High-performance thin-layer chromatographic–densitometric determination of the major triterpene saponins and their aglycones from Centella asiatica

Abstract

Centella asiatica (L.) Urban, Apiaceae, is a medicinal plant, rooted in the Ayurvedic medicine with wide and potent health beneficial activity, and is of great interest to researchers concerning phytotherapy due to its numerous pharmacological activities. In this paper, we have developed a valid thin-layer chromatography (TLC)—densitometric method for the simultaneous determination of the four main C. asiatica active compounds. The additional goal was to achieve the conditions of determination, which would not require spraying with the derivative reagent and subsequent heating — the factors that greatly deteriorate the validation parameters. In order to do so, we have tested diff erent sorbents (alumina, silica gel, and high-performance thin-layer chromatography [HPTLC] silica gel) and a number of eluents. The best results were obtained with HPTLC silica gel plates eluted with a mixture of CHCl3-MeOH-H2O (10:8:2, v/v) and analysed with a densitometer at λ = 202 nm. The chromatographic method was validated for linearity, limit of detection, limit of quantification, repeatability, intermediate precision, and recovery.