A QUALITY BY DESIGN APPROACH FOR THE DEVELOPMENT AND VALIDATION OF A REVERSE-PHASE ULTRA-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF ACEBROPHYLLINE AND N–ACETYLCYSTEINE IN BULK AND PHARMACEUTICAL DOSAGE FORM

Abstract

Objectives: The aim of this study was to develop and validate an accurate, fast, and reliable stability-indicating reverse-phase ultra-performance liquid chromatography (RP–UPLC) technique based on quality by design (QbD) for acebrophylline (ACB) and N–acetylcysteine (NAC) simultaneously in pure and tablet formulation. Methods: A QbD approach was adopted to choose the critical parameters. The method involved separation of analytes using BEH C18 column (50×2.1 mm, 1.7 μm) with mobile phase containing 0.1% (v/v) orthophosphoric acid and acetonitrile in 70:30 ratio with a flow rate of 0.3 mL/min. Detection was done at 273 nm with 1 μL injection volume and run time of 2 min. Linearity, accuracy, precision, specificity, robustness, limit of detection, and limit of quantification were validated. Results: Retention times of 0.763 and 1.014 min were obtained for ACB and NAC, respectively. The linearity was found to be excellent within a range of 2.5–15 μg/mL for ACB and 15–90 μg/mL for NAC with R2 values of 0.999 for both drugs. Recoveries of 99.86% for ACB and 99.81% for NAC were achieved with percent relative standard deviation <2%. No interference from excipients and good robustness were seen. Analytical GREEnness Metric (AGREE) software indicated moderate greenness with an AGREE score of 0.67. Conclusion: The proposed RP–UPLC method was fast, precise, accurate, economical, and can be used to determine simultaneous estimation of ACB and NAC in bulk and tablet formulation for routine quality control applications.