Abstract: An accurate HPLC method for the determination of betulin content from the bark of Betula platyphylla Suk.was described. The analytical column ODS C18 was used,with mobile phase methanol:water(88:12) using UV (210 nm),column temperature 30℃,column pressure 6.4 MPa,flow rate 1 mL/min.The linear range of betulin was (15.0-300.0 μg).Average recovery rate was 93.93%.Relative standard deviation of one sample repeated for 5 times was 3.26%.Compared with colorimetric method and GC method,HPLC method is more accurate and convenient.