One-step separation of nine structural analogues from Poria cocos (Schw.) Wolf. via tandem high-speed counter-current chromatography.

A novel one-step separation strategy-tandem high-speed counter-current chromatography (HSCCC) was developed with a six-port valve serving as the switch interface. Nine structural analogues including three isomers were successfully isolated from Poria cocos (Schw.) Wolf. by one step. Compared with conventional HSCCC, peak resolution of target compounds was effectively improved in tandem one. Purities of isolated compounds were all over 90% as determined by HPLC. Their structures were then identified via UV, MS and (1)H NMR, and eventually assigned as poricoic acid B (1), poricoic acid A (2), 3ß,16α-dihydroxylanosta-7, 9(11), 24-trien-21-oic acid (3), dehydrotumulosic acid (4), polyporenic acid C (5), 3-epi-dehydrotumulosic acid (6), 3-o-acetyl-16α-hydroxydehydrotrametenolic acid (7), dehydropachymic acid (8) and dehydrotrametenolic acid (9) respectively. The results indicated that tandem HSCCC can effectively improve peak resolution of target compounds, and can be a good candidate for HSCCC separation of structural analogues.

Separation of α-amylase inhibitors from Abelmoschus esculentus (L).Moench by on-line two-dimensional high-speed counter-current chromatography target-guided by ultrafiltration-HPLC.

In this study, an on-line two-dimensional high-speed counter-current chromatography system based on a six-port valve was developed. Target-guided by ultrafiltration with high-performance liquid chromatography, the one-step isolation of three potential α-amylase inhibitors from Abelmoschus esculentus (L).Moench was achieved by employing the developed orthogonal system and extrusion elution mode. The purities of three potential α-amylase inhibitors were all over 95% as determined by high-performance liquid chromatography. Furthermore, UV, mass spectrometry and 1 H NMR spectroscopy were applied to the structural identification of the isolated three target compounds, their structures were assigned as quercetin-3-O-sophoroside (i), 5,7,3',4'-tetrahydroxy flavonol-3-O-[ß-d-rhamnopyranosil-(1→2)]-ß-d-glucopyranoside (ii ) and isoquercitrin (iii), respectively. The Results demonstrated that the proposed method was highly efficient to screen and isolate enzyme inhibitors from complex natural products extracts, and on-line two-dimensional high-speed counter-current chromatography can effectively increase the peak resolution of target compounds.

Separation of polyphenols from leaves of Malus hupehensis (Pamp.) Rehder by off-line two-dimensional High Speed Counter-Current Chromatography combined with recycling elution mode.

In this study, off-line two-dimensional High Speed Counter-Current Chromatography (2D HSCCC) strategy combined with recycling elution mode was developed to isolate compounds from the ethyl acetate extract of a common green tea--leaves of Malus hupehensis (Pamp.) Rehder. In the orthogonal separation system, a conventional HSCCC was employed for the first dimension and two recycling HSCCCs were used for the second in parallel. Using a solvent system consisting of n-hexane-ethyl acetate-methanol-water (1:4:0.6:4.4, v/v) in the first and second dimension, four compounds including 3-hydroxy-phlorizin (1), phloretin (2), avicularin (3) and kaempferol 3-O-ß-D-glucoside (4) were obtained. The purities of these four compounds were all over 95.0% as determined by HPLC. And their structures were all identified through UV, MS and (1)H NMR. It has been demonstrated that the combination of off-line 2D HSCCC with recycling elution mode is an efficient technique to isolate compounds with similar polarities in natural products.

Separation of five flavonoids from tartary buckwheat (Fagopyrum tataricum (L.) Gaertn) grains via off-line two dimensional high-speed counter-current chromatography.

An off-line two dimensional (2D) high-speed counter-current chromatography (HSCCC) strategy was successfully used for preparative separation of five flavonoids from tartary buckwheat (Fagopyrum tataricum (L.) Gaertn) grains with different solvent systems for the first time in this paper. n-Hexane-ethyl acetate-methanol-water 3:5:3:5 (v/v) was selected as the first dimension solvent system to purify quercetin (4) and kaempferol (5). The second dimension solvent system, ethyl acetate-n-butanol-water 7:3:10 (v/v), was used to isolate quercetin 3-O-rutinoside-3'-O-ß-glucopyranoside (1), rutin (2) and kaempferol 3-rutinoside (3). The purities of these compounds were all above 96.0% and their structures were identified through UV, MS and (1)H NMR. The results indicated that the off-line 2D HSCCC is an efficient technique to isolate flavonoids compounds from grains.

Separation of five compounds from leaves of Andrographis paniculata (Burm. f.) Nees by off-line two-dimensional high-speed counter-current chromatography combined with gradient and recycling elution.

An off-line two-dimensional high-speed counter-current chromatography method combined with gradient and recycling elution mode was established to isolate terpenoids and flavones from the leaves of Andrographis paniculata (Burm. f.) Nees. By using the solvent systems composed of n-hexane/ethyl acetate/methanol/water with different volume ratios, five compounds including roseooside, 5,4'-dihydroxyflavonoid-7-O-ß-d-pyranglucuronatebutylester, 7,8-dimethoxy-2'-hydroxy-5-O-ß-d-glucopyranosyloxyflavon, 14-deoxyandrographiside, and andrographolide were successfully isolated. Purities of these isolated compounds were all over 95% as determined by high-performance liquid chromatography. Their structures were identified by UV, mass spectrometry, and (1) H NMR spectroscopy. It has been demonstrated that the combination of off-line two-dimensional high-speed counter-current chromatography with different elution modes is an efficient technique to isolate compounds from complex natural product extracts.