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Vol. 25 No. 8 (2013): Vol 25 Issue 8

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HPTLC Quantification and Antimicrobial Evaluation of Oleanolic Acid Isolated from Diospyros melanoxylon

https://doi.org/10.14233/ajchem.2013.14428
Kedar K. Rout
Department of Chemistry, North Orissa University, Sriramchandra Vihar, Baripada, Mayurbhanja-757 003, India
Ganeswar Sahoo
Department of Chemistry, Gandhi Institute of Engineering & Technology, Gunpur-765 022, India
Deeptanjali Sahoo
Medicinal Chemistry Department, Central Institute of Medicinal and Aromatic Plants, Lucknow-226 015, India
Rajesh K. Singh
Department of Chemistry, North Orissa University, Sriramchandra Vihar, Baripada, Mayurbhanja-757 003, India

Corresponding Author(s) : Kedar K. Rout

kd_rout@yahoo.co.in

Asian Journal of Chemistry, Vol. 25 No. 8 (2013): Vol 25 Issue 8

Abstract

The bioactive molecule, oleanolic acid was isolated from Diospyros melanoxyon leaves by column chromatography and characterized with the help of physical and spectroscopic data. An HPTLC method has been developed and validated for its quantification in D. melanoxyon leaves. Chromatographic separation of the compound was achieved on high performance TLC plates by using a binary mobile phase consisting of n-hexane:ethyl acetate (7.5: 2.5, v/v). It was quantified at its wavelength of maximum absorbance of 540 nm after derivatization with methanol-sulphuric acid reagent. The limit of detection and quantification were found to be 15 and 40 ng per spot, respectively. The linear regression analysis data for the calibration plot showed a good linear relationship between peak area and concentration in the range of 40-360 ng/spot with r2 = 0.9997. The instrumental precision was 1.18 % (CV) and the repeatability of the method was 1.87 % (CV). The method was validated for precision, recovery and repeatability as per the International Conference on Harmonization guidelines. The developed HPTLC method is accurate, precise and has been successfully applied for the assay of this bioactive molecule in D. melanoxylon leaves. Antimicrobial screenings of oleanolic acid revealed its potent activity against a gram positive bacteria viz., S. aureus and four fungal strains viz., A. niger, C. tropicalis, C. albicans and C. krusei.

Keywords

High performance thin layer chromatography Densitometry Diospyros melanoxylon Oleanolic acid Antimicrobial activity
K. Rout, K., Sahoo, G., Sahoo, D., & K. Singh, R. (2013). HPTLC Quantification and Antimicrobial Evaluation of Oleanolic Acid Isolated from Diospyros melanoxylon. Asian Journal of Chemistry, 25(8), 4649–4653. https://doi.org/10.14233/ajchem.2013.14428
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