Analysis of ivacaftor and determination its degradationproducts with using various liquid chromatography techniques

Cystic fibrosis is a life-threatening genetic disease that causes serious damage to the lungs. Ivacaftor, the first drug to target the underlying defect of the disease caused by specific mutations, is a concrete example of the potential of sensitive medicine. Clinical trial and registration studies have shown that Ivacaftor improves outcomes and reduces hospitalizations. In this study, a number of new analytical methods have been developed and validated for quantitative determination of Ivacaftor and qualitative determination of degradation products. The quantitative determination of Ivacaftor was performed by LC-DAD and LC-MS/MS, and qualitative determinations were performed by LCMS-IT-TOF. The validation studies of the developed methods were performed according to ICH Q2(R1) regulations. Separation of Ivacaftor and its degradation products was performed using the same chromatographic conditions; where the separation was achieved with Kinetex® (150 × 3 mm 2.6 µm, Phenomenex Co.) stationary phase functionalized with C18 group. The mobile phase conditions were 0.1% (v/v) formic acid in water and 0.1% (v/v) formic acid (27:63 (v/v) in acetonitrile, pH = 2.5; the flow rate 0.25 mL/min was used for the all methods. In the proposed method, recovery studies were performed in the solutions which are compatible with the matrix one to one. In the degradation studies, a total of five degradation products have been identified, three of which have never been reported in the literature up to date, and two of which have been synthesized for various purposes and obtained CAS number. Their structures were confirmed by high resolution mass scanning analysis using LCMS-IT-TOF. The proposed protocols are methods that can be implemented in almost any analytical laboratory. Keywords: Ivacaftor, HPLC, LC-DAD, LC-MS/MS, LC/MS-IT-TOF