Quantitative determination of seven chemical constituents and chemo-type differentiation of chamomiles using high-performance thin-layer chromatography

Abstract

A simple and rapid high-performance thin-layer chromatographic method was developed for the separation and determination of six flavonoids (rutin, luteolin-7-O-β-glucoside, chamaemeloside, apigenin-7-O-β-glucoside, luteolin, apigenin) and one coumarin, umbelliferone from chamomile plant samples and dietary supplements. The separation was achieved on amino silica stationary phase using dichloromethane/acetonitrile/ethyl formate/glacial acetic acid/formic acid (11:2.5:3:1.25:1.25 v/v/v/v/v) as the mobile phase. The quantitation of each compound was carried out using densitometric reflection/absorption mode at their respective absorbance maxima after postchromatographic derivatization using natural products reagent (1% w/v methanolic solution of diphenylboric acid-β-ethylamino ester). The method was validated for specificity, limits of detection and quantification, precision (intra- and interday) and accuracy. The limits of detection and quantification were found to be in the range from 6-18 and 16-55 ng/band for six flavonoids and one coumarin, respectively. The intra- and interday precision was found to be <5% RSD and recovery of all the compounds was >90%. The data acquired from high-performance thin-layer chromatography was processed by principal component analysis using XLSTAT statistical software. Application of principal component analysis and agglomerative hierarchial clustering was successfully able to differentiate two chamomiles (German and Roman) and Chrysanthemum.

Rapid discrimination of different Apiaceae species based on HPTLC fingerprints and targeted flavonoids determination using multivariate image analysis

“…Great amounts of information [variables, such as retention factor ( R f value) or features, such as colour or intensity] for a large number of samples (test objects) obtained from a thin‐layer chromatogram, require the use of statistical procedures in order to efficiently extract the maximum of useful information from the acquired data. Based on the similarity/dissimilarity analysis or correlation matrix, a number of unsupervised and supervised chemometric methods can be performed with the data, such as principal component analysis (PCA), hierarchical cluster analysis (HCA), linear discriminant analysis (LDA), partial least square discriminant analysis (PLS‐DA), k‐nearest neighbours (KNN), artificial neural networks (ANN), and partial least square (PLS) regression . This approach provides a systemic and objective way of analysing the HPTLC plate .…”

“…Based on the similarity/dissimilarity analysis or correlation matrix, a number of unsupervised and supervised chemometric methods can be performed with the data, 26 such as principal component analysis (PCA), hierarchical cluster analysis (HCA), linear discriminant analysis (LDA), partial least square discriminant analysis (PLS-DA), k-nearest neighbours (KNN), artificial neural networks (ANN), and partial least square (PLS) regression. 13,[27][28][29][30] This approach provides a systemic and objective way of analysing the HPTLC plate. 31 One of the major advantages of adopting multivariate analysis for chromatographic fingerprints is that it scrutinises the subtle differences within the chromatogram.…”

Antinociceptive activity of extracts and secondary metabolites from wild growing and micropropagated plants of Renealmia alpinia

“…HPTLC methodology is reliable, fast and cost efficient protocol that can be used to do qualitative and quantitative analysis of secondary metabolites extracted from natural sources (Lebot et al, 2014; Sagi et al, 2014). In Fig.…”

Development of a method using high‐performance liquid chromatographic fingerprint and multi‐ingredients quantitative analysis for the quality control of Yangxinshi Pian

“…Multiple‐ingredient quantitative analysis would address this problem. Multi‐ingredients quantitative analysis integrated with fingerprint analysis method has been increasingly used for the QC of Chinese herbal medicine and their preparations in recent years.…”