Mallikarjuna Rao N, Bagyalakshmi J, Ravi TK
Mallikarjuna Rao N.1*, Bagyalakshmi J.2 and Ravi T.K.2
1Research Scholar of JNTUK, Department of Pharmaceutical Sciences, Kakinada
2Department of Pharmaceutical Analysis, College of Pharmacy, Sri Ramakrishna Institute of Paramedical Sciences, New Siddapudur, Coimbatore, Tamilnadu India.
Volume - 4,
Issue - 1,
Year - 2011
A simple, fast, specific and precise High Performance Thin Layer Chromatographic method (HPTLC) has been developed for estimation of cholesterol standard and egg cholesterol. The chromatographic separation was carried out on precoated silica gel 60F254 aluminum plates using a mixture of toluene: acetone: glacial acetic acid (6:1.3:0.1v/v/v) as mobile phase and densitometric evaluation of spots was carried out at 215nm using Camag TLC scanner III with CATS 1.3.4 version software. The experimental parameters like band size of the spot application, chamber saturation time, solvent front migration, slit width etc. were critically studied and optimum conditions were evolved. The drug was satisfactorily resolved with Rf value 0.55±0.03. The accuracy and repeatability of the proposed method was ascertained by evaluating various validation parameters like linearity (5 to 25 µg/spot), precision (intra-day RSD 0.57 to 0.86%, inter-day RSD 0.58 to 0.90%), accuracy (96.01±0.20), and specificity according to ICH guidelines. The limits of detection and quantification were 1µg/spot and 3µg/spot respectively. HPTLC method provides a faster and cost effective method for the estimation of cholesterol.
Cite this article:
Mallikarjuna Rao N, Bagyalakshmi J, Ravi TK. Development and Validation of an Analytical Method for the Estimation of Standard Cholesterol and Egg Cholesterol by High Performance Thin Layer Chromatography (HPTLC). Research J. Pharm. and Tech. 4 (1): January 2011; Page 155-159.