Purification of a Flavonoid Sample by Countercurrent Chromatography

Abstract

A quercitrin rich vegetal flavonoid sample was fractionated using a new hydrodynamic CCC apparatus based on the It0 scheme IV design. The Kromaton CCC apparatus has two spools with a tube coiling allowing work with two different apparatus volumes. The small "analytical" volume was 94 mL and the large "preparative" volume was 1.07 L. The fractionation of the flavonoid sample was done using the two machine volumes. The bi phasic liquid system was a non chlorinated solvent system, butanonelwater. W detection was possible at 330 nm. In one run, 330 mg of the flavonoid sample could be fractionated by the CCC machine in six peaks that were collected in nine fractions. 3003 Copyright 0 1998 by Marcel Dekker, Inc. www.dekker.com Downloaded by [Erciyes University] at 22:57 02 January 2015 3004 BERTHOD ET AL.The HPLC and MS analysis of the fractions allowed identification of the main constituents of the sample. Quercitrin, a rhamno-flavonoid, was the major compound making up more than 88% wlw of the studied sample. The three minor identified flavonoids were quercetin, kaempferol, and kaempferol-3-rhamnoglucoside. Six very minor impurities were detected by the HPLC analysis of the fractions, but they were not unambiguously identified.

Strategies of solvent system selection for the isolation of flavonoids by countercurrent chromatography

“…A total of 170 papers (including review papers but excluding book chapters and patents) were found, of which we had access to 148 papers. These papers 12–159 comprise studies on two large groups of flavonoids: the free and the glycosylated flavonoids. From the first group, we found examples of the isolation of flavones, flavonols, isoflavonoids, flavanones, flavanonols, chalcones, anthocyanidines, proanthocyanidines, rotenoids, biflavonoids, triflavonoids, and theaflavins, which account for 45 references and two review papers (32% of the total).…”

Separation and Identification of Flavonoids in an Extract from the Seeds of Oroxylum indicum by CCC

“…It has also been found to be a useful purgative. [7][8][9][10] In this paper, high-speed analytical countercurrent chromatography was used to select a suitable solvent system and HSCCC was applied to the purification of the three flavonoids, followed by the identification of these using LC-MS electrospray ionisation and NMR. In the seeds of O. indicum, there are a large amount of bioactive flavonoids such as baicalein, baicalin, chrysin, apigenin.…”

Polyphenols in countercurrent chromatography. An example of large scale separation

“…Figure 2 shows the CCC chromatogram of a 0.33 g injection. The compounds were identified using mass spectrometry as described in a recent article [14]. Table IV lists the chromatographic parameters that can be established from the CCC chromatogram.…”