ABSTRACT:
Belumosudil is mainly used to treat chronic graft-versus-host disease (cGVHD). A stability-indicating reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed to assess the stability of belumosudil in tablet dosage form. Chromatographic separation was performed using a Thermo Scientific HPLC system. The Thermo Scientific Hypersil C18 (250 x 4.6mm, 5µm) column was used and maintained at a temperature of 35?. Isocratic elution was performed with 25mM KH2PO4 buffer (pH 3.0) and acetonitrile in a ratio of 60:40v/v as mobile phase, at a flow rate of 1.0ml/min with 20µl injection volume. The detection was carried out at a wavelength of 250nm. The developed RP-HPLC method resulted in a suitable retention time of 5.1 minutes for belumosudil. The optimized method was validated according to ICH Q2 (R1) guidelines and demonstrated linearity for the concentration range of 10 - 150µg/ml (R² = 0.9995), with good precision (% RSD < 2.0%) and accuracy (101.03% recovery). The LOD and LOQ were found to be 2.58µg/ml and 7.82µg/ml, respectively. A forced degradation study was also conducted to identify degradation products by exposing the drug to acidic, alkaline, oxidative, photolytic and thermal degradation conditions. The data acquired showed that the degradation product was successfully separated without any interference, which confirms the stability-indicating nature of the developed method. The optimized method was suitable for analyzing the stability of belumosudil in tablet dosage form. The simple, accurate, precise, economical, reliable, and easy-to-use RP-HPLC method has been successfully developed and validated.