Chemoprofiling of Himantoglossum robertianum (Loisel.) P. Delforge leaves reveals predominance of gastrodigenin and structurally related compounds

doi:10.1007/s13659-025-00526-7

Abstract

Abstract: PurposeThe purpose of this study was to phytochemically profile Himantoglossum robertianum leaves. In fact, despite its wide distribution and its use in traditional medicine, this orchid is still understudied and little is known about its phytochemicals.MethodsThe analyses were performed by ¹H NMR fingerprinting, elucidated by further 2D NMR and UHPLC-MS experiments. Both primary and secondary metabolites were qualified and quantified. The study was carried out comparing six natural populations by metabolomics approach, allowing further considerations on the influence of the environment on the concentration of metabolites.ResultsThis work brings to light a surprising phytochemical parallel between H. robertianum and the medicinal orchid Gastrodia elata. In fact, the most abundant specialized metabolites resulted: gastrodigenin, gastrodin, bis(4-hydroxybenzyl)ether, parishin A, parishin C, and parishin E. Interestingly, these metabolites are all known for their potential in the treatment of neurological disorders and are, indeed, the active principles of Gastrodia elata, an important orchid used in Traditional Chinese Medicine. The active metabolites were present in all the natural populations, where only slight variations in their concentration were revealed.ConclusionMapping the metabolome of H. robertianum leaves has provided new insights into the study of orchids, including diagnostic signals for rapid identification of gastrodigenin-like compounds directly from the ¹H NMR profile of a crude extract. From a bioprospecting perspective, finding active metabolites in leaves makes the plant source more valuable than the perennial hypogeal organs that are usually the herbal drug of orchids (i.e. G. elata).

Key words: Himantoglossum robertianum, NMR metabolomics, Orchids, Gastrodia elata, Parishin, 4-hydroxybenzyl alcohol

摘要： PurposeThe purpose of this study was to phytochemically profile Himantoglossum robertianum leaves. In fact, despite its wide distribution and its use in traditional medicine, this orchid is still understudied and little is known about its phytochemicals.MethodsThe analyses were performed by ¹H NMR fingerprinting, elucidated by further 2D NMR and UHPLC-MS experiments. Both primary and secondary metabolites were qualified and quantified. The study was carried out comparing six natural populations by metabolomics approach, allowing further considerations on the influence of the environment on the concentration of metabolites.ResultsThis work brings to light a surprising phytochemical parallel between H. robertianum and the medicinal orchid Gastrodia elata. In fact, the most abundant specialized metabolites resulted: gastrodigenin, gastrodin, bis(4-hydroxybenzyl)ether, parishin A, parishin C, and parishin E. Interestingly, these metabolites are all known for their potential in the treatment of neurological disorders and are, indeed, the active principles of Gastrodia elata, an important orchid used in Traditional Chinese Medicine. The active metabolites were present in all the natural populations, where only slight variations in their concentration were revealed.ConclusionMapping the metabolome of H. robertianum leaves has provided new insights into the study of orchids, including diagnostic signals for rapid identification of gastrodigenin-like compounds directly from the ¹H NMR profile of a crude extract. From a bioprospecting perspective, finding active metabolites in leaves makes the plant source more valuable than the perennial hypogeal organs that are usually the herbal drug of orchids (i.e. G. elata).

关键词: Himantoglossum robertianum, NMR metabolomics, Orchids, Gastrodia elata, Parishin, 4-hydroxybenzyl alcohol

Ilaria Chiocchio, Antonio De Agostini, Manuela Mandrone, Pierluigi Cortis, Clarissa Tarozzi, Ferruccio Poli, Cinzia Sanna. Chemoprofiling of Himantoglossum robertianum (Loisel.) P. Delforge leaves reveals predominance of gastrodigenin and structurally related compounds[J]. Natural Products and Bioprospecting, 2025, 15(5): 45-45.

