HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH ULTRAVIOLET DETECTION AND PROTEIN PRECIPITATION AS A WAY OF QUANTITATIVE DETERMINATION OF NILOTINIB WITH AND WITHOUT INTERNAL STANDARD

Abstract

A simple, fast, and effective HPLC-DAD method for the quantification of nilotinib in blood serum with and without dasatinib as the internal standard was developed and presented in this work. A simple protein precipitation extraction method leads to satisfactory recoveries, accuracies, and coefficients of variation as 83.9–96.8%, 95.8–105.1%, and 2.1–6.1%, respectively. A Marcherey-Nagel Lichrospher 100-5 RP8 250 × 4 mm column held at 40°C, a mobile phase of 0.05 M H₃PO₄/KH₂PO₄-acetonitrile (7:3, v/v) at a flow rate of 1 mL/min, and a diode array detector operated at a wavelength of 265 nm were used for the analysis of 50 µL prepared sample injected into the HPLC. A single run was completed in 20 min. The presented method has a limit of quantification of 0.20 µM and is linear between 0.20 and 10 µM. The possibility of its use for the nilotinib analysis without an internal standard is also discussed.

Keywords:

• AMN107
• dasatinib
• HPLC-DAD
• nilotinib
• tasigna
• TKI