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Although carbon dots (CDs) have been widely applied in nanobiotechnology and biomedicine, few studies have evaluated the intrinsic self-bioactivities and no reports have described the disaccharidase inhibitory activities of CDs. In this study, charred Fructus crataegi -derived CDs (CFC-CDs) having an average diameter of 1.3–5.6 nm are developed, without further modifications. Owing to an abundance of surface groups, CFC-CDs show distinct solubility, biocompatibility, and bioactivity. Moreover, CFC-CDs inhibit the catalytic activities of sucrase and maltase in vitro and reduce postprandial blood glucose levels in vivo , possibly by acting as a disaccharidase inhibitor. This discovery provides guidance for further research on the bioactivities of CDs and supports their potential applications in the biomedical and healthcare fields.