Characterization of an Unknown Impurity in Indobufen Tablets by HPLC-Q-TOF MS and NMR

Abstract

Background: Indobufen is a drug that hinders the aggregation of platelets by reversibly repressing the cyclooxygenase enzyme, further bringing about diminished thromboxane production. During quality control of indobufen tablets, an unknown impurity was detected. Objective: To characterize an unknown impurity in indobufen tablets. Method and Results: A new method compatible with mass spectrometry detection was set up. A C18 column at 35 °C with a mobile phase consisting of aqueous buffer (including ammonium formate) and methanol (35: 65, v/v) was used at a flow rate of 1.0 mL/min at 228 nm. High-performance liquid chromatography quadrupole time-of-flight mass spectrometry mass spectrometry (HPLC-Q-TOF MS) was used to identify the impurity with the electrospray ionization (ESI) source in the positive ionization mode. The results of HPLC-Q-TOF MS analysis indicated that the protonated molecule ions [M + H]+ of the unknown impurity was at m/z 312. Preparative LC method was put into practice with a Prep-C18 column with a mobile phase consisting of water and methanol (20: 80, v/v) at a flow rate of 20.0 mL/min at 228 nm. The assignment of the 1D and 2D NMR signals was performed for the unknown impurity. In addition, possible formation of the novel impurity was also studied. Conclusion: An unknown impurity in indobufen tablets was characterized. The impurity was assigned as 2-(4-(1- hydroxy-3-oxoisoindolin-2-yl) phenyl) butanoic acid.