A sensitive, selective and robust qualitative and quantitative densitometric high-performance thin layer chromatographic method was developed and validated for the determination of iridoid glycoside in the aerial part of Gambhari (
Gmelina arborea). Iridoid gycoside 6-
O-(2″,3″-dibenzoyl)-
-
l-rhamnopyranosylcatalpol (IG) was used as a chemical marker for the standardization of
G. arborea plant extracts. The separation was performed on aluminum Kieselgel 60F
254 TLC plates using chloroform–methanol as mobile phase. The quantitation of IG was carried out using the densitometric reflection/absorption mode at 240 and 430 nm after post-chromatographic derivatization with vanillin–sulphuric acid reagent. A precise and accurate quantification can be performed in the linear working concentration range of 1000–5000 ng/spot with good correlation (
r2 = 0.994). The method was validated for peak purities, precision, robustness, limit of detection (LOD) and quantitation (LOQ), etc., as per ICH guidelines. Specificity of quantitation was confirmed using retention factor (
Rf), UV–vis spectral correlation and ESI-MS spectra of marker compound (IG) in sample track.