Simultaneous determination of coumarin, 7-hydroxycoumarin and 7-hydroxycoumarin glucuronide in human serum and plasma by high-performance liquid chromatography

Journal of Chromatography. B, Biomedical Applications
D P Bogan, R O'Kennedy

Abstract

A HPLC method was developed for the determination of the metabolites of coumarin and 7-hydroxycoumarin in plasma and serum. Separation was based on gradient elution of 7-hydroxycoumarin glucuronide, 7-hydroxycoumarin, coumarin and finally 4-hydroxycoumarin (which is used as an internal standard). Standards, prepared in plasma or serum, and samples were treated with trichloroacetic acid, mixed and centrifuged. The supernatant was removed and analyzed by reversed-phase high-performance liquid chromatography on a C18 column. The limit of detection was 50 ng/ml for 7-hydroxycoumarin and 200 ng/ml for coumarin and 7-hydroxycoumarin glucuronide. The linear range was 0.5-100 micrograms/ml for each of the analytes. The percentage relative standard deviation about the mean measured concentrations were all below 10%. There was no statistical difference between the standard curves prepared in plasma or serum. The method developed was applied to the determination of each of the three compounds in serum, after the administration of 7-hydroxycoumarin, and in plasma after the administration of coumarin. The concentrations of total 7-hydroxycoumarin in the serum samples were also determined by another HPLC method and the results were compared....Continue Reading

Citations

Dec 8, 2007·European Journal of Clinical Pharmacology·Sara AsimusMichael Ashton
May 24, 2014·Biomedical Chromatography : BMC·Aura Rocío HernándezDiana Marcela Aragón
Apr 20, 2006·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Ragini Vuppugalla, Reza Mehvar
Feb 27, 2018·Journal of Investigative Medicine : the Official Publication of the American Federation for Clinical Research·Rasha Tarif HamzaMahmoud T Sallam

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