荧光素二磷酸酯的合成和荧光分光光度法分析碱性、酸性磷酸酯酶

Synthesis of Fluorescein Diphosphate and Its Analysis of Alkaline and Acidic Phosphatase by Fluorimetry

目前荧光光度法测定碱性和酸性磷酸酯酶的底物常用伞形酮磷酸酯和萘酚As-BI(6-溴-3-羟基-2-萘基邻茴香胺)的磷酸二氢酯.它们具有很高的灵敏度,在荧光免疫分析中被广泛应用. 本文用荧光素和三氯氧磷在吡啶介质中合成了荧光素二磷酸酯,再用氢氧化钠处理为四钠盐.该醇易溶于水,其水溶液无荧光.它在碱性或酸性磷酸酯酶的催化下水解,产生具有很强荧光的荧光素,且荧光强度与酶量成正比,因此可进行碱性或酸性磷酸酯酶的测定.

Fluorescein diphosphate was synthesized with fluorescein and phosphorus oxychloride. The diphosphate was applied to the fluorimetric analysis of alkaline and acidic phosphatase. The conditions of reaction of phosphatase with fluorescein diphosphate, linear range for the determination of the phosphatase, limitation of detection of the phosphatase and Michaelis constant were studied.