A Validated High Performance Thin Layer Chromatographic Method for Simultaneous Estimation of Berberine Chloride and Guggulsterone Z in Herbal Formulation

Abstract

A simple, precise, and robust high-performance thin layer chromatography (HPTLC) method was developed and validated for the determination of berberine chloride and guggulsterone Z in herbal formulation. Chromatographic separation was achieved on aluminium plates precoated with silica gel G60F254 as the stationary phase and toluene-acetonitrile-formic acid (5:3:0.5 v/v/v) as the mobile phase. Densitometric evaluation was carried out at 264 nm. The present method was validated according to ICH guidelines. The Rf value of berberine chloride and guggulsterone Z was found to be 0.40 ± 0.02 and 0.68 ± 0.02, respectively. The response in terms of peak area was found to be linear over the concentration range of 100-500 ng/spot for berberine chloride and 200-1000 ng/spot for guggulsterone Z with regression coefficient value greater than 0.995 for both the phytoconstituents. The method was validated by determining its accuracy, precision, robustness, specificity and system suitability. The method was found to be accurate, precise and robust to carry out the simultaneous estimation of berberine chloride and guggulsterone Z. The developed method was successfully applied for the simultaneous estimation of berberine chloride and guggulsterone Z in herbal formulation.