Bioanalytical method development and validation of atrasentan in human plasma using verapamil as internal standard by liquid chromatography coupled with tandem mass spectrometry

Abstract

A satisfactory LC-MS/MS separation and good peak symmetry for Atrasentan were obtained with Agilent, Zorbax, XDB C18 (2.1 x 50  mm ID, 5 μm), and a mobile phase containing a mixture of 5  mM Ammonium Formate buffer with 0.1% formic acid were mixed with HPLC grade Acetonitrile in the proportion of 70:30, v/v was delivered at a flow rate of 0.150 mL/min by positive ion mode (API 4000) with an injection volume of 10 µL and a run time of 3 min. Detection is performed by atmospheric pressure electrospray ionization (ESI) mass spectrometry in positive ion mode. The precursor to product ion transitions is m/z 5.11.600 to 354.04 for Atrasentan, and m/z 455.40 to 165.00 for Verapamil (Internal standard) were used for quantization. The retention time of Atrasentan and Verapamil (Internal standard) was found to be 1.68 & 0.96 min.