Sensitive and specific liquid chromatography-tandem mass spectrometry method for assay of fluoxetine and its metabolite norfluoxetine in human plasma and application of method to pharmacokinetic analysis |
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Authors: | Hailin Zhu Jinwen Luo Guogang Zheng Jianzhong ShenTu |
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Institution: | (1) Department of Physiology, University Medical School of Debrecen, PO Box 22, 4012 Debrecen, Hungary;(2) Department of Pharmacology and Pharmacotherapy, Semmelweis University, PO Box 370, 1445 Budapest, Hungary; |
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Abstract: | A simple, specific and sensitive high-performance liquid chromatography — electrospray tandem mass spectrometry method is
developed for the simultaneous determination of fluoxetine and its metabolite norfluoxetine in human plasma. Plasma samples
were simply treated with acetonitrile to precipitate and remove proteins and the isolated supernatants were directly injected
into the high-performance liquid chromatography — electrospray tandem mass spectrometry system. Chromatographic separation
of the analytes was achieved on a Discovery C18 (100 × 2.1 mm I.D., particle size 3.0 μm) column using 0.1% formic acid in water — acetonitrile (40: 60) as mobile phase
with a flow rate of 0.2 mL/min. Diazepam was used as the internal standard. The compounds were ionized in the electrospray
ionization source of the mass spectrometer and were detected by selected reaction ion monitoring of the transitions of m/z 310 → m/z 44.3 for fluoxetine, m/z 296 → m/z 134 for norfluoxetine and m/z 285 → m/z 193 for the internal standard. The method has low limit of detection (LOD) of 0.02 ng/mL and 0.03 ng/mL for fluoxetine and
norfluoxetine, respectively. The inter- and intra-run precision was measured to be below 5.3% (relative standard deviation)
for both fluoxetine and norfluoxetine. The developed method was successfully used to investigate plasma concentrations of
fluoxetine and norfluoxetine in the pharmacokinetic study of Chinese volunteers who received fluoxetine orally. |
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