Open AccessValidation of the method for the simultaneous estimation of bioactive marker gallic acid and quercetin in Abutilon indicum by HPTLC
Author(s) -
Md. Sarfaraj Hussain,
Sheeba Fareed,
Mazhar Ali,
Arafatur Rahman
Publication year - 2012
Publication title -
asian pacific journal of tropical disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.208
H-Index - 33
ISSN - 2222-1808
DOI - 10.1016/s2222-1808(12)60127-3
Subject(s) - gallic acid , detection limit , chromatography , formic acid , quercetin , chemistry , silica gel , absorbance , ethyl acetate , toluene , organic chemistry , antioxidant
Objective: To establish and validate an simultaneous estimation of the two biomarker compounds\udgallic acid (GA) and quercetin (QE) from methanolic extract of Abutilon indicum (AI). Methods:\udChromatography was performed on aluminium foil-backed silica gel 60 F254 HPTLC plates with\udthe binary mobile phase toluene-ethyl acetate-formic acid (5:4:1, v/v/v). Ultraviolet detection was\udperformed densitometrically at the maximum absorbance wavelength, 270nm. The method was\udvalidated for precision, recovery, robustness, specificity, and detection and quantification limits,\udin accordance with ICH guidelines. Results: The system was found to give compact spots for GA\udand QE (Rf value of 0.31 and 0.50, respectively). The limit of detection (23 and 41 ng band-1) limit\udof quantification (69 and 123 ng band-1), recovery (99.4-99.9 and 98.7-99.4%), and precision (≤ 1.98\udand 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for GA\udand QE were 100-1000 (r2= 0.9991) and 150-900 ng band-1 (r2= 0.9956) and the contents estimated\udas 0.69% ± 0.01% and 0.57% ± 0.01% w/w respectively. Conclusions: This simple, precise and\udaccurate method gave good resolution from other constituents present in the extract. The method\udhas been successfully applied in the analysis and routine quality control of herbal material and\udformulations containing AI