/大米粉混合物,称其为基质。取1支2ml 注射器中,于注射器底部垫一圆滤纸片,将基质填入后在注射器顶部再加一圆滤纸片以防止基质流出。然后用注射器塞加压至最后体积为115ml ,制得基质柱。2结果与讨论211洗脱剂用量的选择首先用810ml 分析纯环已烷淋洗按前述方法制备的基质柱。在所有情况下洗脱液的流动是重力控制。如果洗脱液流动不畅,则在柱顶用吸液球加压,以推动重力流动。当洗脱液流出停止后,

7、柱内剩余的环已烷用上述加压法从柱上除去。然后用8ml 甲苯洗脱异丙威,收集洗脱液,每1ml 1份,共收集8份,分别测定各份中异丙威的含量(以峰高表示。实验结果表明,6ml 甲苯就可将异丙威提取完全。用8ml 环已烷清洗基质柱,可除去大米中的油质、蜡质及色素。对清洗液作高效液相色谱分析,结果表明环已烷中无异丙威存在。然后用6ml 甲苯洗脱,洗脱液用无油压缩空气流缓缓吹干。这一步骤很重要,否则在进行测定时会出现杂峰。吹干后的农药残留物用015ml 甲醇溶解,1700r/min 离心10min ,取上清液(10l 进行液相色谱分析。212流动相的确定实验表明,以甲醇-水为流动相洗脱效果好。进一步检验

8、了不同配比的甲醇/水二元流动相的洗脱效果,结果以体积比为65/35(v/v 的甲醇/水作流动相为宜。213检测波长的选择取适当浓度的异丙威的标准溶液在200350nm 范围内(下转第123页的效率。蛋白质组学的出现虽然只有几年时间,但已逐步形成产业和市场。2003年生命科学产业著名市场研究公司美国加州前线战略管理咨询公司预测2008年全球蛋白质组市场将达到2618亿美元。未来几年,蛋白质组市场综合年增长率将达到12%。总之,蛋白质组分析技术将越来越多地应用于医学研究中,为探索生命的奥秘和疾病的发生机理做出贡献。参考文献1OFarrell PH1High resolution two-dimen

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17、d proteinsin human nasal lavage fluid from non-smokers and smokers using two-dimensional gel electrophoresis and peptide mass flngerprinting J.Proteomics,2002,2:112-1201(收稿日期:2004-06-02(上接第48页进行紫外吸光度扫描,观察到异丙威的最大吸收峰分别在208nm和254nm处,由于208nm接近甲醇的紫外光截止波长,故选择254nm作为检测波长。214工作曲线、检出限和回收率用甲醇作溶剂,准确配制一系列异丙威的标准

18、溶液,按选定的条件实验。当异丙威的进样量为30900ng时,以异丙威峰高(cm对其进样量X(ng进行线性回归,求得线性回归方程为Y=01315+01950X,相关系数为01995;检出限(以2倍信噪比计算为0186ng。于一定体积的提取液中添加适量的标准品后进行回收率试验,结果见表1。表1异丙威的回收率农药添加浓度(ng/g回收率(%(x±s,n=5平均回收率(%异丙威15030045060075090088107±413089100±510088125±412087109±511488150±417588135±411088121±4158215样品测定对不同地区的大米样品进行测定,每个样品测定6次,样品中异丙威的残留量分别为:天津小站米1190ng/g,浙江亿禾大米2180ng/g,安徽梗稻米0180ng/g,黑龙江七河源大米未检出。3讨论上述试验表明,应用