COMPARATIVE PHYTOCHEMICAL ANALYSIS OF IN-VIVO AND IN-VITRO DERIVED COSTUS PICTUS D. DON PLANTS BY HPTLC

Abstract

The present study has been conducted for micro propagation followed by the investigation of phytochemical constituents by preliminary phytochemical screening and chromatographic analysis through TLC (Thin Layer Chromatography) and HPTLC (High Performance Thin Layer Chromatography) from the leaf extract of in-vivo and in-vitro derived Costus pictus plants. The nodal explants with axillary buds of Costus pictus were cultured in B5, MS, SH basal nutrient media supplemented with various concentration of plant growth regulators (PGR’s) such as BAP, 2,4-D, IBA 1 mg to 10 mg/l, different combination of TRIA 1 $\mu$g/l to 10 $\mu$g/l. The best response was observed in 25 $\mu$g/l TRIA within 4 weeks of inoculation, complete plantlet was obtained by 8 weeks, multiple roots up to 20 cm long with pseudo stem and 6-7 board leaves. The main aim of this work was to elucidate the various secondary metabolites such as alkaloids, flavonoids, saponin and phenols in the Hexane, Di-chloro-methane, Chloroform, Ethyl acetate, Methane and Aqueous extracts of Costus pictus leaves. Initially by preliminary phytochemical screening, later TLC plating on specific mobile phase followed by advanced techniques like HPTLC on silica gel 60 F 254 plates as stationary phase. The linear ascending development was carried out in a twin trough glass chamber saturated with a mobile phase, HPTLC CAMAG “Linomat 5 sample applicator, TLC scanner3, Re prostar 3 and win CATS 1.3.4 software, spectro-densitometric scanning at 254 and 366 nm The HPTLC fingerprint profile displayed the presence of alkaloids, Flavonoids, Phenols and Saponins with reference to respective standards.