Journal of Analytical & Bioanalytical Techniques
Open Access

Abstract

A simple, reversed-phase high performance liquid chromatography method with mass spectrometric detection (HPLC- MS/MS) was developed for determination of Pitofenone HCl in human plasma using fenpiverinium bromide as internal standard. The procedure involves a simple protein precipitation step by mixing 0.1 mL plasma, 0.05 mL of internal standard with 1.5 mL methanol. After centrifugation, 10 μl of the supernatant was injected onto a ZORBAX ECLIPSE XDB - C-18, 4.6×150 mm, 5 μm, and eluted with a mobile phase consisting in a mixture of methanol:10 mM ammonium acetate in the ratio of 70:30. Detection was in MRM mode, using an electrospray negative ionization. The ion transition monitored was: m/z Q1 MASS 368.1; Q3 MASS 112.1(M/Z) for pitofenone HCl and Q1 MASS 338.3; Q3 MASS 239.1 (M/Z) for internal standard. The retention time of pitofenone and internal standard were 3.2 and 1.7 respectively. The 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.

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