分析化学2012，Vol.40Issue(7)：1043-1047,5.DOI:10.3724/SP.J.1096.2012.11154
酶荧光毛细管分析法测定微量血清中乳酸脱氢酶活性
An Novel Enzyme Fluorescence Capillary Analysis for Determination of Lactate Dehydrogenase Activity in Micro-volume Serums
李乔婧 1李永生 1周朗 1杨微 1高秀峰2
作者信息
- 1. 四川大学化学工程学院,成都610065
- 2. 四川大学华西基础医学与法医学院,成都610041
- 折叠
摘要
Abstract
Based on a fluorescence quenching reaction system of pyruvic acid/reduced form of nicotinamide-adenine dinucleotide (NADH)/laetate 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 μmol/L; the concentration of pyruvate was 1. 2 mmol/L. Afterwards, the measurement range (50-1500 IU/L), the detection limit (30 IU/L), the relative standard deviation (2.1%- 2. 2%, n=10) and the recovery (96. 4% -105%) of the method were attained. This method is easy to operate, the dosages of the sample and reagents are only 2. 0μL and 18.0 μL, respectively, and its analytical speed is 30 samples per hour. The LDH's activity in the serums was successfully determined by utilizing this method.关键词
酶活性/乳酸脱氢酶/荧光毛细管分析/血清Key words
Enzyme activity, Lactate dehydrogenase, Fluorescence capillary analysis, Serum引用本文复制引用
李乔婧,李永生,周朗,杨微,高秀峰..酶荧光毛细管分析法测定微量血清中乳酸脱氢酶活性[J].分析化学,2012,40(7):1043-1047,5.
