Development and Validation of an RP‐HPLC Method for the Rapid and Sensitive Quantification of Everolimus in Pharmaceutical and Medical Device Applications

ABSTRACT

Herein, a rapid, sensitive, and reversed‐phase high‐performance liquid chromatography (RP‐HPLC) method was developed and validated for the quantification of everolimus. The method utilizes a reverse‐phase Zorbex SB C18 column with a mobile phase consisting of acetonitrile and methanol, that requires a total run time of ≤ 30 min. Everolimus was detected at a wavelength of 276 nm that ensures clear separation from its related impurities, which included everolimus A, everolimus B, everolimus C, sirolimus B, and sirolimus C. Method validation was performed in accordance with the International Council for Harmonisation, ICH Q2(R1), validation of analytical procedures guidelines; which assesses critical parameters such as linearity, accuracy, precision, and robustness. The method demonstrated excellent linearity ( R ² ≥ 0.999) over a concentration range of 1–400 ppm, with recovery rates between 99%–100%. The limit of detection and limit of quantification were established at 0.01 ppm (10 ppb) and 0.1 ppm (100 ppb), respectively. The method has been optimized with the following parameters: UV wavelength at 276 nm, column temperature maintained at 55°C, run time of 30 min, ternary solvent system of methanol–water–acetonitrile maintained at a solvent ratio of 61.5%:26%:12.5%. This validated HPLC method provides a rapid, accurate, and reliable analytical tool for the routine quality control of everolimus in pharmaceutical and medical device applications.