To establish a method to determine the contents of asenosine in different species of Rehmannia glutinosa.The contents were determined by HPLC, with a column of KYWG-C18 as stationary phase, 6% ace tonitrile as mobile phase and detecting wavelength at 260 nm.Adenosine has a good linearity in the range of 0.002 mg/ml-0.01 mg/ml, Y = 1.7742 x 10(-4) + 8.9021 x 10(-7) X, r = 0.9995. The average recovery is 94.1% and RSD = 1.86% (n = 5).The method is fast, reliable and simple.