Enhanced in vitro Production of Vasicine andVasicinone from Callus Cultures of Adhatoda vasica Nees

Adhatoda vasica (L.) Nees (family Acanthaceae) is a shrub known as Malabar nut tree (Vasaka) and has been used in the indigenous system of medicine in India for over 2000 years. The major bioactive alkaloids are vasicine and vasicinone. These are responsible for uterine stimulant, antitussive and bronchodilator activity [1]. In the present investigation in vitro callus cultures of Vasaka leaf was developed for the production of vasicine and vasicinone as an alternative source. A HPTLC method for simultaneous determination of vasicine and vasicinone was developed using CAMAG HPTLC system and wincats software, which was used for quantification from leaf material and in vitro culture. The in vitro callus cultures of Vasaka leaf was successfully developed and maintained on MS basal medium supplemented 6-BA (0.5 ppm) + IBA (1.0 ppm) for 3 months. The solvent system Ethyl acetate: Methanol: Ammonia (8: 2: 0.5) was found to give compact and well resolved spots of vasicine (R_f 0.41) and vasicinone (R_f 0.72). Densitometric analysis of vasicine was carried out in the absorbance mode at 290 nm whereas vasicinone was observed at 280 nm. The vasicine content from the leaf material was 0.17%, whereas in in vitro culture it was 0.47% w/w. The content of vasicinone in leaf material was found to be 0.0013, whereas in the callus culture it was 0.046% w/w. Reference: [1] Jahangir T, Sultana S (2006) Biol Trace Elem Res. 111: 217–28.