References

[1] Ghai D, Verma J, Kaur A, Thakur K, Pawar SV, Sembi JK. Bioprospection of orchids and appraisal of their therapeutic indications. Bioprospect Plant Biodivers Ind Mol. 2021. https://doi.org/10.1002/9781119718017.ch20.
[2] Sut S, Maggi F, Dall’Acqua S. Bioactive secondary metabolites from orchids (Orchidaceae). Chem Biodivers. 2017. https://doi.org/10.1002/cbdv.201700172.
[3] Bateman RM, Molnár AV, Sramkó G. In situ morphometric survey elucidates the evolutionary systematics of the Eurasian Himantoglossum clade (Orchidaceae: Orchidinae). PeerJ. 2017. https://doi.org/10.7717/peerj.2893.
[4] Caliskan O, Beşir A, Anıl M, Gülser C, Yazıcı F, Kurt D. Morphologic and chemical characterizations of some salep orchids. Eurasian J Soil Sci. 2023;12:344-51. https://doi.org/10.1839/ejss.1333347.
[5] Fekete R, Nagy T, Bódis J, Biró É, Löki V, Süveges K, Takács A, Tökölyi J, Molnár AV. Roadside verges as habitats for endangered lizard-orchids (Himantoglossum spp.): ecological traps or refuges? Sci Total Environ. 2017;607-608:1001-8. https://doi.org/10.1016/j.scitotenv.2017.07.037.
[6] Sramkó G, Molnár AV, Hawkins JA, Bateman RM. Molecular phylogeny and evolutionary history of the Eurasiatic orchid genus Himantoglossum s.l. (Orchidaceae). Ann Bot. 2014;114:1609-26. https://doi.org/10.1093/aob/mcu179.
[7] Delforge P. Orchids of Europe, North Africa and the Middle East. Oregon, USA: Timber Press Inc.; 2006.
[8] Geraci A, Amato F, Di Noto G, Bazan G, Schicchi R. The wild taxa utilized as vegetables in Sicily (Italy): a traditional component of the Mediterranean diet. J Ethnobiol Ethnomed. 2018;14:1-27. https://doi.org/10.1186/s13002-018-0215-x.
[9] Paulis G. I nomi popolari delle piante in Sardegna. Etimologia, Storia, Tradizioni. Sassari, Italy: Carlo Delfino Editore; 1992.
[10] De Agostini A, Robustelli della Cuna FS, Cortis P, Cogoni A, Sottani C, Soddu F, Sanna C. Volatile organic compounds (VOCs) diversity in the orchid Himantoglossum robertianum (Loisel.) P. Delforge from Sardinia (Italy). Diversity. 2022. https://doi.org/10.3390/d14121125.
[11] Gallego E, Gelabert A, Roca FJ, Perales JF, Guardino X. Identification of volatile organic compounds (VOC) emitted from three European orchid species with different pollination strategies: two deceptive orchids (Himantoglossum robertianum and Ophrys apifera) and a rewarding orchid (Gymnadenia conopsea). J Biodivers Environ Sci. 2012;2(5):18-29.
[12] Romano VA, Rosati L, Fascetti S, Cittadini AMR, Racioppi R, Lorenz R, D’Auria M. Spatial and temporal variability of the floral scent emitted by Barlia robertiana (Loisel.) Greuter, a Mediterranean food-deceptive orchid. Compounds. 2022;2:37-53. https://doi.org/10.3390/compounds2010004.
[13] Bazzicalupo M, Burlando B, Denaro M, Barreca D, Trombetta D, Smeriglio A, Cornara L. Polyphenol characterization and skin-preserving properties of hydroalcoholic flower extract from Himantoglossum robertianum (Orchidaceae). Plants. 2019. https://doi.org/10.3390/plants8110502.
[14] Badalamenti N, Russi S, Bruno M, Maresca V, Vaglica A, Ilardi V, Zanfardino A, Di Napoli M, Varcamonti M, Cianciullo P, et al. Dihydrophenanthrenes from a Sicilian accession of Himantoglossum robertianum (Loisel.) P. Delforge showed antioxidant, antimicrobial, and antiproliferative activities. Plants. 2021. https://doi.org/10.3390/plants10122776.
[15] Foito A, Stewart D. Metabolomics: a high-throughput screen for biochemical and bioactivity diversity in plants and crops. Curr Pharm Des. 2018;24:2043-54. https://doi.org/10.2174/1381612824666180515125926.
