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E-Journal of Chemistry
Volume 8, Issue 3, Pages 1444-1450

HPTLC Method Development and Validation for Determination of Rutin in Flavanoidal Fraction of Hibiscus micranthus Linn.

K. Ashok Kumar,1 S. Ramachandra Shetty,2 and Laxmi Narasu3

1Department of Pharmacognosy, Bharat Institute of Technology Pharmacy, Ibrahimpatnam, RR Dist-510501, India
2Department of Pharmacology, College of Clinical Pharmacy, King Faisal University, Alahsa-31982, India
3Department of Biotechnology, Jawaharlal Nehru Technological University, Kukattpally, Hyderabad-517132, Andhra Pradesh, India

Received 24 March 2010; Accepted 30 May 2010

Copyright © 2011 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A simple, precise and accurate high-performance thin-layer chromatographic method has been established for the determination of rutin in the stem extract of Hibiscus micranthus Linn. A flavonoid fraction of methanolic extract of the stem powder was used for the experimental work. Separation was performed on silica gel 60 F254 HPTLC plates with n-butanol: acetic acid: water: ammonia 3:1:1:1(v/v), as mobile phase. The determination was carried out using the densitometric absorbance mode at 286 nm. Rutin response was linear over the range 50–400 ng/μL. The concentration of rutin in the flavanoidal fraction was found to be 9.93%. The HPTLC method was evaluated in terms of sensitivity, accuracy, precision and reproducibility.