Fluorometric turn-on determination of the activity of alkaline phosphatase by using WS2 quantum dots and enzymatic cleavage of ascorbic acid 2-phosphate.

Abstract

A method is described for the determination of the activity of alkaline phosphatase (ALP). It is based on the reversible modulation of the fluorescence of WS2 quantum dots (QDs). The fluorescence of the QDs is quenched by Cr(VI) but restored by free ascorbic acid (AA). The detection scheme relies on the fact that ALP hydrolyzes the substrate ascorbic acid 2-phosphate to produce AA, and that enzymatically generated AA can restore the fluorescence of the QDs. The signal (best measured at excitation/emission peak wavelengths of 365/440 nm) increases linearly in the 0.5 to 10U·L-1 ALP activity range, with a detection limit of 0.2U·L-1. The method was applied to the determination of ALP activity in human serum samples and demonstrated satisfactory results. Graphical abstract The fluorescence of chromate-loaded tungsten disulfide quantum dots (QDs) is quenched but restored after reaction with ascorbic acid that is formed by the catalytic action of alkaline phosphatase (ALP) on ascorbic acid 2-phosphate (AAP). The increase in fluorescence can be related to the activity of ALP.