荧光光度法测定血清中碱性磷酸酶

Spectrofluorimetric Determination of Alkaline Phosphatase in Human Serum

合成了一种新的底物水杨酸磷酸酯(SP),用于荧光光度法测定血清中碱性磷酸酶(ALP)活力。在37.0℃的Tris-HCl缓冲液(pH9.0)条件下,碱性磷酸酶作用于荧光底物SP,水解生成强荧光产物水杨酸(SA),生成的SA的量与参与反应的ALP的活力成正比。据此建立了荧光光度法测定血清中碱性磷酸酶活力的新方法。测定碱性磷酸酶的线性范围是0.03～6.00 U/L,检测限为7.04 mU/L。本方法适用于血清中碱性磷酸酶的测定。测定结果与临床上常用的以对硝基苯磷酸酯为底物的分光光度法相比,无显著性差异。

A new substrate, salicyl phosphate （SP） , was developed for the fluorimetric determination of alkaline phosphatase （ALP）activity. The product of the enzyme reaction is salicylic acid （SA） , which is a strong fluorescent product. The amount of SA generated is proportional to ALP activity. Optimal conditions for the determination of ALP activity were investigated. The linear range and detection limit for the determination were 0.03 -6.00 U/L and 7.04 mU/L, respectively. This method is simple, and can be successfully applied to assess ALP activity in human serum with good accuracy and precision. The results were evaluated by comparing with a standard colorimetric assay using p-nitrophenyl phosphate as ALP substrate