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. 2025 Aug 8;192(9):569.
doi: 10.1007/s00604-025-07417-1.

In situ formation of fluorescent non-conjugated polymer dots for α-glucosidase activity assay and inhibitor screening

Feifei Wang  1 , Xinling Wang  1 , Yu Wang  2 , Yin Zhang  1 , Tianxiang Liu  3 , Jian Sun  4
Affiliations

In situ formation of fluorescent non-conjugated polymer dots for α-glucosidase activity assay and inhibitor screening

Feifei Wang et al. Mikrochim Acta. .

Abstract

A novel and facile fluorescence turn-on assay for detecting α-glucosidase (α-Glu) activity and screening inhibitors is reported. This assay exploits α-Glu-catalyzed hydrolysis of L-ascorbic acid-2-O-α-D-glucopyranosyl (AA2G) to release ascorbic acid (AA), triggering an in situ fluorogenic reaction with N-methylethylenediamine (N-MEDA) that generates non-conjugated polymer dots (NCPDs). These NCPDs emit blue fluorescence at 425 nm upon excitation at 350 nm, and the fluorescence intensity increases proportionally with the concentration of α-Glu, enabling quantitative detection of enzymatic activity. The fluorescent assay demonstrates a linear response ranging from 0.01 to 10 U/mL and achieves a low detection limit of 0.0034 U/mL, surpassing most existing methods in sensitivity and width of linear range. In addition, the proposed method has been successfully applied to screen α-Glu inhibitors (AGIs) and determine α-Glu activity in real samples, yielding satisfying results. This work introduces a robust analytical framework and promising approach for clinical diagnosis and anti-diabetic drug discovery.

Keywords: Fluorescence assay; Inhibitor screening; Non-conjugated polymer dots; α-Glu.

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Conflict of interest statement

Declarations. Conflict of interest: The authors declare no competing interests.

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