Introduction:
Belzutifan is a hypoxia-inducible factor-2 alpha (HIF-2α) inhibitor used for the treatment of von Hippel–Lindau disease-associated renal cell carcinoma.
Method:
A simple, accurate, precise, and specific HPLC method was developed for the estimation of Belzutifan (BZT) in human plasma using Emtricitabine (ETC) as an internal standard. The analyte and ISTD were separated on a Kromasil C18 (250x4.6mmx5µ) column using a mobile phase composition. The buffer was composed of Acetonitrile(60:40). The RTs of the analyte and ISTD were found to be 3.446 and 2.363min, respectively, at a flow rate of 1ml/min.
Results:
Further, the reported method was validated as per the USFDA guidelines, and was found to be well within the acceptable range for all parameters with concentrations of LLOQ 0.075µg/ml, LQC 0.225µg/mL,MQC 1.5µg/mL, HQC 2.4µg/mL, and ULOQ 3.0 µg/mL. The matrix effect at HQC and LQC was 100.19 and 99.83%; the sensitivity at LLOQ was 99.63%; the precision and accuracy at HQC, MQC, LQC, and LLOQ was between 98.56 and100.11%. The linearity concentration is in the range of 0.75-3µg/mL for Belzutifan with a correlation coefficient of r2 = 0.999 with good stability.
Conclusion:
The proposed HPLC method was simple, rapid, precise, and accurate for the determination of Belzutifan in human plasma. Sample preparation showed high recovery and this method shows a higher sample throughput due to the short chromatography time ( 3.446 min) and simple sample preparation. Thus, the developed HPLC method can be applied to the bioequivalence and pharmacokinetic studies of Belzutifan in human plasma samples and is appropriate for therapeutic drug monitoring in clinical laboratories.
