Qualitative determination of bioactive metabolites through Q-TOF LC/MS in different parts and undifferentiated cultures of Ulmus wallichiana Planchon

Abstract

Ulmus wallichiana (UW) is an endangered and folk medicinal plant used for healing fractures in India. Recently, the secondary metabolites isolated and identified from its bark revealed osteogenic activity. Considering the medicinal properties of its bark and the source for its identified and isolated metabolites, the requirement for production of alternative pathways was crucial. This study aimed to determine its bioactive metabolite using Q-TOF LC/MS (Quadrupole time-of-flight liquid chromatography- mass spectroscopy) in different parts of this plant with seasonal and altitudinal variability pertaining with the development of a protocol for induction of callus cultures and biomass production. Plant samples were collected during December - March, April - July and August - November in the year 2010-2012 from three different altitudinal locations (Nainital, Almora and Bageshwar districts) of Kumaon region, Uttarakhand, India. The results obtained from Q-TOF LC/MS analysis demonstrated the presence of four bioactive metabolites known for osteogenic activity in bark and stem samples round the year. Syntheses of bioactive metabolites in leaf samples were significantly affected by sampling season as well as altitudinal locations respectively. Callus derived from leaf explants was maintained in Murashige & Skoog (MS) medium containing (0.25 mgl-1) 2,4-Dichlorophenoxy-acetic acid (2,4-D) + (0.05 mgl-1 ) Kinetin (Kn) and (0.25 mgl-1) α-Naphthaleneacetic acid (NAA) + (0.5 mgl-1) 6- Benzylaminopurine (BAP) in liquid phase showed optimal conditions for accumulating biomass and synthesis of identified bioactive metabolites.