Objective To study the role of mestinon-phospholipid complex in promoting the intestinal absorption of mestinon in rats. Methods The mestinon-phospholipid complex was prepared and its composition rate, physico-chemical property and N-octanol/water partition coefficient were examined. Six healthy male rats were randomly divided into two groups. Using the rat model of intestinal absorption, we studied the intestinal absorption rate constant and effective permeability of mestinon-phospholipid complex and free mestinon. Results The composition rate of mestinon-phospholipid complex was (84.02±1.68)%. The characteristic endothermal peak of mestinon disappeared in the DSC curve of mestinon-phospholipid complex. The phase transition temperature of the complex was lower than that of phospholipids, the ultraviolet spectrum profile of the complex was similar to that of mestinon, and the infrared spectrum of the complex was significantly different from that of the physical mixture of mestinon and phospholipid. The particle size of mestinon-phospholipid complex was about (204.6±0.3) nm,and its zeta-potential was (-25.12±0.16) mV. The particle size was largely in a normal distribution. Both the coefficient of recovery and precision of mestinon in intestinal circulating fluid met the requirement. The N-octanol/water partition coefficient of mestinon-phospholipid complex was about 17 folds that of mestinon. The absorption rate constant and the effective permeability of mestinon-phospholipid complex were 2.32 and 2.52 folds those of free mestinon, respectively. Conclusion The mestinon-phospholipid complex can greatly improve the intestinal absorption of mestinon in rats.