ANALYTICAL QUALITY BY DESIGN-DRIVEN DEVELOPMENT AND VALIDATION OF A STABILITY-INDICATING RP-UPLC METHOD FOR SIMULTANEOUS DETERMINATION OF DULOXETINE HYDROCHLORIDE AND PREGABALIN IN BULK AND PHARMACEUTICAL DOSAGE FORM
Abstract
Objective: The present study aimed to validate and develop a robust, stability-indicating Reversed Phase-Ultra Performance Liquid Chromatography (RP-UPLC) method for the concurrent determination of Pregabalin and Duloxetine Hydrochloride using the Analytical Quality by Design (AQbD) Principles. Methods: An AQbD-assisted RP-UPLC method was developed using central composite design (CCD) to investigate the impact of column temperature, flow rate and mobile phase. The separation was carried out on an Acquity RP-UPLC HSS C18 column (100×2.1 mm, 1.8 µm) with an isocratic elution of 0.1% orthophosphoric acid (OPA) buffer and Acetonitrile [75:25 (v/v)] at 0.3 mL/min flow rate, and wavelength maintained at 225 nm. The developed method was performed as per International council for harmonization [ICH Q2(R1)] guidelines. Results: Chromatographic separation of Duloxetine hydrochloride and Pregabalin was achieved within a short run time, with retention times of 0.898 and 1.205 min, and satisfactory system suitability conditions. Good linearity was demonstrated across concentration ranges of 5-30 µg/mL for Duloxetine Hydrochloride and 18.7-112.5 µg/mL for Pregabalin with correlation coefficients of 0.999. Accuracy studies yielded recoveries within 98-102%, whereas precision studies showed %RSD values < 2%. Furthermore, forced degradation studies confirmed that stability-indicating ability of the developed method by ensuring effective separation of degradation drug products from the drug peaks. Conclusion: The AQbD-assisted RP-UPLC showed satisfactory accuracy, rapidity, robustness, and stability-indicating capability. Owing to its satisfactory validation characteristics and short analysis time, the developed method is effectively employed for routine quality control assessment of pharmaceutical preparations containing Pregabalin and Duloxetine Hydrochloride.