Development and Validation of RP-HPLC Method for Quantification of Bamifylline in Pharmaceutical Formulations using Analytical Quality by Design (AQbD) Principles

The current study presents a systematic approach based on Analytical Quality by Design (AQbD) principles. This strategy was employed to develop and validate a reliable reverse-phase high-performance liquid chromatography (RP-HPLC) method for the quantification of Bamifylline Hydrochloride (BMFH) in pharmaceutical formulations. A factorial design was employed to optimize the critical method parameters, including the composition of the mobile phase, flow rate, and detection wavelength, to enhance chromatographic performance. Chromatographic separation was achieved using a C۸ column (۱۰۰ × ۴.۶ mm, ۵ µm particle size) with a mobile phase of methanol and water (۸۰:۲۰ v/v) at a flow rate of ۰.۸ mL/min, and detection was performed at ۲۷۳ nm. The method was validated according to ICH Q۲(R۱) guidelines and demonstrated satisfactory linearity, accuracy, precision, and robustness, demonstrating excellent linearity (۵-۶۰ µg/mL, R² = ۰.۹۹۹۸), high accuracy (۹۹.۲۰-۱۰۰.۲۰%), and precision (RSD <۲%). Forced degradation studies have confirmed the chemical stability of BMFH under various stress conditions, with minimal degradation observed in typical storage environments. The greenness assessment of the method was conducted using the Green Analytical Procedure Index (GAPI), analytical eco-scale, and HPLC-EAT metrics, confirming the method’s favourable environmental profile with minimal energy consumption, low hazardous waste generation, and moderate solvent use. An AGREE score of ۰.۷۲, which is considered a high level of greenness, further demonstrates strong compliance with green analytical chemistry principles. Thus, the developed RP-HPLC method not only improves upon existing methods by offering enhanced analytical robustness through AQbD optimization, but also demonstrates superior environmental sustainability, as evidenced by its high AGREE score (۰.۷۲), excellent eco-scale rating (۸۳), and low solvent-related EAT burden. Future improvements targeting solvent replacement and miniaturization could further enhance the environmental credentials.

کلیدواژه ها:

Bamifylline hydrochloride ، RP-HPLC ، Factorial design ، Wavelength ، Quality Control ، Sustainability assessment metric