Summary
Plant secondary metabolites have enormous potential for research and new drug development. Many secondary metabolites have a complex and unique structure and their production is often enhanced by biotic and abiotic stress conditions. Gymnemic acid (C43H68O14), a pentacyclic triterpenoid isolated from the leaves of Gymnema sylvestre, exhibits potent inhibitory effect on diabetes. The gymnemic acid content is determined by chromatographic methods: Camag HPTLC system equipped with a sample applicator Linomat IV and TLC scanner and integration software CAT 4.0. In HPLC C18 (ODS) reverse phase column; water 486 UV detector; mobile phase, water/methanol (35:65, HPLC grade) + 0.1% acetic acid are used. Sample (20 μL) is applied with a flow rate of 1 mL/min and read at 230 nm with UV detector. The production of gymnemic acid is significantly higher in callus treated with 2,4-dichloro phenoxy acetic acid (2,4-D) and kinetin (KN). The blue light increases gymnemic acid accumulation upto 4.4-fold as compared with fluorescent light treatment and out of which 2.8 is found in leaves. Gymnemic acid is isolated from callus, grown under stress conditions followed by preparative TLC, simple and reproducible character based on HPTLC and high performance liquid chromatography.
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Bibliography
We are thankful to Prof. Shigenobu Arihara and Kazuko Yoshikawa, Kyoto Pharmaceutical University, Kyoto, Japan for providing gymnemic acid standard; Mr. S. Govindu, Technician, Central Electrochemical Research Institute, Karaikudi, Tamil Nadu, for carrying out the HPLC analysis; and Mr. Manimaran, Lecturer, JSS college of Pharmacy, Ooty, for the help rendered during HPTLC analysis.
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Ali Ahmed, A., Rao, A., Rao, M. (2009). In Vitro Production of Gymnemic Acid from Gymnema sylvestre (Retz) R. Br. Ex Roemer and Schultes Through Callus Culture Under Abiotic Stress Conditions. In: Jain, S.M., Saxena, P.K. (eds) Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants. Methods in Molecular Biology, vol 547. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-287-2_8
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DOI: https://doi.org/10.1007/978-1-60327-287-2_8
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