High Performance Liquid Chromatography (HPLC) method is a potent analytical technique for determining phytoconstituents even in minute quantities particularly that are generally present in traces concentrations in medicinal plants. The concentration of oleanolic acid (OA) in the Gentiana olivieri Griseb.(GOG) flowers is quantitatively determined by a simple, precise and accurate HPLC method. HPLC assay was performed on reversed-phase C18 column and compound was detected at 210 nm with a flow rate of 1.0 ml/min. The mobile phase consisted of methanol (A) and 0.03 mol/L phosphate buffer, pH 2.9 (B) with a ratio of 85:15 (A: B v/v).The method showed good precision and accuracy with overall intra-day and inter-day variation of 0.36-1.33% and 0.43-1.15%, respectively, and overall recoveries of 99.32%. Application of these methods to determine the OA in Gentiana olivieri Griseb showed that the concentration of OA was found more in the flower as compared to the whole aerial part (average amount in flower 1.82 mg/g DW and 0.75 mg/g DW in whole aerial plant). A simple and accurate HPLC method has been developed for quantitative determination of OA in GOG flowers.