A ratiometric fluorescence probe based on graphene quantum dots and o-phenylenediamine for highly sensitive detection of acetylcholinesterase activity.

Abstract

By using graphene quantum dots (GQDs) and o-phenylenediamine (OPD), a ratiometric fluorescence probe was designed for the highly sensitive and selective detection of AChE. GQDs with strong fluorescence were synthesized by the one-step hydrothermal method. The optimal emission wavelength of GQDs was 450nm at the excitation wavelength of 375nm. MnO2 nanosheets with a wide absorption band of 300-600nm were prepared at room temperature. Because of the extensive overlap between the absorption spectrum of MnO2 nanosheets and the excitation and emission spectra of GQDs, the fluorescence of GQDs at 450nm was efficiently quenched by the inner-filter effect. Meanwhile, due to the peroxidase-like activity of MnO2 nanosheets, OPD was catalytically oxidized to 2,3-diaminophenazine (oxOPD), a yellow fluorescent substance with a new emission peak at 572nm. When AChE was present, the substrate acetylthiocholine (ATCh) was hydrolyzed to thiocholine (TCh) that is capable of decomposing MnO2 nanosheets. Therefore, the quench of GQDs and the oxidation of OPD by MnO2 nanosheets were suppressed, resulting in the fluorescence recovery of GQDs at 450nm, while the fluorescence decrease of oxOPD at 572nm. Utilizing the fluorescence intensity ratio F450/F572 as the signal readout, the ratiometric fluorescence method was established to detect AChE activity. The ratioF450/F572 against the AChE concentration demonstrated two linear relationships in the range 0.1-2.0 and 2.0-4.5mUmL-1 with adetection limit of 0.09mUmL-1. The method was applied tothe detection of positive human serum samples and the analysis of the inhibitor neostigmine. Due to the advantages of high sensitivity, favorable selectivity, and strong anti-interference, the method possesses an application prospect in clinical diagnosis of AChE and the screening of inhibitors. Graphical abstract Schematic presentation of a ratiometric fluorescence method for the detection of acetylcholinesterase (AChE). The fluorescence of graphene quantum dots (GQDs) is quenched and o-phenylenediamine (OPD) is oxidized to generate fluorescent product 2,3-diaminophenazine (oxOPD) by MnO2 nanosheets. When AChE is present, acetylthiocholine iodide (ATCh) is hydrolyzed to thiocholine (TCh) with reducibility for decomposing MnO2 nanosheets. Due to the decomposition of MnO2 nanosheets, the quenching of GQDs and oxidation of OPD are suppressed. The fluorescence of GQDs at 450nm is enhanced, while the fluorescence of oxOPD at 572nm is reduced. The fluorescence intensity ratio F450/F572 is used to establish the ratiometric fluorescence method for AChE activity.