Development and Validation of the Ultra-Performance Liquid Chromatography Method for Quantification of Belzutifan in Pharmaceutical Dosage Form

The main objective of the present study was to develop and validate a simple, precise, sensitive, and accurate ultra-performance liquid chromatography (UPLC) method for the estimation of belzutifan in pure and dosage forms. The UPLC technique was developed by using an Acquity UPLC BEH Shield C8 (100 x 2.1 mm, 1.7 mu m) column. The developed technique was validated in compliance with the guidelines of the International Conference on Harmonization (ICH). Belzutifan was separated chromatographically with good resolutions using the mobile phase acetonitrile: buffer (40:60 v/v) at a flow rate of 0.5 mL/min, injection volume of 5 mu L, and a wavelength of 228 nm. The validated technique was found to be linear within the range 2.5-15 mu g/mL. Belzutifan detection and quantification limits were determined to be 0.3 and 1 mu g/mL respectively. The % RSD was not more than 2%, demonstrating the precision of the developed technique. Furthermore, the rate of recovery was close to 100%, confirming the method's accuracy. Minor changes in the chromatographic conditions revealed the method's robustness. The analytical method developed was precise, simple, reproducible, and sensitive. Hence, it can be used to estimate belzutifan.