AQbD-Assisted Development and Validation of a Green RP-HPLC Method for Simultaneous Quantification of Telmisartan, Amlodipine Besylate, and Bisoprolol Fumarate
Year: 2026
Type: preprint
Abstract: The simultaneous determination of Telmisartan (TEL), Amlodipine besylate (AML), and Bisoprolol fumarate (BIS) presents a significant analytical challenge owing to their markedly different physicochemical properties, including variations in lipophilicity and ionization behavior. In the present study, a novel Analytical Quality by Design (AQbD)-driven reverse-phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated for the simultaneous quantification of TEL, AML, and BIS in bulk and synthetic mixtures. Critical chromatographic variables, including acetonitrile content, buffer pH, and flow rate, were systematically optimized using a Box–Behnken Design to establish a robust analytical design space and improve method understanding. The optimized chromatographic separation was achieved on a C18 column (250 × 4.6 mm, 5 µm) using phosphate buffer and acetonitrile (60:40, v/v) adjusted to pH 3.5, delivered at a flow rate of 1.0 mL min⁻¹ and column temperature of 35°C. Under optimized conditions, BIS, AML, and TEL were eluted at 3.56, 6.38, and 12.63 min, respectively, with satisfactory peak symmetry (tailing factor ≤ 1.4) and column efficiency (3754–6725 theoretical plates). Statistical analysis demonstrated excellent model adequacy, with adjusted R² values ranging from 0.8737 to 0.9921 and adequate precision values between 16.01 and 66.05, confirming the reliability of the developed AQbD model. The method exhibited excellent linearity over concentration ranges of 1–4 µg mL⁻¹ for BIS, 2–8 µg mL⁻¹ for AML, and 16–64 µg mL⁻¹ for TEL, with correlation coefficients of 0.9990, 0.9988, and 0.9988, respectively. Recovery studies demonstrated high accuracy, with recoveries ranging from 99.55–101.11% for BIS, 99.64–100.68% for AML, and 99.23–102.82% for TEL. Precision studies showed %RSD values below 2% for repeatability, intraday precision, and intermediate precision, confirming excellent method reproducibility. Deliberate variations in chromatographic parameters produced negligible effects on analytical performance, indicating robust method behavior. Furthermore, greenness assessment using the AGREE metric yielded a score of 0.66, demonstrating acceptable environmental sustainability of the developed procedure. The proposed AQbD-assisted RP-HPLC method offers a reliable, precise, accurate, and environmentally conscious analytical platform for the simultaneous determination of TEL, AML, and BIS. The integration of statistical optimization and green analytical principles provides enhanced method robustness and supports its application in routine pharmaceutical quality control and future industrial analyses.
Source: Research Square
Institutions: Parul University, Gujarat Technological University
Language: English
Subfield: Analytical Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W7165777368