Molecular dynamics (MD) simulation provides a quantitative method for screening efficient optical clearing agents. Some excellent optical clearing agents were obtained through the MD simulation of alcohols with hydroxyl groups. Among of the agents, fructose, one of polyols has the better efficacy which is one of monoaccharides. In comparison, disaccharides received little attention. In this study, sucrose and maltose were screened from common disaccharides for comparing with fructose. Actually, each agent has different saturation concentration. Here, the combination of molecular dynamic simulation and in vitro experiment was employed in this study. Firstly, the hydrogen bonds were used to predict the optical clearing potential of sucrose, maltose and fructose by using molecular dynamic simulation, respectively. The simulated results indicated that the optical clearing potential in a descending order as: sucrose, maltose and fructose. The reduced scattering coefficient of in vitro rat skin samples was used to evaluate the skin optical clearing potential based on measurements of reflectance and transmittance using a commercial spectrophotometer with an integrating sphere. Both the experimental and simulated results show that for the same concentration, disaccharides have better optical clearing potential than fructose. However, maltose has too low saturation concentration to produce similar optical clearing efficacy as the other two. By contrast, sucrose is the best one even though it has lower saturation concentrations than fructose.