title: Ultrafiltration-LC-MS combined with semi-preparative HPLC for the simultaneous screening and isolation of lactate dehydrogenase inhibitors from Belamcanda chinensis.
abstract: Stroke represents the fourth leading cause of death in the USA and the second leading cause of death worldwide. Lactate dehydrogenase inhibitors are widely used in the treatment of ischemic stroke and natural products are considered a promising source of novel lactate dehydrogenase inhibitors. In this study, we used PC12 cells to determine the protective effect of extracts from the herb Belamcanda chinensis following toxic challenge. Using ultrafiltration high-performance liquid chromatography coupled with photo-diode array detection and electrospray ionization mass spectrometry, we screened and identified isoflavonoids from Belamcanda chinensis extracts. Semi-preparative high-performance liquid chromatography was then applied to separate and isolate the active constituents. Using these methods, we identified six major compounds in Belamcanda chinensis as lactate dehydrogenase inhibitors: tectoridin, iristectorin A, iridin, tectorigenin, irigenin, and irisflorentin, which were then isolated to >92% purity. This is the first report that Belamcanda chinensis extracts contain potent lactate dehydrogenase inhibitors. Our results demonstrate that the systematic isolation of bioactive components from Belamcanda chinensis guided by ultrafiltration high-performance liquid chromatography coupled with photo-diode array detection and electrospray ionization mass spectrometry represents a feasible and efficient technique that could be extended for the identification and isolation of other enzyme inhibitors.
authors.affiliation: Central Laboratory, Changchun Normal University, Changchun, China.
authors.forename: Senlin
authors.initials: S
authors.lastname: Li
authors.affiliation: Central Laboratory, Changchun Normal University, Changchun, China.
authors.forename: Sainan
authors.initials: S
authors.lastname: Li
authors.affiliation: Central Laboratory, Changchun Normal University, Changchun, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Central Laboratory, Changchun Normal University, Changchun, China.
authors.forename: Chunming
authors.initials: C
authors.lastname: Liu
authors.affiliation: Central Laboratory, Changchun Normal University, Changchun, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Chen
authors.affiliation: Central Laboratory, Changchun Normal University, Changchun, China.
authors.forename: Yuchi
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002545:Brain Ischemia; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004791:Enzyme Inhibitors / Q000302:isolation & purification*; D027621:Iridaceae / Q000737:chemistry*; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors*; D016716:PC12 Cells; D064209:Phytochemicals / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization; D020521:Stroke; D053719:Tandem Mass Spectrometry; D014462:Ultrafiltration
chemical_list: D004791:Enzyme Inhibitors; D064209:Phytochemicals; D010936:Plant Extracts; D007770:L-Lactate Dehydrogenase
keywords: Inhibitors; Ischemic stroke; PC12 cells; Semi-preparative high-performance liquid chromatography; Traditional Chinese medicine