Abstract: A HPLC separation and determination of sarmentosine in the plant and its preparations is reported. The method is proceeded with reverse phase chromatographic system using ODS column in series with amino bonded precolumn to separate sarmentosine from the major interfering parts. The mobile phase used is methanol acetonitrile-water(2:6:100) and the active component is detected with UV detector at λ 225 nm. A procedure of directly introducing the tissue extract into the chromatographic system has been proposed which is intended to be used as a quick analytical method to meet the requirements in investigating plant resource distribution. The method is suitable for the determination of sarmentosine in fresh plant extracts, crude drugs and various medical preparations. The relationship between the sarmentosine content and the variations in growing district, species and its collecting time has been reported. The method can also be used in quality control during processing. It is also possible to study the reason of inconstancy of clinical effects in medical preparations.