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.