LC-MS/MS METHOD FOR THE QUANTIFICATION OF PITOFENONE IN HUMAN PLASMA

A simple, reverse-phase high performance liquid chromatographic method with mass spectrometric detection (HPLC-MS/MS) was developed for determination of Pitofenone in human plasma using Fenpiverinium Bromide as an internal standard. The procedure involves a simple protein precipitation technique using ZORBAX ECLIPSE XDB – C-18, 4.6 × 150 mm, 5µm, column. The mobile phase used was methanol: 10 mM ammonium acetate in (70: 30). Precipitation was done using methanol and detection was done in MRM mode, using an Electro Spray positive ionization. The ion transition monitored was (m/z) Pitofenone (Q1 Mass: 368.1;Q3 Mass 112.1), Internal standard (Q1 Mass: 338.3;Q3 Mass 239.1). The retention time of Pitofenone and Internal Standard were 3.2 and 1.7 respectively. Method was evaluated in terms of linearity, accuracy, precision, recovery, sensitivity. The simple extraction procedure and short chromatographic runtime make the method suitable for therapeutic drug monitoring studies.