[16] Hu Y, Li C, Shen W. Gastrodin alleviates memory deficits and reduces neuropathology in a mouse model of Alzheimer’s disease. Neuropathology. 2014;34:370-7. https://doi.org/10.1111/neup.12115.
[17] Wang Y, Zhang M, Zhou X, Xu C, Zhu C, Yuan Y, Chen N, Yang Y, Guo Q, Shi J. Insight into medicinal chemistry behind traditional Chinese medicines: p-Hydroxybenzyl alcohol-derived dimers and trimers from Gastrodia elata. Nat Prod Bioprospect. 2021;11:31-50. https://doi.org/10.1007/s13659-020-00258-w.
[18] Zhou J, Chen JQ, Gong S, Ban YJ, Zhang L, Liu Y, Wu JL, Li N. Isolation, bioactivity, and molecular docking of a rare gastrodin isocitrate and diverse parishin derivatives from Gastrodia elata Blume. ACS Omega. 2024;9:14520-9. https://doi.org/10.1021/acsomega.4c00436.
[19] Lim EJ, Jung HJ, Park EH, Kang HJ. Anti-angiogenic, anti-inflammatory and anti-nociceptive activity of 4-hydroxybenzyl alcohol. J Pharm Pharmacol. 2007;59:1235-40. https://doi.org/10.1211/jpp.59.9.0007.
[20] Yu SS, Zhao J, Lei SP, Lin XM, Wang LL, Zhao Y. 4-hydroxybenzyl alcohol ameliorates cerebral injury in rats by antioxidant action. Neurochem Res. 2011;36:339-46. https://doi.org/10.1007/s11064-010-0335-8.
[21] Yu S, Zhao J, Wang X, Lei S, Wu X, Chen Y, Wu J, Zhao Y. 4-hydroxybenzyl alcohol confers neuroprotection through up-regulation of antioxidant protein expression. Neurochem Res. 2013;38:1501-16. https://doi.org/10.1007/s11064-013-1052-x.
[22] He X, Chen X, Yang Y, Gu J, Xie Y, Liu Y, Hao M, Heinrich M. The role of gastrodin in the management of CNS-related diseases: underlying mechanisms to therapeutic perspectives. Phytother Res. 2024. https://doi.org/10.1002/ptr.8314.
[23] Ma T, Chen P, Dong H, Wang X. Identification of key anti-neuroinflammatory components in Gastrodiae Rhizoma based on spectrum-effect relationships and its mechanism exploration. J Pharm Biomed Anal. 2024;248:116266. https://doi.org/10.1016/j.jpba.2024.116266.
[24] Wang T, Chen H, Xia S, Chen X, Sun H, Xu Z. Ameliorative effect of Parishin C against cerebral ischemia-induced brain tissue injury by reducing oxidative stress and inflammatory responses in rat model. Neuropsychiatr Dis Treat. 2021. https://doi.org/10.2147/ndt.s309065.
[25] Liu Z, Wang W, Feng N, Wang L, Shi J, Wang X. Parishin C’s prevention of Aβ 1-42-induced inhibition of long-term potentiation is related to NMDA receptors. Acta Pharm Sin B. 2016;6:189-97. https://doi.org/10.1016/j.apsb.2016.03.009.
[26] Shin EJ, Whang WK, Kim S, Bach JH, Kim JM, Nguyen XK, Lan Nguyen TT, Jung BD, Yamada K, Nabeshima T, et al. Parishin C attenuates phencyclidine-induced schizophrenia-like psychosis in mice: involvements of 5-HT1A receptor. J Pharmacol Sci. 2010;113:404-8. https://doi.org/10.1254/jphs.10040sc.
[27] Zhao X, Zhou S, Liu Y, Gong C, Xiang L, Li S, Wang P, Wang Y, Sun L, Zhang Q, et al. Parishin alleviates vascular ageing in mice by upregulation of Klotho. J Cell Mol Med. 2023;27:1398-409. https://doi.org/10.1111/jcmm.17740.
[28] Pyo MK, Jin JL, Koo YK, Yun Choi HS. Phenolic and furan type compounds isolated from Gastrodia elata and their anti-platelet effects. Arch Pharm Res. 2004;27:381-5. https://doi.org/10.1007/bf02980077.
[29] Hase T, Takao H, Iwagawa T. The bitter iridoids from Viburnum urceolatum. Phytochemistry. 1983;22:1977-82. https://doi.org/10.1016/0031-9422(83)80027-2.
[30] Kobayashi T, Higashi K, Kamada H. 4-hydroxybenzyl alcohol accumulates in flowers and developing fruits of carrot and inhibits seed formation. J Plant Physiol. 2003;160:713-6. https://doi.org/10.1078/0176-1617-00967.
