Development and validation of a simple stability-indicating HPLC method for the quantitation of berberine in pharmaceuticals, ayurvedic, homeopathic products and novel nanoformulation

Abstract

Berberine has been shown to have therapeutic advantages in the management of different diseases. Due to its actions and therapeutic applications, in compliance with “The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use”. (ICH) recommendations; a quick, precise and specific HPLC method was developed and validated. The chromatographic separation was carried out with Column C18 (4.6 mm X 250 mm), (5 microns) using 0.1% orthophosphoric acid: acetonitrile (60:40) as mobile phase, with a flow rate of 1 ml/min and at a wavelength of 349 nm. The developed method was also used for the estimation of berberine in various commercial preparations and also with formulated berberine-loaded cubosomal nano formulation according to ICH guidelines. Furthermore, forced degradation studies of berberine were investigated. The novel analytical technique was identified to be linear over the concentration ranging from 2 μg/ml to 14 μg/ml with R2 > 0.997. The method developed was accurate (with % recovery of 98.24% to 99.31%), precise (%RSD < 2.0%), and robust. The limit of quantification and limit of detection for berberine was found to be 0.064 μg/ml and 0.021 μg/ml respectively. In the presence of degradation products, the drug peak was obtained. The peaks for berberine were detected in the marketed formulations containing berberine and hence, the developed method could be applied to the estimation of berberine in berberine-containing formulations.