Qualitative and Quantitative Phytochemical Analysis of Ononis Hairy Root Cultures

Abstract

Hairy root cultures are genetically and biochemically stable, and they regularly possess the same or better biosynthetic capabilities for specialized (secondary) metabolite production compared to the intact plant. Ononis species are well-known herbal remedies in ethnopharmacology and rich sources of isoflavonoids. Besides isoflavones, less prevalent isoflavones and pterocarpans with valuable biological effects can be found in Ononis species as well. As these plants are only collected but not cultivated, biotechnological methods could play a role in the larger-scale extraction of Ononis isoflavonoids. Regarding this information, we aimed to establish Ononis spinosa and Ononis arvensis hairy root cultures (HRCs) and analyze the isoflavonoid profile of hairy root cultures qualitatively and quantitatively, in order to define their capacity to produce biologically valuable isoflavonoids. During the qualitative description, beside isoflavonoids, two new phenolic lactones, namely, bulatlactone 2″-O-β-D-glucoside and ononilactone, were isolated, and their structures were characterized for the first time. Altogether, 29 compounds were identified by the means of UPLC-Orbitrap-MS/MS. Based on UHPLC-UV-DAD measurements, the isoflavonoid spectrum of the Ononis HRCs differed markedly from wild-grown samples, as they produce a limited range of the scaffolds. The most abundant compounds in the HRCs were medicarpin glucoside and sativanone glucoside. The overall isoflavonoid production of the cultures was comparable to wild-grown O. arvensis and approximately twice as high as in wild-grown O. spinosa samples. As the overall content of wild-grown samples include more isoflavonoid derivatives, the HRCs contain structurally less divergent isoflavonoids but in higher quantity.

Hairy Root Cultures as a Source of Polyphenolic Antioxidants: Flavonoids, Stilbenoids and Hydrolyzable Tannins

“…The transformed root cultures of Ononis spinosa L. and Ononis arvensis L. (Fabaceae) were obtained by Nóra Gampe et al [ 170 ]. The most abundant compounds in the HRs were medicarpin, sativanone and pseudobaptigenin glucosides (16.9–28.9 mg/g DW, 1.3–11.4 mg/g DW and 0.9–2.0 mg/g DW, respectively).…”

Identification and Pharmacological Activities of Secondary Metabolites of Phytochemicals

“…Both techniques have been used to separate, identify and quantify series of structurally different SMs in plants include spectrophotometric [57,[73][74][75], GC-MS [56,76], LC-MS [77], LC-MS/MS [56,76,78], Liquid Chromatography-High Resolution Electrospray Ionization Mass Spectrometry (LC-HRESIMS) [57], Nuclear Magnetic Resonance (NMR) [44,79], Ultra-Fast Liquid Chromatography (UFLC-Q-TOF-MS/MS) [80][81], UFLC-triple-Q-TOF-MS/MS [82], Ultra-high performance Liquid Chromatography quadrupole-time of flight mass spectrometry [83], UHPLC-LTQ-Orbitrap [84], UHPLC/Q-Orbitrap-MS [85], HPLC-DAD [86], UHPLC-Q-TOF-MS [87], UHPLC-Q-TOF-MS/MS [88], HPLC-DAD-ESI-MS [89], UPLC-QTOF/MSE [90], UHPLC-MS/MS [91], HPLC-ESI-QTOF-MS [92], UPLC-QTOF-MS/MS [93], UHPLC-Q-TOF-MS/MS [94][95], UHPLC-ESI-MS/MS [96], UHPLC-ESI-Orbitrap-MS/MS [97], UPLC-IT-MS [98], UPLC-Q-TOF-MS [98], UPLC-PDA [45], HPLC-ESI-MS and MS 2 [99], reverse phase high performance liquid chromatography-diode array detector (RP-HPLC-DAD) [74], HPLC-HRMS [100]. been used to produce a yellow color that turn colorless on addition of dilute hydrochloric acid confirm flavonoid [102,[104][105]…”