Photoluminescent C-dots@RGO probe for sensitive and selective detection of acetylcholine

We have developed a sensitive and selective photoluminescence (PL) quenching assay for the detection of acetylcholine (ACh) that uses reduced graphene oxide decorated with carbon dots (C-dots@RGO). The highly stable C-dots@RGO synthesized from catechin and graphene oxide through a hydrothermal reaction displays excitation-wavelength dependence of PL. Acetylcholinesterase (AChE) converts ACh to choline, which in turn is oxidized by choline oxidase (ChOx) to produce betaine and H2O2 that generates the reactive oxygen species (ROS). The as-produced ROS induces PL quenching of the C-dots@RGO through an etching process. With respect to sensitivity, the optimal reaction/sensing temperature and pH are 37 °C and 9.0, respectively, using C-dots@RGO (0.4 mg·mL(-1)) and AChE and ChOx at the activities of 0.5 and 0.1 unit·mL(-1), respectively. The PL intensity (excitation/emission wavelengths 365/440 nm) of the C-dots@RGO is inversely proportional to the concentration of ACh over a range of 0.05-10 nM (r = 0.997), with a limit of detection (signal-to-noise ratio 3) of 30 pM. We have validated this assay by determination of concentrations of ACh in plasma and blood samples, with results of 2.6 ± 0.8 nM (n = 5) and 6.8 ± 0.4 nM (n = 5), respectively. Our study opens an avenue for the detection of various analytes by use of C-dots@RGO in conjunction with different enzymes, substrates, and/or inhibitors.