Journal of Analytical & Bioanalytical Techniques
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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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