Development of the UPLC and LC-MS/MS methods for the simultaneous determination of trifluridine and tipiracil hydrochloride from pharmaceutical preparations and characterization of degradation products by LCMS-IT-TOF

Cancer is the disease that causes the most deaths after cardiovascular diseases in the world. And its incidence in humans is increasing day by day. Colon cancer is a type of gastrointestinal cancer that ranks very high in cancer incidence and mortality. The drug named Lonsurf, which was approved by the FDA in 2015 for the treatment of colon cancer, contains the combination of Trifluridine and Tipiracil HCl active ingredients. In this study, UPLC and LC-MS/MS methods were developed for the analysis of pharmaceutical formulations containing TRI/TIP combination. The UPLC method was optimized in the presence of degradation products and a new degradation products of both active substances were detected under acidic, basic and oxidative harsh degradation conditions. Qualitative analysis of the degradation product was performed with the LCMS-IT-TOF instrument. For the chromatographic separation of both developed methods, different stationary phases including core shell and monolithic silica structure were tested. The most successful separation was achieved on the Kinetex® 2.6µm PS C18 Core-Shell (100×4.6 mm, ID 2.7 µm) column. The mobile phase is the methanol buffer mixture, and optimum separation was achieved with 10 mM ammonium acetate (pH 5.8) as a buffer. The flow rate was determined as 1 mL/min for UPLC and 0.5 mL/min for LC-MS/MS. Validation studies of the method obtained with optimum conditions and on the pharmaceutical preparation containing TRI/TIP were performed.