HSPiP and quality by design aided optimized bioanalytical method development for amphotericin B quantification from plasma samples

Abstract

Amphotericin-b (AmB) is a broad-spectrum polyene macrolide antifungal and anti-Leishmaniosis. We addressed HSPiP and QbD (quality by design) oriented HPLC (high-performance liquid chromatography) method development and validation, followed by identifying critical attributes on retention time (RT) and peak area (PA). Various quality control parameters were estimated, such as the LLOQ (lower limit of quantification) and LLOD (lower limit of detection). HSPiP and experimental solubility data were the best fit in the prediction. QbD identified acetic acid (AA) pH, column temperature, and flow rate as the prime factors having an impact on PA and RT. The optimized mobile phase included AA, acetonitrile, and methanol (05:85:10) at pH 5 fluxed into the column maintained at 30 °C. The method was sensitive, as evidenced by the low values of LLOQ (1.6 ng/mL) and LLOD (10 ng/mL) and high recovery from the plasma. The method determined pharmacokinetic parameters from the rat plasma with high accuracy, precision, robustness, sensitivity, and reproducibility.

Keywords

• AmB
• HSPiP and QbD oriented optimization
• Validation parameters
• Pharmacokinetic parameters estimated from rat plasma