表面活性剂对NBD-Cl衍生氨基酸的荧光增敏和芯片电泳分离的影响  

Effect of Surfactants on Detection and Separation of 4-Cl-7-nitrobenzofurazan Derived Amino-acids by Chip-based Capillary Electrophoresis

作　　者：李志明[1] 何巧红[1] 陈恒武[1] 

机构地区：[1]浙江大学化学系微分析系统研究所,杭州310058

出　　处：《分析化学》2008年第7期967-970,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No20475048);浙江省自然科学基金(NoY404316)资助项目

摘　　要：研究多种表面活性剂以及溶剂对4-氯-7-硝基苯-2-氧-1,3-二唑(NBD-Cl)衍生氨基酸荧光性质的影响。考察并优化了NBD-Cl与氨基酸的衍生反应条件以及芯片毛细管电泳-激光诱导荧光的分离分析条件。结果表明:在临界胶束浓度以上,阳离子表面活性剂十六烷基三甲基溴化胺(CTMAB)对精氨酸、谷氨酸、天冬氨酸的NBD-Cl衍生物,非离子表面活性剂十二烷基聚氧乙烯醚(Brij-35)对苯丙氨酸、精氨酸的NBD-Cl衍生物有荧光增敏效果。在含有5mmol/LBrij-35和20%乙腈添加剂的15mmol/L硼砂缓冲液(pH9.2)中,精氨酸、苯丙氨酸、丙氨酸、甘氨酸、谷氨酸及天冬氨酸等6种氨基酸的NBD-Cl衍生物可以在70s内实现分离,其中较难分离的丙氨酸、甘氨酸的NBD-Cl衍生物分离度为0.91;6种氨基酸衍生物的检出限为0.15～1.6μmol/L。The effect of various surfactants and solvents on the fluorescence properties of the 4-chloro-7-nitrobenzofurazan（NBD-Cl） derived amino-acids was studied.The experimental conditions for the derivatization reaction between NBD-Cl and amino acids,and those for chip-based capillary electrophoresis separation-laser induced fluorescence detection（CCE-LIF） of the amino acid derivatives were optimized.It was found that above their critical micelle concentrations,cationic surfactant CTMAB sensitized the fluorescence intensity of NBD-Cl derived arginine,glutamic acid,and aspartic acid,while non-ionic surfactant Brij-35 did the fluorescence intensity of the NBD-Cl derived phenylalanine and arginine.It was observed that six NBD-Cl derived amino acids of arginine,phenylalanine,alanine,glycine,glutamic acid and aspartic acid were CCE separated within 70 s in a 15-mmol/L pH 9.2 borate buffer containing 5 mmol/L Brij-35 and 20% acetonitrile additives,the resolution of the partially overlapped alanine and glycine derivatives being 0.91,and the detection limits of the six derived amino acids were 0.27-1.6 μmol/L.

关 键 词：氨基酸 4-氯-7-硝基苯-2-氧-1 3-二唑 芯片毛细管电泳 激光诱导荧光 表面活性剂 荧光增敏 

分 类 号：Q517[生物学—生物化学]