HPLC determination of oleanolic acid and ursolic acid in Hedyotis diffusa Willd from different sources

Objective To determine the oleanolic acid and ursolic acid in Hedyotis diffusa Willd from different sources and to provide a reference for quality control of raw materials and preparations. Methods The analysis was performed by HPLC with C18 column. The mobile phase consisted of methanol-water-glacial acetic acid-triethylamine (91.5∶8.5∶0.04∶0.1) with detection wavelength at 210 nm, the flow rate was 0.80 ml/min and the column temperature was maintained at 20 ℃. Results The linear range of oleanolic acid and ursolic acid were 20.4～326.4 μg /ml (r= 0.9991) and 39.2～627.2μg /ml (r=0.9994) respectively. The average recovery of oleanolic acid and ursolic acid were 98.0～99.0%, with RSD being 0.87% and 1.22% respectively. Conclusion The method developed in this study appeared accurate, convenient and reproducible and can be used for quality control of oleanolic acid and ursolic acid raw materials or preparations of Hedyotis diffusa Willd.