基于葡萄糖/葡萄糖氧化酶/H_2O_2/L-酪氨酸/辣根过氧化物酶反应体系的荧光毛细分析法测定血糖

Determination of Blood Glucose by Fluorescence Capillary Analysis Based on Glucose/Glucose Oxidase/H_2O_2/L-tyrosine/ Horseradish Peroxidase Reaction System

建立了葡萄糖(G)/葡萄糖氧化酶(GOD)/H2O2/L-酪氨酸/辣根过氧化物酶(HRP)新荧光反应新体系,并将此反应体系与荧光毛细分析法(FCA)结合,开发一种能够真正实现血糖测定的新葡萄糖酶荧光毛细分析法(GE-FCA)。经优选得到的新反应体系最佳条件为:GOD和HRP浓度均为2000U/L,L-酪氨酸浓度为1.0mmol/L,磷酸盐缓冲液的pH=8.5,反应温度为35℃,反应时间为20min。通过测定葡萄糖标准样品,得到GE-FCA法测定葡萄糖的线性范围是5.0～140μmol/L;相对标准偏差<4.0%;检出限为2.65μmol/L。本方法已成功用于临床血清样品中葡萄糖的测定,其回收率在98%～104%之间。GE-FCA荧光增强法操作简单,样品和试剂用量极少(9μL),线性响应范围宽、抗干扰能力强,易于普及推广。

A novel fluorescent reaction system of glucose/glucose oxidase（GOD）/hydrogen peroxide/L-tyrosine/horseradish peroxidase（HRP） combined with a fluorescence capillary analysis（FCA） was established for developing a sensitive glucose enzyme FCA（GE-FCA）.After studying the new reaction,the optimum conditions were as follows： concentrations of GOD and HRP were both 2000 U/L,concentration of L-tyrosine was 1.0 mmol/L,pH of phosphate buffer solution was 8.5,reaction temperature was 35 ℃ and reaction time was 20 min.The GE-FCA′s linear range was 5.0-140 μmol/L,relative standard deviation was less than 4.0% and detection limit was 2.65 μmol/L.The method has been successfully applied for the determination of blood glucose and the recovery was in the range of 98%-104%.Compared with routine methods,merits of the GE-FCA was simple in operation,less in sample volume and dosage of chemical as well as enzymic reagents（only 9 μL）,wide in linear range,strong in anti-interference ability and low cost,it will be easily popularized.