酶荧光毛细管分析法测定微量血清中乳酸脱氢酶活性

基于丙酮酸/还原型辅酶I/乳酸脱氢酶（LDH）/乳酸/氧化型辅酶I荧光猝灭体系和荧光毛细管分析技术,建立了可用于微量样品中LDH酶活性测定的方法。优化的测定条件为：激发及发射波长分别为350和460nm;测定温度为25℃;酸度为pH 6.5;NADH浓度为300μmol/L;丙酮酸浓度为1.2mmol/L。本方法的测定范围为50～1500IU/L,检出限为30IU/L,相对标准偏差2.1%～2.2%（n=10）,回收率在96.4%～105%范围内。本方法操作简单,每次测定仅需样品2.0μL、试剂18.0μL,分析速度约为30样/h,利用本方法测定了微量血清中LDH的活性。...

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Bibliographic Details
Published in分析化学 Vol. 40; no. 7; pp. 1043 - 1047
Main Author 李乔婧 李永生 周朗 杨微 高秀峰
Format Journal Article
LanguageChinese
Published 四川大学化学工程学院,成都,610065%四川大学华西基础医学与法医学院,成都,610041 2012
Subjects
乳酸脱氢酶
荧光毛细管分析
血清
酶活性
乳酸脱氢酶
荧光毛细管分析
血清
酶活性
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ISSN0253-3820
DOI10.3724/SP.J.1096.2012.11154

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Summary:基于丙酮酸/还原型辅酶I/乳酸脱氢酶（LDH）/乳酸/氧化型辅酶I荧光猝灭体系和荧光毛细管分析技术,建立了可用于微量样品中LDH酶活性测定的方法。优化的测定条件为：激发及发射波长分别为350和460nm;测定温度为25℃;酸度为pH 6.5;NADH浓度为300μmol/L;丙酮酸浓度为1.2mmol/L。本方法的测定范围为50～1500IU/L,检出限为30IU/L,相对标准偏差2.1%～2.2%（n=10）,回收率在96.4%～105%范围内。本方法操作简单,每次测定仅需样品2.0μL、试剂18.0μL,分析速度约为30样/h,利用本方法测定了微量血清中LDH的活性。
Bibliography:Enzyme activity, Lactate dehydrogenase, Fluorescence capillary analysis, Serum
LI Qiao-Jing , LI Yong-Sheng , ZHOU Lang , YANG Wei , GAO Xiu-Feng 1 （School of Chemical Engineering, Sichuan University, Chengdu 610065, China） 2 （West China School of Preclinical and Forensic Medicine, Sichuan University Chengdu 610641, China）
22-1125/O6
Based on a fluorescence quenching reaction system of pyruvic acid/reduced form of nicotinamide-adenine dinucleotide （NADH）/lactate dehydrogenase （LDH）/lactic acid/oxidized form of nicotinamide-adenine dinucleotide and a technology of the fluorescence capillary analysis, we proposed a new method for determining the LDH＇s activity in micro-volume samples. First, the optimum conditions for the method were as follows： the excitation and the emission wavelength were 350 nm and 460 nm respectively; the temperature was 25 ℃; the acidity was pH 6.5; the concentration of NADH was 300 gmol/L; the concentration of pyruvate was 1. 2 mmol/L. Afterwards, the measurement range （50--1500 IU/L）, the
ISSN:0253-3820
DOI:10.3724/SP.J.1096.2012.11154