The invention relates to a preparation method for single-layer molybdenum disulfide nanosheets. The method comprises the following steps: (1) 200-250mg of molybdenum disulfide powder with a particle size less than 2 micrometers, 90-110mg of chitosan powder, 1-1.2g of an ionic liquid and 10-15mL of an organic solvent are uniformly mixed and transferred to a sealed glass container, so a first dispersion solution is obtained; (2) the first dispersion solution is subjected to ultrasonic treatment, after the ultrasonic treatment is finished, high-speed centrifugation separation is performed, and a precipitate is collected; (3) the precipitate obtained in the step (2) is dispersed in an acetic acid solution with a concentration of 0.5%, centrifugation separation is performed, a precipitate is collected, and the above process is repeated for two times; and (4) the precipitate obtained in the step (3) is dispersed in deionized water, low-speed centrifugation separation is performed, and a supernatant is collected, so a single-layer molybdenum disulfide nanosheet solution is obtained. According to the invention, the process is simple, costs are low, the yield is higher than 25%, repeatability is good, and defects of large size, a low single-layer yield, nonuniform particle size and the like of molybdenum disulfide nanosheets obtained in the prior art can be overcome.