Quantitative estimation of betulin in different plant parts of Dilleniapentagyna Roxb. through validated HPTLC method

Abstract

A new, simple and accurate high-performance liquid chromatographic (HPTLC) method was developed for estimation of betulin (BE) in fruits, leaves, root bark and stem bark of Dillenia pentagyna , a RET species of Madhya Pradesh state of India. HPTLC aluminum plates precoated with 0.2 mm layer of silica gel 60F 254 were used to separate BE from the plant samples using the mobile phase, petroleum ether: ethyl acetate: acetonitrile (8: 2: 0.1, v/v/v). The anisaldehyde - sulphuric acid reagent was used to derivatize the plate followed by heating at 105°C on CAMAG HPTLC hot plate for 5 minutes. The peak corresponding to reference compound BE and that of the extract appeared at the same R F 0.21. Image of the plate was captured and HPTLC scan was performed densitometrically at ƛ max = 580 nm under max tungsten lamp in CAMAG TLC Scanner4 equipped with win CATS software. The developed HPTLC method was validated as per norms of the International Conference on Harmonization (ICH) for linearity, sensitivity, precision, accuracy and specificity. The BE content in fruit, leaf, root bark and stem bark samples of D. pentagyna was found as 0.101 ± 0.03%, 0.091 ± 0.09, 0.368 ± 0.07 and 0.440 ± 0.11% respectively.