TLC Densitometric Method for the Development and Validation of Gallic Acid, Catechin and Chlorogenic Acid as Markers in Evicapoly-Herbal Formulation.
Rapid and simple high-performance TLC methods were developed for the quantitative estimation of Gallic acid, Catechin and Chlorogenic acid as active principals or marker constituents of poly herbal formulated Evica capsule. Identification and quantification were performed on 20 cm × 10 cm, layer thickness 0.2 mm, aluminum- backed silica gel 60 F254 HPTLC plates previously washed with methanol. Toluene-ethyl acetate-formic acid (5: 4: 1 v/v) used as a mobile phase for Gallic acid, toluene-ethyl acetate-formic acid- methanol (6: 3: 1.6: 0.4 v/v) used as a mobile phase for Catechin, and ethyl acetate-formic acid- acetic acid- water (10: 1.1: 1.1: 2.6 v/v) for Chlorogenic acid. The spots were scanned at λ = 273, 254 and 366 nm for Gallic acid, Catechin and Chlorogenic acid respectively. The suitability of this HPTLC method for simultaneous estimation of the marker constituents were proved by validation in accordance with ICH Guidelines. Determination of methods accuracy by the standard addition method at three concentration levels returned a mean recovery of 98.01 ± 0.16, 98.7 ± 0.24 and 97.5 ± 0.3 for gallic acid, catechin, and chlorogenic acid. The developed method has the advantage of being rapid and easy. Hence it can be applied for routine quality control analysis of these molecules in a poly-herbal formulation.