Determination of the Related Substances of Cetilistat by HPLC

OBJECTIVE To establish an HPLC method for determining the contents of the related substances of cetilistat. METHODS The condition of detection were as follows:Agilent Phenyl column (250 mm×4.6 mm, 5 μm), the gradient mobile phase consisted of 5 mmol·L^-1 potassium dihydrogen phosphate (adjust with 10% phosphoric acid to pH 4.0) and acetonitrile with a flow rate of 1.0 mL·min^-1, the UV detection wavelength was 226 nm. RESULTS The resolution of cetilistat and relate substances was good. The calibration curves were linear in the range of 0.049 5-1.981 0 μg·mL^-1 for impurity A (r=0.999 9); the calibration curves were linear in the range of 0.059 9-0.399 1 μg·mL^-1 for impurity B (r=0.999 9); the calibration curves were linear in the range of 0.059 6-0.397 5 μg·mL^-1 for impurity C (r=0.999 9). The average recoveries of impurity A, impurity B and impurity C were 104.2%, 107.0% and 107.8% respectively; the RSDs were 2.09%, 1.77% and 2.18%, respectively. CONCLUSION The method is simple, accurate, reliable and can be used for the quality control of the related substances of cetilistat.