Qualitative and quantitative analysis of Potentilla fulgens roots by NMR, matrix-assisted laser desorption/ionisation with time-of-flight MS, electrospray ionisation MS/MS and HPLC/UV.

INTRODUCTION: Potentilla fulgens is a commonly used folk medicine by natives of northeast India, Nepal and Bhutan and is rich in polyphenolic and triterpene constituents. OBJECTIVE: To identify chemomarkers in the roots of P. fulgens by an interplay of (13)C-NMR, matrix-assisted laser desorption/ionisation with time-of-flight (MALDI/TOF) MS, electrospray ionisation (ESI) MS/MS and HPLC/UV. MATERIAL AND METHODS: The (13)C-NMR spectrum of crude methanolic extract was recorded in deuterated dimethyl sulphoxide. For MALDI/TOF/MS analysis, 2,5-dihydroxybenzoic acid was used as the matrix. For determination of chemical constituents, two independent simple isocratic HPLC/UV methods for monomeric/oligomeric flavanols and triterpene acids were developed and validated. RESULTS: The (13)C-NMR spectrum of the methanolic extract indicated the presence of B-type oligomeric polyphenolics containing mainly epicatechin/catechin (epicat/cat) and epiafzelechin/afzelechin (epiafz/afz) as the monomeric units. Several isobaric monomeric and oligomeric flavanols and triterpenoids were tentatively identified by MALDI/TOF/MS and ESI/MS/MS. Fourteen compounds (four monomeric and five dimeric flavanols and five triterpene acids) were isolated using repeated column chromatography and semi-preparative HPLC, and were quantitated using HPLC/UV. CONCLUSION: It is evident from these analyses that roots of P. fulgens contain flavans, including oligomeric flavanols, as major constituents followed by triterpene acids. The methods described can be applied to other Potentilla species to identify their constituents.

Two new stereoisomeric antioxidant triterpenes from Potentilla fulgens.

The roots of Potentilla fulgens have been used for a long time as a folk remedy for many ailments without having information on its pharmacological action. Of the various extracts prepared by partitioning of the methanol extract, the ethyl acetate fraction was found to possess better antioxidant and cytotoxic activities. The degree of reduction in cloning efficiencies of MCF-7 cell lines was more with ethyl acetate than with hexane fraction of the root-extract. Hence, this fraction was further purified and nine compounds, including two new ursane type triterpenoids Fulgic acid A (4) and Fulgic acid B (5), were identified and characterized. Other compounds were identified as ursolic acid, euscaphic acid, corosolic acid, epicatechin, catechin, p-hydroxybenzaldehyde and gallic acid. Chemical structures were elucidated by spectroscopic methods, especially ESIHRMS and 2D NMR techniques. The new compounds showed good antioxidant activity and therefore this plant can be a source of natural antioxidants.

Comparison of effectiveness in antitumor activity between flavonoids and polyphenols of the methanolic extract of roots of Potentilla fulgens in breast cancer cells.

Potentilla fulgens has been used for a long time as folk remedy for a variety of ailments without having information on its pharmacological action. The study was intended to determine the effectiveness in antitumor activity among kaempferol (KMP) as flavonoids, ellagic acid (ELA) as polyphenols and methanolic extract of the root of P. fulgens (PRE) in Ehrlich ascites tumour (EA) and MCF-7 cancer cells. The total phenolic and flavonoid content were found to be 138.8 ± 1.6 mg gallic acid and 401.6 ± 4.6 mg quercetin per 1 gm of the extract, respectively. The extract resulted in increasing in vivo survivality of mice bearing EA cells and loss of cell viability in a dose-dependent manner in MCF-7 cells. This effect may be attributed to apoptotic cell death as confirmed by flow cytometric analysis and PARP1 proteolysis. Such induction of apoptosis was achieved by suppression of inhibitor of the apoptosis protein survivin. It was observed that endogenous level of glutathione was depleted significantly in MCF-7 cells after the treatment with PRE only but not with KMP or ELA. This induction of apoptotic cell death and lowering the level of glutathione are highly desirable mode for an anticancer agent.