Optimized Densitometric Analysis for Determination of Triterpenoid Isomers in Vitex negundo L. Leaf

Triterpenoid isomers oleanolic acid (OA) and betulinic acid (BA) were densitometrically separated and quantified using an optimized high performance thin layer chromatography analysis in Vitex negundo. Multiple wavelengths (400–675 nm at 25 nm increment) were scanned during the densitometric analysis, and 600 nm was found optimum to quantify OA and BA simultaneously. This technique allowed optimum separation of OA and BA with distinguishing band color and corresponding Rf than the previous reports. The leaf extract contained higher amount of BA (56.74 ± 2.83 ng/band) compared to OA (16.25 ± 0.81 ng/band). The method provides better separation, resolution at optimized wavelength for these compounds at lower concentration levels.