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Determination of flomoxef in human plasma by liquid chromatography/electrospray ionization tandem mass spectrometry
Authors:Oxana Yu. Kravtsova  Sergey A. Paramonov  Natalya I. Vasilevich  Denis N. Kazyulkin  Ekaterina Vlasova  Michael Engsig
Affiliation:1. Bioscreening Laboratory Co. Ltd, , Moscow, 125480 Russia;2. Russian Innovation Center Nycomed: a Takeda Company ‘Fusion Park’ Business‐Center 2, , Moscow, 119048 Russia
Abstract:
A specific, sensitive, rapid and reproducible method for the determination of flomoxef in human plasma using high‐performance liquid chromatography–tandem mass spectrometry was developed and validated. Flomoxef was detected using an electrospay ionization method operated in negative‐ion mode. Chromatographic separation was performed in gradient elution mode on a Luna® C18(2) column (3 μm , 20 × 4.0 mm) at a flow rate of 1 mL/min and runtime 3.5 min. The mobile phase consisted of acetonitrile and water containing 0.1% formic acid as additive. Extraction of flomoxef from plasma and precipitation of plasma proteins was performed with acetonitrile with an absolute recovery of 86.4 ± 1.6%. The calibration curve was linear with a correlation coefficient of 0.999 over the concentration range 10–5000 ng/mL and the lower limit of quantification was 10 ng/mL. The intra‐ and inter‐day precisions were <11.8%, while the accuracy ranged from 99.6 to 109.0%. A stability study of flomoxef revealed that it could be successfully analyzed at 4ºС over 24 h, but it was unstable in solutions at room temperature during short‐term storage (4 h) and several freeze–thaw cycles. Copyright © 2013 John Wiley & Sons, Ltd.
Keywords:flomoxef  human plasma  LC‐MS/MS analysis  tandem mass‐spectrometry
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