Authors:Prawez Alam, Y.T. Kamal, Mohammed H. Alqarni, Hala H. Zaatout and Maged S. Abdel-Kader
A new rapid, simple, economical, and environment-friendly reversed- phase high-performance thin-layer chromatography (RPHPTLC) method has been established for the simultaneous determination of glycyrrhizin and glabridin in Glycyrrhiza glabra roots, rhizomes and selected herbal formulations. The method was carried out using RP-18 silica gel 60 F254S HPTLC glass plates and methanol–water (7:3 v/v) as the mobile phase. The developed plates were scanned and quantified densitometrically at 256 and 233 nm for glycyrrhizin and glabridin, respectively. Glycyrrhizin and glabridin peaks from G. glabra roots and rhizomes and herbal formulations were identified by comparing their single spots at RF = 0.63 ± 0.02 and RF = 0.28 ± 0.01, respectively. Linear regression analysis revealed a good linear relationship between the peak areas and the amounts of glycyrrhizin and glabridin in the ranges of 1000–7000 and 100–700 ng band−1, respectively. The method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines for precision, accuracy, and robustness. The proposed method will be useful to determine the therapeutic doses of glycyrrhizin and glabridin in herbal formulations as well as in bulk drug.
The merits of chemometrics in categorizing different Egyptian olive chemovarieties based on their compositional integrity were implemented in this study. Fingerprints of 9 different olive leaves varieties cultivated in Egypt were established using reversed-phase high-performance thin-layer chromatography (RP-HPTLC) prior to and after post-chromatographic derivatization with natural product-polyethylene glycol (NP/PEG) reagent and image analysis using ImageJ® software in order to build 2 separate data matrices. The chromatographic fingerprints were separately subjected to unsupervised pattern recognition multivariate analysis to build 2 separate models using principal component analysis (PCA) and hierarchical clustering analysis (HCA) algorithms to explore the distribution pattern of different chemovarieties. The second model which involved olive samples’ fingerprints after post-chromatographic derivatization exhibited greater ability to reveal a broader spectrum of phytoconstituents with enhanced sensitivity. Densitometric RP-HPTLC quantification of oleuropein marker was compared to image analysis approach using Sorbfil TLC Videodensitometer® by newly developed and validated methods. Densitometry exhibited better performance characteristics than image analysis method and therefore was executed for determination of oleuropein concentration in the 9 Egyptian olive varieties. Oleuropein marker solely was found to be inadequate for standardization of olive leaves varieties. This study demonstrated a comprehensive approach for the rapid classification of different Egyptian olive varieties, which is crucial to warranting their chemical-consistency and, thereafter, effective consistency.