Romanian Society of Pharmaceutical Sciences
 

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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

1.Department of Analytical Chemistry, Faculty of Pharmacy, “Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania
2 .Department of Toxicology, Faculty of Pharmacy, “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania
3 .Department of Pharmaceutical Technology and Biopharmacy, Faculty of Pharmacy, “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania
4 .Department of Paediatrics, Faculty of Medicine, “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania

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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.