毛细管电泳-激光诱导荧光检测法分离检测谷胱甘肽及其构成氨基酸  

Separation and detection of Glutathione,Glutamic acid,Cysteine,and Glycine with laser-induced fluorescence detection in capillary electrophoresis

作　　者：王宇飞[1] 蔡元丽[1] 林夏[1] 晏瑾[1] 李晖[1] 

机构地区：[1]四川大学化学工程学院,四川成都610065

出　　处：《化学研究与应用》2013年第3期311-316,共6页Chemical Research and Application

摘　　要：以4-氯-7-硝基苯并-2-氧杂-1,3-二唑(NBD-Cl)为柱前衍生试剂,建立了一种毛细管电泳-激光诱导荧光直接检测氧化型和还原型谷胱甘肽及其构成氨基酸(谷氨酸、半胱氨酸和甘氨酸)的新方法。经过实验条件的优化,采用25 mmol/L硼砂-20 mmol/L聚氧乙烯月桂醚(Brij-35)-5%乙腈(pH 9.5)的缓冲体系,在柱温为25°C、分离电压为20 kV的条件下,压力进样3447.5 Pa(0.5 psi)×3 s,五种物质在11 min内实现高效基线分离。在该方法下,还原型谷胱甘肽、氧化型谷胱甘肽、谷氨酸、半胱氨酸和甘氨酸的线性范围分别为:1～50μg/mL,1～50μg/mL,0.5～25μg/mL,1～50μg/mL,0.1～20μg/mL;检测限分别为:0.1μg/mL,0.1μg/mL,0.005μg/mL,0.1μg/mL,0.001μg/mL。以还原型谷胱甘肽钠粉针剂为样品,方法的加标回收率为99.5%～110.7%,相对标准偏差为0.26%～3.272%(n=3)。该方法准确、快速、灵敏、检测限低,有望用于样品中氧化型和还原型谷胱甘肽及其构成氨基酸的含量分析。A capillary electrophoresis method with laser-induced fluorescence detection（LIFD） was developed for the determination of oxidation and reduced glutathione, glutamic acid, cysteine and glycine after derivatization with 4-chloro-7-nitrebenz0furazan （NBD-C1）. The five chemicals could be well separated with in 11 min at a voltage of 20 kV with 25 ~C cartridge temperature in a running buffer consisting of 25 mmoL/L berate sodium ,20 mmoL/L Brij-35 and 5% acetonitrile（ pH9.5 ） ,and the samples were in- jectod by pressure 3447.5 Pa（0.5 psi） x 3 s. Under the optimum condition,the method has good linearity relationships（ square of correlation coefl＇icients ：0. 9994-0. 9999 ） with ranges of 1-50 p^g/mL for GSH, GSSG, Cys; 0. 5 ~ 25 p.g/mL for Glu and 0. 1 ~ 20 p^7＇mL for Gly, respectively. The limit of detection（LOD） for ana]ytes were in the range of 0. 001-0. 1 p^g/mL This method was ap- plied to the quantitation of GSH powder injection with the recoveries in the range of 99. 5% ~ 110. 7% and the relative standard de- viation（RSD） were 0. 26% -3. 272% （n = 3 ）. This method is accurate ,fast, sensitive and has a low detection limit. It can be used for the content of oxidation and reduced glutathione and amino acids that form the glutathione in samples. This method has a good prospect in the analysis of real samples.

关 键 词：胶束电动色谱-激光诱导荧光检测法 多肽 氨基酸 4-氯-7-硝基苯并-2-氧杂-1 3-二唑(NBD-Cl) 

分 类 号：O657.7[理学—分析化学]