Single‐step screening and isolation of potential lipoxidase inhibitors from Trifolium repens by stepwise flow rate high‐speed countercurrent chromatography and semipreparative high‐performance liquid chromatography target‐guided by ultrafiltration‐LC‐MS

Abstract

An efficient method based on ultrafiltration high‐performance liquid chromatography coupled with photodiode array detector and electrospray ionization mass spectrometry for the rapid screening and identified of the ligands for activated from the extract of Trifolium repens L. Five major compounds, namely ononin, daidzein, genistein, formononetin, and biochanin A, were identified as potentially effective inhibitors. Subsequently, the specific binding ligands were separated by stepwise flow rate high‐speed countercurrent chromatography and semipreparative high‐performance liquid chromatography. This is the first report that T. repens extracts contain potent lipoxidase inhibitors. In summary, we systematically studied the active components in T. repens, evaluated their activity, separated and purified them, and identified their structure. This method is simple, fast, and efficient. It is suitable for the separation and purification of active compounds in T. repens, and provides a theoretical basis and technical platform for the development of natural medicines.

Efficient and fast screening and separation based on computer‐aided screening and complex chromatography methods for lipoxygenase inhibitors from Ganoderma lucidum

“…23 HSCCC and semi-preparative HPLC are widely used in the modern separation field due to their advantages of high automation and high compound purity. [24][25][26] Compared to conventional liquid-solid separation methods, HSCCC is a unique liquid-liquid partition chromatography method in which a solid support is not used. 27,28 This strategy eliminates the irreversible adsorptive loss of the samples onto the solid support matrix used in a conventional chromatographic column.…”

“…For certain mixtures with complex compositions, especially for the separation of triterpenoids, commonly used techniques include column chromatography, semi‐preparative high‐performance liquid chromatography (semi‐preparative HPLC), and high‐speed countercurrent chromatography (HSCCC), among others 23 . HSCCC and semi‐preparative HPLC are widely used in the modern separation field due to their advantages of high automation and high compound purity 24–26 . Compared to conventional liquid–solid separation methods, HSCCC is a unique liquid–liquid partition chromatography method in which a solid support is not used 27,28 .…”

Targeted Separation of COX-2 Inhibitor from Pterocephalus hookeri Using Preparative High-Performance Liquid Chromatography Directed by the Affinity Solid-Phase Extraction HPLC System

“…The general mode of plant-derived compound discovery with COX-2 inhibitory activity generally requires multiple-step separation and activity determination of every separated molecule, which is time consuming, labor-intensive and leads to low efficiency [ 24 ]. To enhance the discovery throughput of COX-2 inhibitors, plenty of methods are based on biological binding affinities between ligands and enzymes, for directly screening the potential COX-2 inhibitors from a complex mixture are developed.…”