Comprehensive Metabolome Analysis of Fermented Aqueous Extracts of Viscum album L. by Liquid Chromatography−High Resolution Tandem Mass Spectrometry

Abstract

Fermented aqueous extracts of Viscum album L. are widely used for cancer treatment in complementary medicine. The high molecular weight compounds viscotoxins and lectins are considered to be the main active substances in the extracts. However, a vast number of small molecules (≤1500 Da) is also expected to be present, and few studies have investigated their identities. In this study, a comprehensive metabolome analysis of samples of fermented aqueous extracts of V. album from two host tree species (Malus domestica and Pinus sylvestris), both prepared by two pharmaceutical manufacturing processes, was performed by liquid chromatography−high resolution tandem mass spectrometry (LC-HRMS/MS). A total of 212 metabolites were putatively annotated, including primary metabolites (e.g., amino acids, organic acids, etc.) and secondary metabolites (mostly phenolic compounds). A clear separation between V. album samples according to the host tree species, but not due to manufacturing processes, was observed by principal component analysis. The biomarkers responsible for this discrimination were assessed by partial least squares−discriminant analysis. Because V. album extracts from different host trees have different clinical applications, the present work highlights the possibility of characterizing the metabolome for identification and traceability of V. album fermented aqueous extracts.

Untargeted metabolomics approach to discriminate mistletoe commercial products

“…Annotations were based on interrogation of public databases (GNPS 25 , MS-Dial 26 and ISDB 27 ) from MetGem software leading to annotation at level 2 28 . The annotation is in good agreement with the previous report of metabolome of fermented aqueous extracts of Viscum album L. 29 , including a large amount of small peptides (second largest cluster in Fig. 2 ) together with data compiled with Urech and Baumgartner 7 where various families of phenolic compounds were reported including derivatives of cinnamic, sinapinic, rosmarinic acids or flavones.…”

“…On the contrary, glycosylated flavanones were more abundant in Abnoba and Helixor products but not detected in Iscador/Weleda ones. Therefore, molecular networks based on MS (relative quantification) and MS/MS (annotation) data allow distinguishing the composition of Viscum album L. 29 from the three producers confirming the trends obtained by PCA analysis on MS features.…”

Metabolomics by UHPLC-Q-TOF Reveals Host Tree-Dependent Phytochemical Variation in Viscum album L.

“…album samples ( Table 2 ). These compounds were also previously described in the Viscum album species [ 18 , 20 , 21 , 22 , 23 ]; however, they were not related to the differentiation of the three clusters that were described in the PLS-DA analysis.…”

“…The metabolome analysis is mainly based on nuclear magnetic resonance (NMR) and liquid chromatography coupled to mass spectrometry (LC-MS) methodologies. A vast number of small molecules, derived from primary and secondary Viscum album metabolism, was previously described in mistletoe-fermented aqueous extracts, highlighting the sensitivity and selectivity of LC-MS for metabolite detection in complex matrices [ 18 ].…”

Phytochemical Characterization of Phoradendron bollanum and Viscum album subs. austriacum as Mexican Mistletoe Plants with Antimicrobial Activity

“…Hydroxycinnamic acids (HCAs) were the family of compounds most observed in both mistletoes. These compounds were reported as part of some mistletoes in previous investigations [ 22 , 23 ]. Lipid-lowering, antioxidant, antibacterial, and immunomodulatory effects are attributed to HCA, as in the case of L. cuneifolia known as “Creole mistletoe” or “Argentine mistletoe” [ 24 ].…”