- 作者：Xiaojing Zhu ; Chongqiu Jiang
- 关键词：Alkaline phosphatase ; Fluorimetry ; 8-Quinolyl phosphate
- 刊名：Clinica Chimica Acta
- 出版年：2007
- 期刊代码：54_00098981
- 类别：med
- 出版时间：2 February 2007
- 卷：377
- 期：1-2
- 页码：150-153
- 文件大小：250 K
Alkaline phosphatase (ALP) is an important target for clinical analysis. 8-Quinolyl phosphate (QP) was developed as a new substrate for the fluorimetric determination of ALP activity.
Methods
QP is a strong fluorescent substance and the product of the enzyme reaction is 8-hydroxyquinoline (HQ), which has no fluorescence. Under the optimal conditions for the determination of ALP, the decreased fluorescence intensity via the enzyme reaction is proportional to ALP activity. The fluorescence intensity was measured at λex/λem = 318/495 nm before and after the enzyme reaction.
Results
QP reacted with ALP in the buffer solution of pH = 9.5 and incubated for 20 min at 37.0 °C were selected as the optimal conditions for the determination of ALP. The linear range and detection limit for the determination of ALP are 1.0–16.0 and 0.229 U/l, respectively. With this method, ALP could be applied to assess ALP in human serum and the results were evaluated by comparison with a standard colorimetric assay using p-nitrophenyl phosphate as ALP substrate.
Conclusions
This method is simple, practical and can be used as an alternative to assess ALP in clinical analysis.