Development and validation of RP-HPLC method for simultaneous quantification of the anticancer agents, nilotinib and sorafenib: Application in In-vitro analysis

In this research, a reversed phase high performance liquid chromatographic method was developed for the simultaneous determination of two tyrosine kinase inhibitors, nilotinib and sorafenib. Separation was performed on an Agilent C18 column (4.6×250 mm, 5µm) with mobile phase composition of potassium dihydrogen phosphate buffer (25 mM, pH 4.2) and acetonitrile (35:65 v/v) at 1.2 mL/min with UV detection at 265 nm. Specificity, linearity, precision, accuracy, and robustness of the proposed method were all assessed. Nilotinib and sorafenib had estimated retention times of 5.1 and 5.9 minutes, respectively. Linear concentration ranges for nilotinib and sorafenib, were determined as 0.05-1 µg/mL and 10-45 µg/mL with comparable coefficient correlations (0.999). For nilotinib and sorafenib, the limits of detection (LOD) were determined as 0.030 and 0.020 µg/mL, while the limits of quantification (LOQ) were 0.101 and 0.069 µg/mL respectively.