Development and validation of an assay for the quantification of glycosides using high-performance thin-layer chromatography (HPTLC)

Abstract

This paper presents a simple and rapid approach to the quantification of various glycosides using high-performance thin-layer chromatography (HPTLC). Different classes of glycosides, represented by genistin and ononin (both monosaccharidic O-glycosides), rutin (a disaccharidic O-glycoside) and luteolin-6-C-glucoside (a monosaccharidic C-glycoside), were successfully separated using a mixture of ethyl acetate‒methanol‒glacial acetic acid‒formic acid (11:1:1:1, V/V) as the mobile phase followed by derivatisation with natural product–polyethylene glycol (NP–PEG) reagent. The method was validated for the quantification of these glycosides in accordance with the guidelines of the International Council for Harmonisation. The general applicability of the validated approach is demonstrated with the analysis of a large number of glycosides including two glycosides (i.e. rutin, naringin) in commercial products.

Estrogenic Isoflavones in Clover Plants, Flower Nectar, Unripe Honeys and Mature Honeys: A Natural Biochemical Transformation of Isoflavones by Honeybees

“…VSA was prepared by dissolving 1 g of vanillin and 2 mL of sulfuric acid in 100 mL of methanol. NP was prepared by dissolving 1 g of 2-aminoethyl diphenylborinate in methanol before the volume of the solution was made up to 100 mL (1% m / v ) [ 25 ]. PEG reagent was prepared by mixing 5 g of polyethylene glycol in ethanol and then the volume of the solution was adjusted to 100 mL (5% m / v ) [ 25 ].…”

“…The identification and quantification of isoflavone aglycones (i.e., biochanin A, formononetin, genistein and daidzein) in different clover samples was carried out using mobile phase A [ 26 ], while for glucosylated isoflavones (i.e., sissotrin, ononin, genistin and daidzin) mobile phase B was used [ 25 ].…”