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  • Author or Editor: Karuna Shanker x
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Dikamali (Gardenia lucida Roxb., Rubiaceae family) is found to possess various ethno-medical properties, viz., anti-inflammatory, anthelmintic, cleaning foul ulcers, and wounds, to keep away flies from wounds in animals. In spite of having various medicinal properties, no validated analytical method has been reported so far for the quality assessment of Dikamali. Thus, a high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous quantification of six polymethoxyflavones (PMFs) isolated from G. lucida. Chemical markers, e.g., gardenin-E (1), gardenin-D (2), xanthomicrol (3), 5-desmethynobiletin (4), gardenin-A (5), and gardenin-B (6), of G. lucida gum were selected for quality assessment. For the quantitative determination of the marker compounds, silica gel 60 F254 TLC plates and solvent system n-hexane‒diethyl ether‒1,4-dioxane (4:6:1, v/v) were used for optimum separation and selective evaluation. Densitometric determination was done at 335 nm for the targeted PMFs (1–6). Optimized chromatographic conditions provide well-separated compact bands for the tested polymethoxyflavones. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.06 to 0.10 μg and 0.21 to 0.32 μg, respectively. Curves were linear from 0.33 to 1.67 μg spot−1 with acceptable correlation coefficients ranging from 0.9972 to 0.9988 of all PMFs. The percent mean recoveries of the PMFs ranged from 96.01 to 99.30. The analysis of three G. lucida samples from different locations verified the method to be reproducible and convenient. The method was validated as stated by the International Conference on Harmonization (ICH) guidelines. The developed HPTLC method is found to be accurate and precise. The same has successfully been applied for the estimation of the targeted PMFs in G. lucida gum samples of different locations.

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The lipophilic character of phytol derivatives has been studied using reverse-phase planar chromatographic procedures. Methanol-water and acetonitrile-water binary mixtures as mobile phases with C18 and cyano as stationary phases were used in order to determine the lipophilicity parameters and C 0. The classical values were compared with the factor scores obtained by principal component analysis based also on the TLC retention data. Moderate to high correlation between the values and slopes (specific hydrophobic surface area) was observed, which reflects the involvement of secondary interactions in the TLC retention behavior. The phytol derivatives considered in this study were screened for their antituberculosis activity against the H37Rv strain. Chromatographically obtained lipophilicity parameters were correlated with the calculated log P values and minimum inhibitory concentration (MIC) values. Principal component analysis established the dominant pattern in the chromatographic indices. Quantitative structure-retention relationship and quantitative structure-activity (antitubercular) relationship investigations were performed on the lipophilicities and molecular descriptors of phytol derivatives using partial least squares. The predicted biological activities support the fact that the chromatographic processes of the investigated phytol derivatives influence the partitioning over biomembrane and are controlled mainly by the lipophilicity.

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Summary

Clerodendrum viscosum leaves are used in indigenous systems of medicines of mainland and maritime Southeast Asian countries for the treatment of fever, pain, dysentery, colic, and removal of Ascarids. The Clerodendrum species under study exhibit various phytochemical and morphological similarities. Therefore, it is very challenging to distinguish raw powdered materials used for therapeutic purposes. A validated high-performance thin-layer chromatography (HPTLC) method with 4 key markers, viz., 24ß-ethylcho-lesta-5,22E,25-triene-3ß-O-D-glucoside, clerodinin-A, 24ß-ethyl-cholesta-5,22E,25-triene-3ß-ol, and lupeol coupled with a chemometric analysis was used to distinguish 3 closely related Clerodendrum species, viz., C. inerme, C. multiforum, and C. viscosum. PRISMA approach was applied for effective HPTLC fingerprint development. The HPTLC-densitometry method was validated following the current International Conference on Harmonisation (ICH) guidelines. Taxonomic differentiation was established by fingerprint-based similarity analysis, a chemotaxonomic study using hierarchical clustering analysis (HCA), and principal component analysis (PCA) was done. HPTLC chromatogram similarity was calculated as correlation coefficient and congruence coefficient values, demonstrating poor similarities (0.26–0.86). However, PCA has resulted in 2 principal component (PC) loadings. PC1 separated C. multiforum, explaining 85.48% of variance mainly due to distribution of 2 triterpenoids. The present HPTLC method is coupled to marker-based quality determination of raw plants as well as discrimination of Clerodendrum species. Chemometric analysis based on 4 metabolites clearly establishes a practical identification of Clerodendrum species intended for therapeutic use.

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