荧光毛细分析法测定丙酮酸的研究

Research on determination of pyruvic-acid using fluorescence capillary analysis

基于荧光毛细分析法（Fluorescence Capillary Analysis，FCA）和液态酶（Hqllid enzylne method，LE）催化反应，开发了一种荧光毛细法测定丙酮酸的方法（P-LE-FCA）。在激发波长350nm、发射波长460nm，磷酸缓冲液pH=7．50，温度38℃，酶浓度5．0kUL^-1，NADH为1．0mmol·L^-1的条件下，成功对尿样中的丙醌酸进行了测定。标准曲线的线性范围为0．2mmol·L^-1～1．2mmol·L^-1，r=0．9942，检出限为0．012mmol·L^-1。P-LE-FCA法操作简便，样品用量仅为18μL，测定成本低，可实现丙酮酸的微量化测定。有望将酶固定在毛细管壁上，制成荧光毛细丙酮酸测试盒，用于含丙酮酸的实际样品的测定。

A new method （ P - LE - FCA） for the determination of pyruvic - acid has been proposed based on liquid enzyme method （LE） and Fluorescence Capillary Analysis （FCA）. Pyruvic - acid concentration was determined at 350 nm of excited wavelength , 460 nm of emission wavelength. The experimental conditions are as follows. The reaction time is 20 min; temperature is 38℃. The pH of phosphate buffer solution is 7. 50 and NADH concentration is 1.0 mmol·L^-1. The concentration of LDH is 5.0 kU·L^-1. The linear range is 0. 2mmol·L^-1 - 1.2 mmol· L^-1 （r =0. 9942） ,and its detection limit is 0. 012 mmol· L^-1. The sample volume is only 18μL. P - LE - FCA has the advantages such as lower cost, simple to operation, and it can realize micro and rapid determination. It can be made of a test kit for detecting the samples containing pyruvic - acid.