The soluble lignin and furan compounds in hydrolysate of lignocellulose biomass have absorbed peaks and inter-superimpose in UV spectroscopy. Therefore, this paper developed a method for the determination of furfural and 5-hydroxymethyl furfural in ethanol-water refining liquors of lignocellulose biomass based on UV spectroscopy. The study showed that there is a good and linear relationship between maximum absorption (λmax) and the concentration percent of 5-HMF (W5-HMF) of M, the regression coefficient up to 0.998 4 (R2=0.998 4). Furthermore, a better linear relationship was also observed between the concentration of mixture (cM) and absorptivity (A) when the W5-HMF was constant, the R2 of 0.998 6～1.000 0 and reproducibility are desirable. The contents of F and 5-HMF could be calculated accurately according to the perfect liner relationship of A and cM, λmax and W5-HMF, and the relative deviation of F and 5-HMF of 5.6% and 3.7%, recovery of 89%～96%, RSD of 0.21%～0.85%, respectively. Overall, The simple rapid and precise method in this paper can be applied to the determination of the F and 5-HMF in the hydrolysate of ethanol-water refining lignocellulose biomass.