title: Characterization and identification of isoflavonoid glycosides in the root of Spiny restharrow (Ononis spinosa L.) by HPLC-QTOF-MS, HPLC-MS/MS and NMR.
abstract: Restharrow root has been used in traditional medicine for thousands of years; however, the active ingredients responsible for the diuretic effect are still unknown. Previous studies have proved that the root extract contains isoflavonoids, however only few derivatives were identified, mostly relying on retention times or UV data. The aim of our work was to perform a detailed structural characterization of the complete isoflavonoid profile in the aqueous-methanolic extract of Ononis spinosa root by high-performance liquid chromatography coupled with electrospray ionization accurate-mass quadrupole time-of-flight and tandem mass spectrometry in positive ionization mode (HPLC-ESI-QTOF-MS, HPLC-ESI-MS/MS) and nuclear magnetic resonance spectroscopy (NMR). On the basis of the accurate masses and fragmentation patterns isoflavones (formononetin, calycosin and pseudobaptigenin) and pterocarpans (maackiain and medicarpin) were identified. Two further dihydroisoflavone aglycones, namely onogenin and sativanone and a unique glucoside were isolated and their structures were elucidated by NMR experiments. Calycosin, onogenin and sativanone were detected in this plant for the first time. In contrast to previous works, the presence of biochanin A could not be confirmed, however its regioisomer calycosin and its derivatives were identified. Similarly, neither tectorigenin derivatives could be detected, however the isobar compound sativanone and its various glucosides were elucidated. The presence of genistein and daidzein could not be confirmed in the extract. Fragmentation pathways for onogenin and sativanone are presented. In the aqueous-methanolic extract 9 glucosides, 6 minor and 8 major glucoside malonates, 4 glucoside acetates and 7 aglycones were found. In total, 34 compounds were successfully identified.
authors.affiliation: Semmelweis University, Department of Pharmacognosy, Üllői út 26, H-1085 Budapest, Hungary.
authors.forename: Nóra
authors.initials: N
authors.lastname: Gampe
authors.affiliation: Semmelweis University, Department of Pharmacognosy, Üllői út 26, H-1085 Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Darcsi
authors.affiliation: National Institute of Pharmacy and Nutrition, Zrínyi u. 3, H-1051 Budapest, Hungary.
authors.forename: Szilvia
authors.initials: S
authors.lastname: Lohner
authors.affiliation: Semmelweis University, Department of Pharmacognosy, Üllői út 26, H-1085 Budapest, Hungary. Electronic address: beni.szabolcs@pharma.semmelweis-univ.hu.
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Béni
authors.affiliation: Semmelweis University, Department of Pharmacognosy, Üllői út 26, H-1085 Budapest, Hungary. Electronic address: kursinszki.laszlo@pharma.semmelweis-univ.hu.
authors.forename: László
authors.initials: L
authors.lastname: Kursinszki
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019833:Genistein / Q000737:chemistry; D005960:Glucosides / Q000737:chemistry; D006027:Glycosides / Q000737:chemistry*; D007529:Isoflavones / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D000068757:Ononis / Q000737:chemistry*; D018517:Plant Roots / Q000737:chemistry*; D036343:Pterocarpans / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods; D000094162:Phytoalexins
chemical_list: D019833:Genistein; D005960:Glucosides; D006027:Glycosides; D007529:Isoflavones; D036343:Pterocarpans; C000607319:sativanone; C121707:7,3'-dihydroxy-4'-methoxyisoflavone; C007768:formononetin; C004742:daidzein; C047353:medicarpin; D000094162:Phytoalexins; C120039:tectorigenin; C060800:psi-baptigenin; C001449:inermin
keywords: Dihydroisoflavone; Isoflavone; MS fragmentation pattern; Pterocarpan; Structural analysis