Biological activity and phytochemical analysis of homeopathic preparations of Viscum album L

  • Carla Holandino Quaresma UFRJ
  • João VC Batista Multidiciplinary Laboratory of Pharmaceutical Sciences,
  • Michelle NO Melo Multidiciplinary Laboratory of Pharmaceutical Sciences
  • Fortune Homsani Multidiciplinary Laboratory of Pharmaceutical Sciences
  • Adriana P Oliveira Multidiciplinary Laboratory of Pharmaceutical Sciences
  • Leandro M Rocha Fluminense Federal University
  • Venicio Feo da Veiga Institute of Microbiology
  • Daniela S Alviano Institute of Microbiology
  • Celuta S Alviano Institute of Microbiology
  • Mirio Grazi Society for cancer Research
  • Hartmut Ramm Society for cancer Research
  • Matthias Kunz Society for cancer Research
  • Konrad Urech 4Society for cancer Research
  • Ulrike Weissenstein Society for cancer Research
  • Stephan Baumgartner Society for cancer Research

Abstract

Several in vitro and in vivo studies investigated the biological properties of mistletoe preparations [1-4]. In Europe, where phytochemical constituents of aqueous Viscum album extracts were previously described, the most widespread and common form of mistletoe is the white-berried species (Viscum album) [5]. The aim of the present study was to analyze the biological activity and phytochemical features of different Viscum album ssp mother tinctures collected in 2 different harvesting periods.

Leaves, stems and berries of Viscum album samples were subjected to ethanolic extraction (45% v/v) following the homeopathic pharmacopeia [6,7]. The mother tinctures (MT) prepared were: V. album (from 3 host trees Malus domestica, Quercus sp, Ulmus sp), V. album ssp. austriacum (host tree Pinus sylvestris) and V. album ssp. abietis (host tree Abies pectinata). All samples were collected at Höfli farm (Switzerland) in July and August and subjected to chemical analysis by means of high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) and Thin-Layer Chromatography (TLC). Tumor (Yoshida and Molt4) and non-tumor cells (Ma104) were incubated with MT (1; 0.1; 0.01%) for 4, 24 and 48 hours at 37ºC. Proliferation was indirectly measured using WST-1 (tumor cells) and MTT (non-tumor cells). All cells (1.5 x 104/100 μL) were plated in triplicate in 96-well plates. The cell proliferation inhibition rate was calculated as percent inhibition relative to control cells treated with the MT solvent (45% ethanol v/v). Apoptosis/necrosis was measured using 2 x 105 cells incubated with Annexin V-FITC and 7-AAD at room temperature in the dark, and analyzed in flow cytometer [2]. Antimicrobial activity was assessed for Candida albicans, Cryptococcus neoformans, Escherichia coli and Staphylococcus aureus following the Clinical Laboratory Standards Institute manuals [8-9] and compared with 45% ethanol v/v (MT solvent).

The chromatographic assays of MT samples showed phenolic acids and lignans as main chemical classes identified. WST-1 and MTT experiments revealed different antitumor potential of MT. In general, Yoshida cells were more sensitive than Molt4. The maximal cytotoxic effect with 100% of mortality was attained by incubation of Yoshida and Molt-4 cells with 1% MT of V. album ssp. abietis (harvested in July) for 4 hours. Preliminary results with non-tumor cells indicate lower cytotoxicity compared to tumor cells. Annexin V/7ADD showed predominance of late apoptotic/necrotic events after 4 and 24 hours of incubation with all MT. The antimicrobial assay showed that MT of Viscum album ssp. austriacum (host tree Pinus sylvestris) exhibited minimum inhibitory concentration 7.25 mg/mL for the 2 tested bacterial strains.

The results of the present study suggest that Viscum album samples prepared according to the homeopathic pharmacopeia exhibit promising biological activities in a dose-dependent manner. The effects of potentiation will be assessed and compared to non-potentiated samples in future experiments. Therefore, the present study contributes with data on the main chemical components in homeopathic preparations of Viscum album, drawing attention to the relevance of extract solvent, seasonal aspects, host tree and harvested parts of plant harvested to the quality of thee mother tinctures.

Author Biographies

Carla Holandino Quaresma, UFRJ
Dr. Carla Holandino Quaresma received her Ph.D. degree in Sciences from the Federal University of Rio de Janeiro (2000). Her primary field is pharmacy with emphasis on cancer research. Dr Holandino is a PI of Multidisciplinary Laboratory of Pharmacy Sciences, at the Federal University of Rio de Janeiro, and has supervised in the last 10 years, ten post-graduate students in the field of Pharmacy. Other research has been developed under Holandino’s supervision in the fields of: microbiology; electrotherapy; homeopathy; pharmaceutical procedures; pharmaceutical compounding
João VC Batista, Multidiciplinary Laboratory of Pharmaceutical Sciences,
School of Pharmacy, Federal University of Rio de Janeiro, Brazil
Michelle NO Melo, Multidiciplinary Laboratory of Pharmaceutical Sciences
School of Pharmacy, Federal University of Rio de Janeiro, Brazil
Fortune Homsani, Multidiciplinary Laboratory of Pharmaceutical Sciences
School of Pharmacy, Federal University of Rio de Janeiro
Adriana P Oliveira, Multidiciplinary Laboratory of Pharmaceutical Sciences
School of Pharmacy, Federal University of Rio de Janeiro
Leandro M Rocha, Fluminense Federal University
Niterói, Rio de Janeiro
Venicio Feo da Veiga, Institute of Microbiology
Federal University of Rio de Janeiro
Daniela S Alviano, Institute of Microbiology
Federal University of Rio de Janeiro
Celuta S Alviano, Institute of Microbiology
Federal University of Rio de Janeiro
Mirio Grazi, Society for cancer Research
Hiscia Institute
Hartmut Ramm, Society for cancer Research
Hiscia Institute
Matthias Kunz, Society for cancer Research
Hiscia Institute
Konrad Urech, 4Society for cancer Research
Hiscia Institute
Ulrike Weissenstein, Society for cancer Research
Hiscia Institute
Stephan Baumgartner, Society for cancer Research
Hiscia Institute
Published
2018-02-16
How to Cite
QUARESMA, Carla Holandino et al. Biological activity and phytochemical analysis of homeopathic preparations of Viscum album L. International Journal of High Dilution Research, [S.l.], v. 17, n. 1, p. 25-26, feb. 2018. ISSN 19826206. Available at: <http://highdilution.org/index.php/ijhdr/article/view/900>. Date accessed: 23 may 2018.
Section
Conference Presentation