Preparative separation of flavonoids in plant extract of Smilacis Glabrae Roxb. by high performance counter-current chromatography

Abstract

Four flavonoids, isoastilbin, astilbin, isoengelitin, and engelitin were isolated and purified simultaneously from Smilacis Glabrae Roxb. for the first time by high performance counter-current chromatography using a system consisting of n-hexane-n-butanol-water (1:2:3, v/v/v). A total of 392.6 mg of astilbin, 71.4 mg of isoastilbin, 47.4 mg of engelitin, and 10.3 mg of isoengelitin were purified from 1.89 g of the ethyl acetate extract of Smilacis Glabrae Roxb. in six runs, each at over 94.51% purity as determined by HPLC. The structures of the four compounds were identified by their retention time, the LC-ESI-MS(n) in the negative ion mode, and confirmed by (1)H-NMR experiments. The characteristic LC-ESI-MS fragmentation patterns of the four compounds were discussed.

Simultaneous chemical fingerprint and quantitative analysis of Rhizoma Smilacis Glabrae by accelerated solvent extraction and high‐performance liquid chromatography with tandem mass spectrometry

“…The three substances were introduced into the mass spectrometer and the chemical structures of the three flavonoids as standards were also confirmed by 1 H and 13 C NMR spectroscopy. The ESI‐MS and 1 H and 13 C NMR spectroscopy data were consistent with those in the literature .…”

Solvent system selectivities in countercurrent chromatography using Salicornia gaudichaudiana metabolites as practical example with off-line electrospray mass-spectrometry injection profiling

“…It can be considered as a combination of two binary solvent systems: n-hexane-methanol and ethyl acetate-water [5,40]. (ii) The second solvent system family composed of nhexane-butanol-water (HBuWat) had been used for the isolation of glycosylated flavonoids and less polar saponins [45][46][47][48][49][50]. The HBuWat solvent system is composed of two miscible solvents plus a third solvent that is immiscible with the previous two, being called type 2 solvent system [5].…”

Separation of Seven Polyphenols from the Rhizome of Smilax glabra by Offline Two Dimension Recycling HSCCC with Extrusion Mode

“…With advantages such as high sample recovery, large loading capacity, low risk of sample denaturation, and irreversible adsorption [ 9 ], conventional high-speed countercurrent chromatography (HSCCC) has been applied for the separation of polyphenols from the rhizome of S. glabra . However, because the chemical structures of the polyphenols were similar, (four of them were isomers, and the partition coefficients ( K -value) of the polyphenols had small differences), only two purified compounds (astilbin and isoastilbin) of these isomers were obtained [ 10 , 11 ]. Thus, effective methods for separation of polyphenols from the rhizome of S. glabra need to be developed.…”