Bioanalytical method development and validation of atrasentan in human plasma using verapamil as internal standard by liquid chromatography coupled with tandem mass spectrometry
Konatham Teja Kumar Reddy, M. Akiful Haque
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.