HPTLC Based Analysis of Tylophorine from Cultures and In Vitro Regenerated Plants of Tylophora Indica- an Endangered Medicinal Plant

Author(s)

Kaur, H., Anand, M., Goyal, D.

Author's Affiliation

Abstract

Tylophora indica (Burm. f.) Merrill member of family Asclepiadaceae contains various therapeutically important alkaloids such as tylophorine, tylophorinine and tylophorinidine. These alkaloids are being used in various traditional health systems to cure number of ailments. Hence, it is worthy to develop an extraction technique to extract these important alkaloids. An efficient protocol has been established for extracting major secondary metabolite- tylophorine from in vitro regenerated plants, callus and suspension cultures of Tylophora indica. Leaf explants taken from healthy field grown plants were cultured on Murashige and Skoog’s medium supplemented with 29.4 μM naphthalene acetic acid and 4.65 μM kinetin for induction of callus. The compact green callus showed differentiation of shoots when transferred onto 8.8 µM benzyladenine. Microshoots after rooting were hardened, acclimatized and successfully established in field conditions with 90% survival rate. For tylophorine estimation, leaves of regenerated plants, callus and suspension cultures were extracted using cold extraction with acetic acid in methanol followed by acid extraction with ethyl acetate: HCl. Preliminary estimation of tylophorine from the crude extract was done using thin layer chromatography, column chromatography and fine purification was done using high performance thin layer chromatography (HPTLC). Quantitative analysis showed maximum tylophorine (80μg/ml) in the leaves of in vitro regenerated plants, followed by suspension (28.30µg/ml) and callus cultures (24.46µg/ml) respectively. The present study employing in vitro culture technology is an alternative towards the production of valuable metabolites to meet the competiveness and ever increasing demand for plant based raw material.

Keywords

Tylophora indica, tylophorine, high performance thin layer chromatography, in vitro

Cite This Article

Kaur, H., Anand, M., & Goyal, D. (2014). HPTLC Based Analysis of Tylophorine from Cultures and In Vitro Regenerated Plants of Tylophora Indica- an Endangered Medicinal Plant. International Journal for Pharmaceutical Research Scholars (IJPRS), 3(2), 91-95.