高等学校化学学报

高等学校化学学报 ›› 2001, Vol. 22 ›› Issue (4): 547.

• 研究论文 • 上一篇    下一篇

微量乳酸脱氢酶毛细管电泳在线反应电化学检测方法的研究

杨文初, 俞爱民, 陈洪渊   

  1. 南京大学化学系分析科学研究所, 南京 210093
  • 收稿日期:2000-02-21 出版日期:2001-04-24 发布日期:2001-04-24
  • 通讯作者: 陈洪渊(1937年出生),男,教授,博士生导师,从事电分析化学研究.
  • 基金资助:

    国家自然科学基金(批准号:29835110)资助.

Studies on an On-line Reaction of Micro Lactate Dehydrogenase in Capillary Electrophoresis Using Electrochemical Detection

YANG Wen-Chu, YU Ai-Min, CHEN Hong-Yuan   

  1. Department of Chemistry, Institute of Analytical Science, Nanjing University, Nanjing 210093, China
  • Received:2000-02-21 Online:2001-04-24 Published:2001-04-24

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被引次数

22

摘要： 以乳酸脱氢酶催化乳酸与NAD+反应生成丙酮酸与NADH和安培法检测NADH为基础,利用电泳中介微分析(EMMA)技术,研究了超微量乳酸脱氢酶的毛细管电泳在线反应的电化学检测方法,并从理论上对EMMA电泳图中的平台宽度和高度作了初步探讨.结果表明,在EMMA的恒高压和零高压两种模式下,使用直径为150μm和束状碳纤维电极,在+0.8V检测电位下,对LDH活性检测灵敏度分别为1.1nU和0.6nU;所导出的平台高度、宽度与实验条件的关系式对提高毛细管电泳分离效率和检测灵敏度有一定指导意义.

关键词: 毛细管电泳, 电化学检测, 乳酸脱氢酶

Abstract: The method for an on-line reaction of micro lactate dehydrogenase in capillary electrophoresis using electrophoretically mediated micro-assay(EMMA) coupled with electrochemical detection(EC) was developed. The running electrolyte(pH 9.2) was composed of 50 mmol/L Tris HCl, 10 mmol/Lnicotinamide adenine dinuceotide(NAD+), 5.0 mmol/Llithium lactate and 0.2 mmol/Lcetylpridinium bromide(CPB). LDHwas injected into the capillary. As it migrated electrophoretically in the capillary, LDH catalyzed the reaction between lactate and NAD+. The product NADH of the enzyme-catalyzed reaction was monitored by a bundle of carbon fiber electrode(150 μm diameter) held at +0.8 Vvs. SCE. The detection limits for LDHactivity using constant voltage mode and zero-voltage mode in EMMAwere determined to be 11×10-9 Uand 60×10-10 U, respectively. The formulae concerning the width or the height of the plateau in the electropherogram for EMMAwere deduced, and some significant conclusions were drawn from these formulae. The developed method has been used for the determination of ultramicro LDHactivity in serum with satisfactory results.

Key words: Capillary electrophoresis, Electrochemical detection, Lactate dehydrogenase

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