[31] Long H, Luo D, Yang Y, Zhang L, Pu DB, Li J, Chen XJ, Zhu X, Liu S, Gao JB, et al. Two new phenolic compounds from the seeds of Machilus yunnanensis. J Asian Nat Prod Res. 2016;18:952-8. https://doi.org/10.1080/10286020.2016.1187139.
[32] Popova IE, Morra MJ. Sinapis alba seed meal as a feedstock for extracting the natural tyrosinase inhibitor 4-hydroxybenzyl alcohol. Ind Crops Prod. 2018;124:505-9. https://doi.org/10.1016/j.indcrop.2018.07.083.
[33] Wan JF, Yuan JQ, Mei ZN, Yang XZ. Phenolic glycosides from Boschniakia himalaica. Chin Chem Lett. 2012;23:579-82. https://doi.org/10.1016/j.cclet.2012.02.001.
[34] Zidorn C, Ellmerer-Müller EP, Stuppner H. A germacranolide and three hydroxybenzyl alcohol derivatives from Hieracium murorum and Crepis bocconi. Phytochem Anal. 2001;12:281-5. https://doi.org/10.1002/pca.585.
[35] Kim DW, Kim JK, Gebru YA, Kim YH, Choi HS, Kim MK. Identification of novel parishin compounds from the twig of Maclura tricuspidata and comparative analysis of parishin derivatives in different parts. Molecules. 2022. https://doi.org/10.3390/molecules28010007.
[36] Han SW, Wang XJ, Cui BS, Sun H, Chen H, Ferreira D, Li S, Hamann MT. Hepatoprotective glucosyloxybenzyl 2-hydroxy-2-isobutylsuccinates from Pleione yunnanensis. J Nat Prod. 2021;84:738-49. https://doi.org/10.1021/acs.jnatprod.0c01117.
[37] Hada S, Yadav DK, Roat P, Kumari N. Eulophia nuda: a review of its traditional uses, phytochemistry and pharmacology. Pharm Chem J. 2020;54:40-5. https://doi.org/10.1007/s11094-020-02152-8.
[38] Yang B, Liu H, Yang J, Gupta VK, Jiang Y. New insights on bioactivities and biosynthesis of flavonoid glycosides. Trends Food Sci Technol. 2018;79:116-24. https://doi.org/10.1016/j.tifs.2018.07.006.
[39] Ranilla LG, Rios-Gonzales BA, Ramírez-Pinto MF, Fuentealba C, Pedreschi R, Shetty K. Primary and phenolic metabolites analyses, in vitro health-relevant bioactivity and physical characteristics of purple corn (Zea mays L.) grown at two andean geographical locations. Metabolites. 2021. https://doi.org/10.3390/metabo11110722.
[40] Ashihara H. Trigonelline (N-methylnicotinic acid) biosynthesis and its biological role in plants. Nat Prod Commun. 2008;3:1423-8. https://doi.org/10.1177/1934578x0800300906.
[41] Leng C, Hou M, Xing Y, Chen J. Perspective and challenges of mycorrhizal symbiosis in orchid medicinal plants. Chin Herb Med. 2024;16:172. https://doi.org/10.1016/j.chmed.2024.03.001.
[42] Calevo J, Copetta A, Marchioni I, Bazzicalupo M, Pianta M, Shirmohammadi N, Cornara L, Giovannini A. The use of a new culture medium and organic supplement to improve in vitro early stage development of five orchid species. Plant Biosyst. 2022;156:143-51. https://doi.org/10.1080/11263504.2020.1840454.
[43] Aru A, Baldaccini P, Vacca A, Delogu G, Dessena MA, Madrau S, Melis RT, Vacca S. Carta dei suoli della Sardegna, in scala 1:250.000. 1990.
[44] Aru A, Baldaccini P, Vacca A, Delogu G, Dessena MA, Madrau S, Melis RT, Vacca S. Nota illustrativa alla Carta dei Suoli della Sardegna. 1991.
[45] Taguchi H, Yosioka I, Yamasaki K, Kim IH. Studies on the constituents of Gastrodia elata Blume. Chem Pharm Bull. 1981;29:55-62. https://doi.org/10.1248/cpb.29.55.
[46] Dong H, Yao X, Liu D, Zhou T, Wu G, Zheng X, Wang X. Effect of inorganic salt on partition of high-polarity parishins in two-phase solvent systems and separation by high-speed counter-current chromatography from Gastrodia elata Blume. J Sep Sci. 2019;42:871-7. https://doi.org/10.1002/jssc.201801012.