The invention relates to a method for determining the concentration of stable dispersion water solution C60. The method comprises the steps of firstly, preparing different concentrations of C60-Toluene standard solution, determining the absorbance at the wavelength of 334-336nm, performing linear regression to the concentration values and corresponding absorbance value, thus obtaining a standard curve; then taking stable dispersion water solution C60 to be determined, adding calcium chloride solution and toluene, wherein the ratio of the stable dispersion water solution to calcium chloride solution to toluene is 1:(0.5-1):(0.5-2); sealing, performing ultrasonic dispersion for 10-15min to form white emulsion, oscillating and standing to obtain a two-layer solution, freezing for 30-60min in the condition of minus 20 DEG C so as to entirely condense the water phase to be in a solid state and keep an organic phase under a liquid state; entirely absorbing the organic phase solution, determining the absorbance, and calculating the concentration of the C60 according to the standard curve; repeating the steps, adding the concentrations of the C60 obtained by twice extraction, thus obtaining the final determined concentration of the stable dispersion water solution C60 to be determined. The method is simple and fast to operate, and capable of performing batch determination.