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Determination of the activity of alkaline phosphatase by using nanoclusters composed of flower-like cobalt oxyhydroxide and copper nanoclusters as fluorescent probes

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Abstract

The authors describe a sensitive fluorometric method for the determination of the activity of alkaline phosphatase (ALP). It is based on the use of a composite prepared consisting of flower-like cobalt oxyhydroxide (CoOOH) and copper nanoclusters (CuNCs). On formation of the CuNC-CoOOH aggregates, the fluorescence of the CuNCs is quenched by the CoOOH sheets. If, however, the CoOOH sheets are reduced to Co(II) ions in the presence of ascorbic acid (AA), fluorescence recovers. AA is formed in-situ by hydrolysis of the substrate ascorbic acid 2-phosphate (AA2P) as catalyzed by ALP. Thus, the ALP activity can be detected indirectly by kinetic monitoring of the increase in fluorescence, best at excitation/emission wavelengths of 335/410 nm. The assay allows ALP to be determined in 0.5 to 150 mU·mL−1 activity range and with a 0.1 mU·mL−1 detection limit. The method was successfully applied to the determination of ALP activity in (spiked) human serum samples. The assay has attractive features in being of the off-on type and immune against false positive results.

A fluorescent bioassay is reported for the determination of the activity of alkaline phosphatase (ALP). It is exploiting the ascorbic acid (AA)-induced decomposition of nanoclusters composed of flower-like cobalt oxyhydroxide and copper nanoclusters. ALP catalyzes hydrolysis of ascorbic acid 2-phosphate (AA2P) and dephosphorylation to form AA.

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Acknowledgements

This work was financially supported by National Natural Science Foundation of China (No. U1704153, 21305119), Plan for Young Excellent Teachers in Universities of Henan Province (No. 2017GGJS), Nanhu Scholars Program for Young Scholars of XYNU, Plan for Scientific Innovation Talent of Henan Province (No. 2017JR0016), the Science & Technology Innovation Talents in Universities of Henan Province (No. 16HASTIT004).

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Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, Henan Province Key Laboratory of Utilization of Non-metallic Mineral in the Sourth of Henan, Xinyang Normal University, Xinyang, 464000, People’s Republic of China

    Hai-Bo Wang, Yang Li, Ying Chen & Yan-Ming Liu

  2. College of Life Science, Yantai University, Yantai, 264005, People’s Republic of China

    Zi-Ping Zhang

  3. Institute for Conservation and Utilization of Agro-bioresources in Dabie Mountains, Xinyang Normal University, Xinyang, 464000, People’s Republic of China

    Tian Gan & Yan-Ming Liu

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  1. Hai-Bo Wang
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  2. Yang Li
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  3. Ying Chen
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  4. Zi-Ping Zhang
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  5. Tian Gan
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  6. Yan-Ming Liu
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Correspondence to Hai-Bo Wang or Tian Gan.

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Wang, HB., Li, Y., Chen, Y. et al. Determination of the activity of alkaline phosphatase by using nanoclusters composed of flower-like cobalt oxyhydroxide and copper nanoclusters as fluorescent probes. Microchim Acta 185, 102 (2018). https://doi.org/10.1007/s00604-017-2622-4

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  • DOI: https://doi.org/10.1007/s00604-017-2622-4

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