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A redox modulated ratiometric fluorometric method based on the use of dual-color carbon dots for determination of the activity of enzymes participating in ascorbic acid-related reactions

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Abstract

A turn-on ratiometric fluorescent assay is described for the determination of the activity of enzymes participating in ascorbic acid-forming reactions. Blue-emitting carbon dots (bCDs; with excitation/emission wavelength at 380/450 nm) serve as fluorescent indicator. Their fluorescence is reduced by Fe3+ ions via an inner filter effect. Yellow-emitting CDs (yCDs; with excitation/emission wavelength at 380/550 nm) serve as internal reference because their fluorescence is insensitive to Fe3+. Upon exposure to ascorbic acid (AA), Fe3+ is reduced to Fe2+. Hence, the fluorescence of the bCDs is restored. Thus, enzymes participating in AA-related reactions such as α-glucosidase (α-Glu) and alkaline phosphatase (ALP) can be determined. α-Glu activity was quantified in the range from 0.13 to 6.70 U mL−1, and ALP activity was determined between 2.0 and 130 U L−1. Endowed with excellent sensitivity, selectivity and low background signals, the method may also be used to screen the inhibitors of α-Glu and ALP.

Schematic representation of a redox modulated ratiometric fluorometric method based on the use of dual-color carbon dots for determination of the activity of enzymes participating in ascorbic acid-related reactions. Blue-emitting carbon dots (bCDs) serve as fluorescent indicator while yellow-emitting CDs (yCDs) serve as internal reference.

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (No. 61775099, 21705080) and Natural Science Foundation of Jiangsu Province (No. BK20171487, BK20171043), Science and Technology Development Fund of Nanjing Medical University-Major Project (No.NMUD2018004), and R&D fund for Smart Health Technology Innovation of Nanjing Medical University and Jiangsu Salt Group (No. NMU-SY201801).

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Authors and Affiliations

  1. School of Pharmacy, Nanjing Medical University, Nanjing, Jiangsu, 211166, People’s Republic of China

    Xia Cheng, Guanhong Xu, Fangdi Wei, Yuying Chai, Qin Hu & Yao Cen

  2. Department of Laboratory Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 211166, People’s Republic of China

    Jian Xu & Lin Wang

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  1. Xia Cheng
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Cheng, X., Xu, J., Wang, L. et al. A redox modulated ratiometric fluorometric method based on the use of dual-color carbon dots for determination of the activity of enzymes participating in ascorbic acid-related reactions. Microchim Acta 186, 818 (2019). https://doi.org/10.1007/s00604-019-3820-z

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