DEVELOPMENT OF A DENSITOMETRIC HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHIC METHOD FOR THE QUANTITATIVE ANALYSIS OF URSOLIC ACID IN THE LEAVES OF SPECIES OF GENUS TECOMA AND TABEBUIA OF BIGNONIACEAE FAMILY
Keywords:Bignoniaceae, Tecoma, Tabebuia, Ursolic acid, High-performance thin layer chromatography
Objective: A simple and sensitive high-performance thinâ€“layer chromatographic (HPTLC) method was developed for the quantification of ursolic acid in the leaves of three species of genus Tecoma (Tecoma gaudichaudi DC, Tecoma capensis (Thunb.) Lindl, Tecoma stans (L.) Juss. ex Kunth) and genus Tabebuia (Tabebuia rosea Bertol) belong to family Bignoniaceae.
Methods: Chromatography was performed on Silica gel 60 F254 precoated HPTLC Plates with optimised mobile phase pet ether: ethyl acetate: formic acid (7:3:0.5, v/v/v). The plate was derivatized with p-anisaldehyde reagent and scanned at 540 nm. The developed method was found to give a compact spot for ursolic acid at Rf value 0.43Â±0.01. The method was validated using International Council for Harmonization (ICH) guidelines, including linearity, precision, accuracy, and robustness.
Results: Ursolic acid was found to be present in four species, i.e., Tecoma gaudichaudi DC (1.48%w/w), Tecoma capensis (Thunb.) Lindl. (0.79%w/w), Tecoma stans (L.) Juss. Ex Kunth (1.11%w/w), Tabebuia rosea (Bertol.) (1.13%w/w). A good linearity relationship was found to be (200-1400ng bandâˆ’1) with a correlation coefficient (r2) value of 0.9946 with ursolic acid. Limit of detection and limit of quantification was considered to be 40.66, 123.21 ng per band respectively. The developed method was found to be accurate and precise with 1.32%, 1.19% (%RSD) for interday and intraday precision. The accuracy of the method was performed by recovery studies at three different concentration levels, and the average percentage recovery was found to be 98.05% for ursolic acid.
Conclusion: The proposed method for the quantitation of ursolic acid was found to be reproducible and simple.
Anonymous. The wealth of India: raw materials, New Delhi Publications and Information Directorate, C.S.I.R; 2003.
Kirtikar KR, Basu BD. Indian Medicinal plants, International Book distributors Dehradun; 1999.
Rahmatullah M, Samarrai W, Rownak J, Shahnaz RSZUM, Nasima. An ethnomedicinal, pharmacological and phytochemical review of some Bignoniaceae family plants and a description of bignoniaceae plants in folk medicinal uses in Bangladesh. Adv Natr Appl Sci 2010;4:236-53.
Connolly JD, Hill RA. Triterpenoids. Natu Prod Rep 2000;17:463â€“82.
Allouche Y, BeltrÃ¡n G, Gaforio JJ, Uceda M, Mesa MD. Antioxidant and antiatherogenic activities of pentacyclic triterpenic diols and acids. Food Chem Toxi 2010;48:2885â€“90.
Choudhary S, Datta S, Talukdar AD. Phytochemistry of the family bignoniaceae-a review. Assam Univ J Sci Technol Biol Environ Sci 2011;7:145-50.
Shetty P, Mangaonkar K, Sane RT. HPTLC determination of ursolic acid in Alstonia scholaris R. Br J Planer Chromato-Modern TLC 2007;20:65-8.
Raut K, Dash B, Singh R. HPTLC quantification and antimicrobial activity of ursolic acid from Diospyros melanoxylon. J Planer Chromato-Modern TLC 2012;25:320-5.
Dixit P, Pal M, Upreti D. Comparative studies on the analytical and antioxidant activities of the medicinally important stem bark of Holoptelea integrifolia. J Planer Chromato-Modern TLC 2014;27:162-5.
Liu J. Pharmacology of oleanolic acid and ursolic acid. J Ethnoparmacol 1995;49:57-68.
Salvador JAR, Leal ASM, Santos RC. In: Pentacyclic Triterpenes as Promising Agents in Cancer. Salvador JAR. editor. Nova Science Publishers, Inc; New York; 2010.
Novotny L, Vachalkova A, Biggs D. Ursolic acid: an anti-tumorigenic and chemopreventive activity. Neoplasma 2001;48:241-6.
Wagner H, Bladt S. Plant drug analysis. 2nd ed. Springer-Verlag Berlin and Heidelberg; 1996.
International Council for Harmonization (ICH) HARMONISED TRIPARTITE GUIDELINE. Validation of analytical procedures: Methodology; 2005. p. 1-13.
Shrikant B, kirti L. Synthesis and characterization of new imine and pthalic acid derivatives of ursolic acid. Int J Pharm Pharm Sci 2014;10:560-4.