高通量蛋白质沉淀技术的开发及其在药物分析中的应用  

Development of high throughput protein precipitation technology and its applications in pharmaceutical analysis

作　　者：包建民[1] 张梦娜[1] 马志爽[1] 李优鑫[1] 

机构地区：[1]天津市现代药物传递及功能高效化重点实验室,天津化学化工协同创新中心,天津大学药物科学与技术学院,天津300072

出　　处：《分析试验室》2014年第10期1117-1120,共4页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金资助项目(21375093);高等学校博士学科点专项科研基金资助项目(20130032120081);天津大学自主创新基金资助项目(1102213)资助

摘　　要：针对传统离心沉淀蛋白质方法,建立了一种新的生物样品预处理方法 -高通量蛋白质沉淀技术（PPT）。以特殊的膜作为分离介质,以96孔板为支撑,结合高通量过滤技术,根据有机溶剂沉淀原理,快速高通量祛除生物样品中蛋白质。通过与传统蛋白质沉淀-离心法的对比,验证了该技术的可行性和稳定性,并进一步将该技术应用于实际血清样品中头孢氨苄和马来酸氯苯那敏两种药物浓度的检测。经该方法处理后头孢氨苄和马来酸氯苯那敏的回收率在80.9%~82.4%和96.8%~110.4%之间,样品间RSD值在0.91%~6.8%和1.1%~1.8%之间。A novel high-throughput protein precipitation technique （PPT） was established. It took a special membrane as separation medium and achieved a high throughput sample preparation method combined with highthroughput filtration technology and 96-well plates under the appropriate conditions of precipitator and serum concentration. The principle of the technology is organic solvent precipitation. The accuracy and feasibility of the PPT method were verified through the comparison with the traditional centrifugation method. This method is evaluated through rapid treatments of cephalexin and maleate chlorpheniramine in serum combined with HPLC quantification analysis. The results showed that the new method exhibited good spike recoveries range of 80. 9% to 82.4 % and 96. 8% to 110. 5%, with RSD ranges from 0. 91% to 6. 8% and 1.1% tol. 8 %. These results demonstrate that the novel protein precipitation technique is a rapid, simple, robust, easily automated and high- throughput technique for sample pretrea associated with traditional centrifugation tment alleviating many of the liquid handling and time consuming issues method.

关 键 词：样品预处理 生物样品 高通量 蛋白质沉淀 有机沉淀 

分 类 号：O652.6[理学—分析化学]