The ICH guidelines in practices: Stress degradation studies on botulinum toxin and validation of developed stability-indicating HPLC method

Abstract

A fast, simple, reliable and accurate RPHPLC analytical method was developed for the evaluation of botulinum toxin, and the developed method was subsequently validated according to ICH guidelines in sterile dosage form for stability studies. A C18 column with a flow rate of 2 ml/min was selected. The mobile phase chosen consisted of sodium phosphate buffer (0.05 M) at pH 2.8 and acetonitrile in a ratio of 30:70 respectively at 214 nm. Measured at an Rt of 2.1 min. 10 minutes running time. Linearity and range were observed at concentrations from 1 µg/ml to 10 µg/ml. The method developed was linear with a correlation coefficient of 0.99. Validation of the method was performed according to ICH guidelines for assay, linearity and range, precision, limit of detection, limit of quantitation, and forced degradation test.