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

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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.

Schematic presentation of the nitrogen and boron co-doped carbon dot-based fluorometric method for determination of alkaline phosphatase (ALP) activity.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (21705056), the program for Young Taishan Scholars of Shandong Province, the Natural Science Foundation of Shandong Province (ZR2017MB022, ZR2018BB057 and ZR2018PB009) and the start-up funding from University of Jinan (511-1009408, 511-1009424).

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Authors and Affiliations

  1. School of Materials Science and Engineering, University of Jinan, Jinan, 250022, People’s Republic of China

    Pengjuan Ni, Chuanxia Chen, Yuanyuan Jiang, Yizhong Lu & Xun Hu

  2. College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes (Ministry of Education), Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Institute of Molecular and Nano Science, Shandong Normal Univeristy, Jinan, Shandong, 250014, People’s Republic of China

    Junfeng Xie

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Ni, P., Xie, J., Chen, C. et al. Fluorometric determination of the activity of alkaline phosphatase and its inhibitors based on ascorbic acid-induced aggregation of carbon dots. Microchim Acta 186, 202 (2019). https://doi.org/10.1007/s00604-019-3303-2

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