HIGH-THROUGHPUT HPLC METHOD FOR RAPID
QUANTIFICATION OF KETOPROFEN IN HUMAN PLASMA

JEAN NEGRU1, DANIELA-SAVETA POPA2*, LAURIAN VLASE3, DANA IACOB4, MARCELA ACHIM3, VASILE DORNEANU1
1Department of Analytical Chemistry, Faculty of Pharmacy, “Gr. T. Popa” University of Medicine and Pharmacy, Iasi,
Romania
2Department of Toxicology, Faculty of Pharmacy, “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca,
Romania
3Department of Pharmaceutical Technology and Biopharmacy, Faculty of Pharmacy, “Iuliu Haţieganu” University of
Medicine and Pharmacy, Cluj-Napoca, Romania
4Department of Paediatrics, Faculty of Medicine, “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca,
Romania
*corresponding author: dspopa@yahoo.fr
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Abstract:
A new high-throughput HPLC assay for the quantification of ketoprofen in human plasma was developed and validated. The
separation was performed on a Zorbax SB-C18 column under isocratic conditions using a 55:45 (v/v) mixture of acetonitrile
and 1% trifluoroacetic acid in water at 45ºC with a flow rate of 1.5 mL/min. The detection of ketoprofen was performed at
257 nm. The human plasma samples (0.2 mL) were deproteinized with methanol (0.6 mL) and aliquots of 20 μL from
supernatants obtained after centrifugation were directly injected into the chromatographic system. The method showed a
good linearity (r > 0.9997), precision (CV > 6.0 %) and accuracy (bias < 4.8 %) over the studied range of 153.2 - 19155
ng/mL plasma. The lower limit of quantification (LLOQ) was 153 ng/mL and the recovery was between 96.5 - 103.6 %. The
method is not expensive, it doesn’t require a long time for plasma sample preparation and has a run-time of 2.1 min for
instrument analysis (the retention time of ketoprofen was 1.7 min). The developed and validated high-throughput method is
very simple, rapid and efficient, with wide applications in pharmacokinetics and bioequivalence studies.






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