LC-MS, GC-MS and in vitro pharmacological analysis of Clematis vitalba L. upper parts

The aerial parts and roots of Clematis vitalba L. (Ranunculaceae) are traditionally used as remedy for inflammatory diseases. In this study we analyzed fresh and dried flowers, leaves and stems as well as fresh fruits. These plant organs were extracted with three different solvents (ethanol, acetone and dichloromethane (DCM)) and tested for NF-κB inhibition and PPARbeta/delta activation in HEK293 cells via luciferase-based reporter gene assays. The ethanol and the acetone extract of the fruits showed the highest activity in both assays, followed by the DCM extracts of the fresh flowers and fruits (see table 1). Phytochemical analysis of fruit extracts using LC-DAD-MSⁿ and GC-MS analysis showed very similar chemical profiles of the ethanol and acetone extracts with flavonoids and triterpenes as the major constituents. In the DCM extract compounds like a phenylalanin derivative and sterols dominated the LC-MS profile. In the fresh flower DCM extract only one main peak dominated the LC-MS chromatogram. It was tentatively identified as the isovitexin derivative vitalboside.

Tab. 1: Pharmacological activities of Clematis vitalba L. active extracts

Plant parts/ pharmacological assay	Fresh fruits			Fresh flowers	Positive control
	EtOH	Acetone	DCM	DCM
NF-κB inhibiton (% inhibition at 50 µg/ml)	98.6%	98.0%	58.5%	78.4%	75.8% (Partenolide 5µM)
PPARbeta/delta activation (fold activation at 10 µg/ml)	5.41 ± 1.48	4.81 ± 1.26	2.30 ± 0.43	1.98 ± 0.34	16.70 ± 4.01 (GW0742 20 nM)

Acknowledgements: We gratefully acknowledge the funding provided by the Austrian Science Fund (FWF) within project SS107 (Drugs from Nature Targeting Inflammation).