Abstract

OPTIMIZATION OF CALLUS BIOMASS YIELD AND EXTRACTION OF ACTIVE COMPOUND (PHYLLANTHIN) FROM LEAF AND CALLUS EXTRACT OF PHYLLANTHUS VIRGATUS G. FORST

M. R. Ramachandra Kumar, S. Ravi Kumar* and B. Janarthanam

Abstract

The objective of the present study was to evaluate the optimization of callus biomass yield and high-performance liquid chromatography (HPLC) analysis of phyllanthin compound in the leaf and callus culture extract of Phyllanthus virgatus. Callus cultures were established from nodal and leaf explants. Leaf explants showed better callus initiation than nodal explants. Maximum callus induction was observed in Murashige and Skoog (MS) medium containing 4.52μM 2,4-D. Further screening of callus culture was carried out on MS medium supplemented with different concentrations and combinations of 2, 4- D, NAA, IAA, (BA) and KN individually and in combinations. Optimum callus biomass of 14.90 ± 0.30 g/L dry weight (163.4 ± 0.20g/L fresh weight) was developed on MS media containing 4.52μM 2, 4-D, 4.65μM KN and 1.11μM BA. The harvested callus biomass was subjected to extraction and purification of phyllanthin compound. In this study, cell biomass extracts were compared with extracts from leaves of mother plants of Phyllanthus virgatus. HPLC analysis of these extracts showed that the main components of the active principles namely phyllanthin were present in sufficiently large amounts in the undifferentiated cultured cells.

Keywords: Phyllanthus virgatus, Callus biomass, phyllanthin, HPLC analysis.