Dual-signal uric acid sensing based on carbon quantum dots and o-phenylenediamine

Fluorescent carbon quantum dots (CQDs), which showed excitation-dependent emission characteristics, were prepared using a facile hydrothermal method. The structure and optical properties of CQDs were characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, UV-Vis spectroscopy, and fluorescence spectroscopy. These CQDs also showed peroxidase-like activity and could catalyze the H₂O₂-mediated oxidation of o-phenylenediamine (OPD) to form 2,3-diaminophenazine (DAP) with an absorption peak at 420 nm. DAP exhibited an obvious fluorescence emission at 550 nm under the excitation of 360 nm. On the other hand, it decreased the fluorescence of CQDs at 450 nm via inner filter effect. The experimental results indicated that the H₂O₂ concentration affected the color of DAP and the fluorescence intensity of CQDs and DAP. Thus, a colorimetric and ratiometric fluorescence dual-signal method was established for measuring the concentrations of H₂O₂ and uric acid (UA). The effects of pH, incubation temperature, incubation time, and OPD concentration on the response were investigated. Under the conditions of pH 7.5, temperature 50 °C, incubation time 30 min, and OPD 1.5 mM, the absorbance and fluorescence intensity ratio responses were linearly dependent on UA concentration ranging from 5.0 μM to 100 μM. The limits of detection were 0.7 and 0.5 μM with a colorimetric method and ratiometric fluorescence method, respectively. More importantly, this dual responsive method has been applied to the determination of UA in urine samples with satisfactory results.