The invention relates to a method for preparing monolayer molybdenum disulfide quantum dots based on electronic dynamic regulation and control, in particular to a method for obtaining the monolayer molybdenum disulfide quantum dots with uniform particle sizes by obtaining molybdenum disulfide suspension and then centrifugally separating and belongs to the field of femtosecond laser application. The method disclosed by the invention comprises the following steps: aiming at the characteristics of a molybdenum disulfide material, carrying out pulse shaping on traditional femtosecond laser monopulse by using a Michelson interferometer to form a pulse sequence; adjusting energy of the pulse sequence, delay among sub pulses, laser scanning speed and scanning intervals; ablating blocky molybdenum disulfide in water and further regulating and controlling local transient electronic dynamics in the interaction process of laser and the material to form multistage photoablation monolayer molybdenum, and obtaining the monolayer molybdenum disulfide quantum dots with the uniform particle sizes; carrying out laser-induced water dissociation to enhance light absorption and improve the yield of the molybdenum disulfide quantum dots. The method disclosed by the invention has the advantages of no need of special chemical environment or chemical reagent, greenness, no pollution, and simple and flexible operation.