Comparative phytochemical study on rhizome and tissue culture of Ruscus aculeatus L

Ruscus aculeatus L. (Ruscaceae), butcher's broom, is a known medicinal plant used in the treatment of venous diseases such as varicose veins and hemorrhoids [1]. It contains steroidal saponins having as aglycons ruscogenin and neoruscogenin [1,2]. Our study was focused on quantitative determination of steroidal sapogenins resulted by acid hydrolysis [3] of 7 samples of R. aculeatus from wild flora of South-Western Romania (Dealurile Lipovei). These were: rhizomes with roots, rhizome, roots, alcoholic extract (1:1) from rhizome with roots and shoots, callus and callus with roots obtained by in vitro tissue culture. The quantitative analysis has been carried out by HPLC-MS/MS, using ruscogenin and neoruscogenin as external standards. Chromatographic separation was performed on reverse phase SB-C18 column (Agilent). The mass spectrometer was operated using an ESI source in the positive mode and was set for isolation and fragmentation of sodium-dehydrated adduct ions of ruscogenin and neoruscogenin respectively, with m/z=431.3 and 429.3. The HPLC-MS/MS assay showed that the highest sapogenin concentration is present in the rhizome samples (0.11% ruscogenin and 0.17% neoruscogenin) and among the in vitro tissue culture samples the highest sapogenin concentration is present in shoots (0.017% ruscogenin and 0.075% neoruscogenin). On the other hand, all samples contain more neoruscogenin than ruscogenin.

Acknowledgements: This work was supported by the grant ET 3263/2005 CNCSIS.

References: [1] Bruneton J. (1993), Pharmacognosie, Phytochimie Plantes Medicinales, 2ed, Ed. Tec&Doc, Paris: 556–558. [2] Hostettman J., Marston A., (1995), Saponins, Ed. Cambridge University Press, Great Britain: 84, 298, 302. [3] Drapeau D., et. al. (1986), Planta Med. 6: 474–478.