Abstract : The objective of the current study is to develop a quantitative method for the simultaneous evaluation of major active components of Pongamia pinnata seed oil – Pongamol and Karanjin - by High-Performance Thin-Layer Chromatography (HPTLC). Chromatographic separations of the active compounds were achieved by silica gel 60 F254 plates utilizing a mobile phase of petroleum benzine:ethyl acetate:formic acid (8:2:0.1, v/v/v), along with densitometric detection at a wavelength of 366 nm. The newly developed method of analysis displayed excellent linearity, and correlation coefficients R² = 0.9893 for pongamol and 0.9926 for karanjin, among the concentration ranges of 0.2–1.0 μg/spot and 0.1–2.0 μg/spot, respectively. Less than 2% RSD values for precision with intra- and inter-day proved the reliability of the developed method. Accuracy of the method was confirmed by the recovery rates of 98.5–101.4% for pongamol and 97.9–102.2% for karanjin. The LOD obtained was 0.0238 μg/spot (pongamol) and 0.0279 μg/spot (karanjin), and the LOQ was 0.0721 μg/spot and 0.0848 μg/spot, respectively. This method provides an easy tool for quantification of active compounds in herbal formulations and in the development of cosmeceutical products.

Keyword : HPTLC, Karanjin, Method development, Pongamia pinnata, Pongamol.