Abstract: Phytosterols can effectively reduce the cholesterol content in human serum.But the solubility of free phytosterols is poor. To improve the liposolubility of phytosterols, esterification with fatty acid is normally adopted. Soybean sterol conjugated linoleate was analyzed quantitatively and qualitatively with normal phase high performance liquid chromatography. Influences of variables, such as the molar ratio of alcohol to acid, catalyst loading, reaction temperature and reaction time, on the yield of soybean sterol conjugated linoleate were investigated by single factor test and orthogonal design. The optimum conditions were as follows: CRL catalyst amount 6%, molar ratio of alcohol to acid 1∶3, reaction temperature 50 ℃ and reaction time 48 h. Under these conditions, the yield of soybean sterol conjugated linoleate was 98.94%. The synthesized product was then purified and characterized by infrared spectroscopy. In this paper, highly purified soybean sterol conjugated linoleate product was synthesized by lipase-catalyzed method, providing a theoretical guidance and technical support for the development and high value utilization of new resource food.