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.
Authors:Danuta Raj, Adam Kokotkiewicz and Maria Łuczkiewicz
An optimized chromatographic method has been established for separation of alkaloid fractions in the herb
and in different types of in-vitro cultures. TLC separation of
indolizidine alkaloids was optimized by using four types of plate, both silica gel (Si60 F
TLC plates, and Si60 F
and LiChrospher Si60 F
HPTLC plates) and reversed-phase (RP 18W), with a range of solvent mixtures as mobile phases. The best resolution was achieved with HPTLC Merck LiChrospher Si60 F
and chloroform-methanol 100:5 (
) as mobile phase. The method, with densitometric detection, was then used for quantitative analysis of the securinine and allosecurinine content of extracts from different
samples. The method was validated for linearity, limits of detection and quantification, repeatability, intermediate precision, and recovery.