By use of near infrared (NIR) spectroscopy hyphenated to thinlayer chromatography (TLC) for analysis of methoxylated flavones in a phytomedicine we sought to achieve two objectives: first, to establish a method for rapid, qualitative identification of five methoxylated flavones, denoted G1, G2, G3, G4, and G5, in order of their RF values in normal-phase TLC, and, second, to produce a quantitative model for analysis of G4 (3′,4′,5′-trimethoxyflavone), the compound most representative of Primula veris flowers in phytomedicine. To provide appropriate reference analytical data for building the multivariate cluster and partial least-squares regression (PLS) model, TLC was performed on alumina with n-hexane-ethyl acetate 70:30 (v/v) as mobile phase. Forty-four spectra of eleven independent phytomedicine samples were analyzed with five scans to generate a qualitative cluster model based on PCA (principle-components analysis) that enabled differentiation between G1-G5 on the basis of their methoxylation pattern. This PLS model, in the calibration range between 0 and 1000 mg L−1, enabled quantification of G4 with a standard error of cross validation (SECV) ,54.61 mg L−1. The possibility of conducting qualitative and quantitative analysis simultaneously by use of this method revealed NIRS to be an efficient alternative to conventional modes of detection used for analysis of G1-G5, especially in phytomedicines.
J.B. Harborne, The Flavonoids: Advances in Research Since 1986, Chapman & Hall/CRC, Boca Raton, 1994.
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E. de Rijke, P. Out, W.M.A. Niessen, F. Ariese, C. Gooijer, U.A.Th. Brinkman, J. Chromatogr. A 1112 (2006) 31–63.