Production of sarsasapogenin from tissue culture of Asparagus racemosus and its quantification by HPTLC

Abstract

To develop an alternative method for production of sarsasapogenin by plant tissue culture technique and its quantification in Asparagus racemosus and its in vitro cultures using HPTLC. Murashige and Skoog’s (MS) basal medium supplemented with various growth regulators was used for development of shoot and root calli while, hormone free MS medium was used for development of Agrobacterium tumefaciens (MTCC-532) mediated transformed shoot culture. The quantification of sarsasapogenin was done using CAMAG HPTLC system, solvent system: Chloroform: Acetone (80:20) and spraying reagent: Anisaldehyde sulfuric acid at 365 nm wavelength. The calli from shoot and root and morphogenetic shoot and root culture as well as Agrobacterium tumefaciens mediated transformed shoot culture of Asparagus racemosus was developed on MS medium. The sarsasapogenin was found present in natural plant as well as in in vitro cultures. It was estimated quantitatively by CAMAG HPTLC system which, showed that highest amount (0.133%) was found to be present in shoot tumor followed by root callus (0.127%) that are 2.59 and 2.5 times higher respectively than natural root. It can be concluded that the in vitro cultures produced by plant tissue culture technique have great potential for the production of higher amount of sarsasapogenin, which can be used for industrial scale up processes.