Cutting-edge analytical technologies for the comprehensive metabolic profiling of Alkanna tinctoria roots cultured in greenhouse conditions

Rodić, N;Vlachou, A;Papakonstantinou, C;Zhao, Y;Assimopoulou, AN;et.al.
(2019) International ongress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research — Location: Innsbruck, Austria (1.September.2019)

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Cutting-edgeanalyticaltechnologiesforthecomprehensivemetabolicprofilingofAlkannatinctoriarootsculturedingreenhouse.docx
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Authors
  • Rodić, N
    Author
  • Vlachou, A
    Author
  • Papakonstantinou, C
    Author
  • Zhao, YUCLouvain
    Author
  • Cartabia, AUCLouvain
    Author
  • Lalaymia, IUCLouvain
    Author
  • Declerck, SUCLouvain
    Author
  • Assimopoulou, AN
    Author
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Abstract
The use of plants containing naphthoquinone derivatives Alkannins & Shikonins (A/S) by humans dates back to ancient times. In recent decades, the use of A/S has seen a resurgence and A/S have risen to a pivotal role as pharmaceutical and cosmeceutical agents, since they possess strong wound healing, antimicrobial, anti-inflammatory, tissue regenerative and antitumor properties. It is thus crucial to enhance the biosynthesis of bioactive A/S in Alkanna tinctoria plants, that naturally produce high amounts of these metabolites [1]. In the frame of “MICROMETABOLITE” EU H2020 project, we have optimized a workflow for the metabolic profiling of A. tinctoria roots, cultured in the greenhouse from plants obtained by in vivo shoot cuttings. A fast and reliable extraction procedure was achieved for comprehensive profiling and identification of A/S and other metabolites biosynthesized in the roots. The aim of this work was to determine the growth stage with peak A/S production, while simultaneously obtaining additional information on the root metabolome. A combination of UHPLC-HRMS and NMR was used for metabolite identification, HPLC was utilized for reliable quantitation of A/S and the extracts were subjected to chiral HPLC analysis [2] for determination of the enantiomeric A/S ratio. Different A/S derivatives and other metabolites were identified in plant roots using UHPLC-HRMS and NMR. Six A/S derivatives and total A/S were quantified using HPLC-DAD. From six vegetation stages of A. tinctoria grown under greenhouse conditions, fruiting period was found to peak A/S production (1% wt/wt of root), while the enantiomeric alkannin/shikonin ratio remained constant (93.7%).
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Rodić, N., Vlachou, A., Papakonstantinou, C., Zhao, Y., Cartabia, A., Lalaymia, I., Grigoriadou, K., Krigas, N., Maloupa, E., Declerck, S., Gika, H., Papageorgiou, V., & Assimopoulou, A. (2019). Cutting-edge analytical technologies for the comprehensive metabolic profiling of Alkanna tinctoria roots cultured in greenhouse conditions. International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research. Published. International ongress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research, Innsbruck, Austria. https://doi.org/10.1055/s-0039-3399826 (Original work published 2019)