Fluorometric determination of the activity of alkaline phosphatase and its inhibitors based on ascorbic acid-induced aggregation of carbon dots

Mikrochimica Acta
Pengjuan NiXun Hu

Abstract

The authors describe a fluorometric method for determination of the activity of alkaline phosphatase (ALP) and its inhibitors. Nitrogen and boron co-doped carbon dots (C-dots) with excitation/emission peaks at 490/540 nm act as the fluorescent probe. The C-dots were prepared by hydrothermal carbonization starting from 3-aminophenylboronic acid as the sole precursor. On the basis of the boronic acid-triggered specific reaction with cis-diols, the boronic acid modified C-dots can bind to ascorbic acid that is generated by ALP-catalyzed hydrolysis of ascorbic acid 2-phosphate. This results in particle aggregation and quenching of fluorescence. If the ALP inhibitor Na3VO4 is introduced into the system, the activity of ALP is reduced and the fluorescence of C-dots recovers. This fluorometric method allows for the determination of ALP activity in the range from 0.2 to 6.0 mU mL-1 with a detection limit of 0.16 mU mL-1. The IC50 value for the inhibitor Na3VO4 is 3.6 μM. The method is convenient and cost-effective. It does not require complicated operations and in our perception widens the scope of applications of C-dots in bioanalytical sciences. Graphical abstract Schematic presentation of the nitrogen and boron co-doped carbon dot-b...Continue Reading

References

Aug 6, 2013·Accounts of Chemical Research·Changqin DingYang Tian
Oct 23, 2015·Organic & Biomolecular Chemistry·Mariia MiliutinaPeter Langer
Jan 15, 2016·ACS Applied Materials & Interfaces·Adeline Huiling LooMartin Pumera
Sep 15, 2016·Advanced Materials·Huijun YuTierui Zhang
Apr 4, 2017·Luminescence : the Journal of Biological and Chemical Luminescence·Meijuan LiangXingguo Chen

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