Simultaneous estimation of Gallic acid and rutin in Kaishore guggulu and Vatari guggulu by HPTLC

Abstract

A simple, rapid, robust and reliable HPTLC method has been developed for simultaneous estimation of gallic acid and rutin in Kaishore Guggulu and Vatari Guggulu. Identification and quantification were performed on 20cm x 10cm, layer thickness 0.2mm, aluminium-backed silica gel 60 F254 HPTLC plates previously washed with methanol. Toluene: Acetone: Ethyl Acetate: Formic Acid: Water (2: 3: 2: 1: 2, v/v), was used as a mobile phase. Calibration plots were established showing the dependence of response on the amount chromatographed. The validated calibration range was 400-800ng per spot (R2 = 0.998; 0.996). The spots were scanned at λ = 312 nm. The suitability of this HPTLC method for simultaneous estimation of the marker constituents was proved by validation in accordance with ICH Guidelines. Determination of method accuracy by the standard addition method at three concentration levels returned a mean recovery of 98.92 ± 0.16–101.61 ± 0.24. The developed method has the advantage of being rapid, robust and easy. Hence, it can be applied for routine quality control analysis of gallic acid and rutin in those Ayurvedic formulations.