Abstract

DEVELOPMENT OF STABILITY INDICATING REVERSED PHASEHPLC METHOD AND ITS VALIDATION FOR THE ANALYSIS OF BOTULINUM TOXIN IN API AND STERILE FORMULATION

Sonal Bhale*, Alpana Asnani and Dinesh Chaple

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Abstract

A rapid, simple, reliable and accurate reversed phase HPLC analytical method was developed for the evaluation of Botulinum toxin and the developed method was then validated as per ICH guidelines in sterile dosage form for stability studies. A C18 column was selected with a flow rate of 2 ml/min. The selected mobile phase consists of sodium phosphate buffer (0.05 M) at pH value of 2.8 and acetonitrile at the ratio of 30:70 respectively at 214 nm. Botulinum toxin peak was eluted at retention time of 2.1 min at 214 nm with total run time of 10min. Linearity and range was observed for concentration of 1μg/ml- 10μg/ml. The developed method was linear with of 0.99 as the correlation coefficient. The validation of method was done as per ICH guidelines for linearity, range, accuracy, precision, specificity, detection limit, quantitation limit, and forced degradation study.

Keywords: RP-HPLC, Botulinum toxin, method development, method validation, degradation study etc.