This paper developed a method for the determination of S-Adenosyl-L-methionine in fermentation liquid using the high performance liquid chromatography coupled with UV detector (HPLC-UV). A good linearity was obtained in the range of 0.1~1.0g/L (r = 0.9969) for S-Adenosyl-L-methionine. The average recoveries of S-Adenosyl-L-methionine were 99.89~101.7% and the relative standard deviations were 0.48~1.36%.The method is simple and rapid with reproducibility for the determination of S-Adenosyl-L-methionine in fermention liquid.