Ursolic acid and Oleanolic acid are an anti-cancer and anti-inflammatory pentacyclic triterpenoids, an active constituent of plant Plumeria obtusa. Ursolic acid and Oleanolic acid content in Plumeria obtusa has not been reported till date. Thus HPTLC method was developed for quantification of Ursolic acid and Oleanolic acid in leaves of Plumeria obtusa. The separation of Ursolic acid and Oleanolic acid were performed on aluminum backed silica gel G 60 F254 as stationary phase. Mobile phase system optimized was Hexane: Ethyl acetate: Methanol (8.2: 1.8: 0.5 v/v/v) with pre-saturation time of 45 min. Spectrophotometric evaluation was performed at 540 nm. Ursolic acid and Oleanolic acid are chemical isomers and they not separated as such. Ursolic acid and Oleanolic acid were resolved at Rf 0.28 ± 0.03 and 0.41 ± 0.03 respectively by pre chromatographic derivatization with iodine. The polynomial regression of method was found to be within range 400-1400 ng/spot with mean percentage recovery of 97.59 ± 0.99% for ursolic acid and 1200-4200 ng/spot with mean percentage recovery of 96.61 ± 0.83% for Oleanolic acid. The amount of Ursolic acid and Oleanolic in leaves were found to be 0.197 % and 0.0253 % respectively.
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