title: Metabolic fingerprinting of Leontopodium species (Asteraceae) by means of ¹H NMR and HPLC-ESI-MS.
abstract: The genus Leontopodium, mainly distributed in Central and Eastern Asia, consists of ca. 34-58 different species. The European Leontopodium alpinum, commonly known as Edelweiss, has a long tradition in folk medicine. Recent research has resulted in the identification of prior unknown secondary metabolites, some of them with interesting biological activities. Despite this, nearly nothing is known about the Asian species of the genus. In this study, we applied proton nuclear magnetic resonance (¹H NMR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) metabolic fingerprinting to reveal insights into the metabolic patterns of 11 different Leontopodium species, and to conclude on their taxonomic relationship. Principal component analysis (PCA) of ¹H NMR fingerprints revealed two species groups. Discriminators for these groups were identified as fatty acids and sucrose for group A, and ent-kaurenoic acid and derivatives thereof for group B. Five diterpenes together with one sesquiterpene were isolated from Leontopodium franchetii roots; the compounds were described for the first time for L. franchetii: ent-kaur-16-en-19-oic acid, methyl-15α-angeloyloxy-ent-kaur-16-en-19-oate, methyl-ent-kaur-16-en-19-oate, 8-acetoxymodhephene, 19-acetoxy-ent-kaur-16-ene, methyl-15β-angeloyloxy-16,17-epoxy-ent-kauran-19-oate. In addition, differences in the metabolic profile between collected and cultivated species could be observed using a partial least squares-discriminant analysis (PLS-DA). PCA of the LC-MS fingerprints revealed three groups. Discriminating signals were compared to literature data and identified as two bisabolane derivatives responsible for discrimination of group A and C, and one ent-kaurenoic acid derivative, discriminating group B. A taxonomic relationship between a previously unidentified species and L. franchetii and Leontopodium sinense could be determined by comparing NMR fingerprints. This finding supports recent molecular data. Furthermore, Leontopodium dedekensii and L. sinense, two closely related species in terms of morphology and DNA-fingerprints, could be distinguished clearly using ¹H NMR and LC-MS metabolic fingerprinting.
authors.affiliation: Institute of Pharmacy/Pharmacognosy, Faculty of Chemistry and Pharmacy, Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, A-6020 Innsbruck, Austria. Stefan.Safer@uibk.ac.at
authors.forename: Stefan
authors.initials: S
authors.lastname: Safer
authors.forename: Serhat S
authors.initials: SS
authors.lastname: Cicek
authors.forename: Valerio
authors.initials: V
authors.lastname: Pieri
authors.forename: Stefan
authors.initials: S
authors.lastname: Schwaiger
authors.forename: Peter
authors.initials: P
authors.lastname: Schneider
authors.forename: Volker
authors.initials: V
authors.lastname: Wissemann
authors.forename: Hermann
authors.initials: H
authors.lastname: Stuppner
mesh_terms: D019659:Asteraceae / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004224:Diterpenes / Q000032:analysis* / Q000737:chemistry; D004777:Environment; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods; D055442:Metabolome; D015999:Multivariate Analysis; D018517:Plant Roots / Q000737:chemistry; D025341:Principal Component Analysis; D012717:Sesquiterpenes / Q000032:analysis / Q000737:chemistry; D013045:Species Specificity
chemical_list: D004224:Diterpenes; D012717:Sesquiterpenes