Reversed‐Phase High‐Performance Liquid Chromatography Method and Its Coupling to Mass Spectrometry for Deferasirox Stability Study and Degradation Product Identification

ABSTRACT Deferasirox (DFR) is a novel oral iron chelator used in the treatment of beta‐thalassemia, myelodysplastic syndromes, and other chronic anemias. Understanding the stability behavior of deferasirox in various degradation condition is crucial and has been studied under hydrolysis, oxidative, photolysis, and thermal conditions. A Phenomenex C 18 column (250 × 4.6 mm, 5 µm) was employed to achieve drug separation, with mass spectrometry (MS)‐compatible mobile phase comprising of acetonitrile and 10 mM ammonium acetate buffer (60:40, v/v). The flow rate and detection wavelength were 1.0 mL/min and 250 nm, respectively. The fully validated reversed‐phase high‐performance liquid chromatography (RP‐HPLC) stability method developed as per International Council for Harmonisation (ICH) Q2 R1 guidelines, demonstrated excellent performance metrics, including limit of detection (LOD; 30.01 ng/mL), limit of quantification (LOQ; 90.96 ng/mL), recovery (98%–100.1%), precision repeatability (1.8%), and intermediate precision (0.8%). Significant degradation was observed under alkaline, acidic, oxidative, and photolytic while the drug remained stable under thermal condition. The use of liquid chromatography–mass spectrometry (LC–MS) compatible solvents in the mobile phase enables further characterization of deferasirox and its degradation products. Addition of an alkyl radical being postulated as possible degradation mechanism under hydrolysis condition. The fully validated, stability‐indicating, mass‐compatible RP‐HPLC method can be effectively utilized commercially for quality control, and stability studies of deferasirox.