The invention provides an N, P and S-codoped fluorescent carbon quantum dot and a preparation method and application thereof, and belongs to the technical field of luminescent nanomaterials. The preparation method comprises the steps that a biological bacterium (saccharomycete or escherichia coli or staphylococcus aureus or aspergillus niger) is taken as a carbon source, water of the corresponding volume is added, and a hydrothermal reaction is performed to obtain a dark brown solution; after the reaction stops and a reaction kettle is naturally cooled, dialysis is performed to remove impurities, an aqueous carbon quantum dot solution is obtained, and after freeze drying is performed, the N, P and S-codoped fluorescent carbon quantum dot is obtained. According to the method, the biomass is taken as the carbon source, the raw material is wide in source, low in cost, green and environmentally friendly, the technology is simple, and the preparation condition requirement is low. The obtained carbon quantum dot can be applied to detection on Cr (VI) ions, MnO4<-> and ascorbic acid by serving as a switch-type fluorescent probe and also can be used for cell imaging.