A ratiometric fluorescent biosensor for sensitive determination of α-glucosidase activity and acarbose based on N-doped carbon dots
In this work, a novel ratiometric fluorescent platform for α-glucosidase (α-glu) and its inhibitor was constructed based on N-doped carbon dots (N-CDs). The presence of α-glucosidase can catalyze the release of hydroquinone (HQ) from α-arbutin. Then, the generated HQ can be oxidized and copolymerized with polyethyleneimine (PEI) to form yellowish green fluorescence copolymer (PHQ-PEI) with intense fluorescence emission at 510 nm. When the PHQ-PEI was formed, blue fluorescence of N-CDs at 425 nm was decreased, whereas the fluorescence of PHQ-PEI at 510 nm increased sharply as a result of the fluorescence resonance energy transfer (FRET) effect between N-CDs and PHQ-PEI. However, in the presence of acarbose, the activity of α-glucosidase is inhibited, and α-arbutin can’t be hydrolyzed to hydroquinone, leading to the fluorescence recovery of N-CDs at 425 nm and the fluorescence decreasing of PHQ-PEI at 510 nm. The linear range from 0.2 to 1.6 mU mL-1 and 25-150 μmol L-1 were obtained for α-glucosidase and acarbose detection, respectively. And the detection limit (LOD) for α-glucosidase and acarbose were as low as 0.082 mU mL-1 and 14.5 μmol L-1. Thus, a ratiometric fluorescent sensor with good sensitive and high specificity was established for α-glucosidase assay and satisfactory results were acquired in real sample determination.