科研之友
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摘要
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 degrees C; the acidity was pH 6.5; the concentration of NADH was 300 mu mol/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 2.0 mu L and 18.0 mu L, respectively, and its analytical speed is 30 samples per hour. The LDH's activity in the serums was successfully determined by utilizing this method.
- 出版日期2012-7
- 单位四川大学
