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Carbon dots without heteroatoms-doping and surface modifications were designed to be a novel chemosensing strategy on the quantitative detection of uric acid (UA) with the aid of uricase-induced enzymatic reaction and Fenton reaction. In this work, ascorbic acid (AA)-derived carbon dots (A-CDs) were prepared in the mixture of ethanol and water via one-step hydrothermal synthesis at a relatively low temperature (120 °C) for 10 h. The resultant A-CDs were proved to be excitation-independent. When excited at the wavelength of 420 nm, the nanodots displayed green fluorescence (535 nm) which was then linearly quenched as UA concentration increased in the range of 0–56 μM, according to which the detection limit was calculated to be 0.49 μM. With regards to the excellent sensitivity and selectivity to UA, real sample assay was performed on the A-CDs detection system, which provided relatively reliable recoveries of UA contained in human serum/urine. Besides, in view of the high quantum yield, the A-CDs were applied to live-cell imaging assay and were considered to become an alternative tracer tool in biomedical imaging. [Display omitted] • Carbon dots are prepared at a relatively low temperature without heteroatoms doping and surface modification. • The sensing system based on the carbon dots has a low detection limit to uric acid. • The system not only allows for quantifying uric acid and H 2 O 2 , but also possesses the imaging ability to living cells. [ABSTRACT FROM AUTHOR]