AQBD DRIVEN DEVELOPMENT AND VALIDATION OF A HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF FURAZOLIDONE AND METRONIDAZOLE IN PHARMACEUTICAL DOSAGE FORM

A Quality by Design approach to analytical method development involves selection of predetermined objective, determined critical parameters, evaluation, appropriate method selection and risk assessment. Analytical Quality by Design (AQbD) helps to understand the selected method more precisely than traditional method. An important component of AQbD is understanding of independent variables (Factors) and dependent Variables (Response) and their interactions were analysed. Present study entails AQbD driven development of reversed-phase (RP) high-performance liquid chromatography (HPLC) for simultaneous estimation of Furazolidone and Metronidazole in pharmaceutical dosage form. The risk based HPLC method development and validation for combination of pharmaceutical dosage form helps to optimize the separation process of the APIs. The chromatographic conditions were optimized using Stat-Ease 360 software ,with 32 factorial design applied to analyse the effect of mobile phase composition and flow rate and effect was evaluated on retention time, number of theoretical plates (NTP), symmetry factor and resolution. The analysis was performed on HiQSil C18H5 column (4.6mm ×250mm) using acetonitrile: phosphate buffer (pH adjusted to 3 with 0.1% OPA) (55:45 v/v) as mobile phase with flow rate 0.8 ml/min. The method achieved retention time 4.7 min for Furazolidone and 3.5 min for Metronidazole respectively. The developed method was found to be linear in the concentration range of 10-50 µg/ml with regression coefficient (r2) 0.9988 for Furazolidone and concentration range 10-50 µg/ml with regression coefficient (r2) 0.9971 for Metronidazole detection wavelength at 324 nm. The % RSD for intraday and inter-day precision was 1.34% and 1.7% for Furazolidone, and 1.34% and 1.38% for Metronidazole respectively. The robustness value was less than 2% for both APIs. The assay results were 98.6% for Furazolidone and 98.65% for metronidazole. All method validation parameters fell within the prescribed limits outlined by ICH Q2(R1). Consequently, a QbD approach was employed in the development of the RP-HPLC analytical method to enhance understanding of the interactions between independent and dependent variables.