Determination of indometacin in whole-blood microsamples using HPLC with electrochemical detection [Determinacion de indometacina en micromuestras de sangre empleando clar con deteccion electroquimica]
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abstract
A new high performance liquid chromatographic method with electrochemical detection was developed to evaluate indomethacin whole blood levels in microsamples coming from small species. The assay was performed after a single step of protein precipitation with acetonitrile for sample preparation. Indomethacin and naproxen (internal standar) were separated on a reversed-phase column Nova Pak C18 using a mixture of methanol and 75 mM ammonium phosphate buffer (62:38), v/v, pH = 3) as mobile phase. For the electrochemical detection, an oxidation potential of %2b1.1 V was set on the working electrode vs. Ag/AgCl. The detection limit of the assay was 25 ng-ml. Total run-time was lesser than 5 minutes. The method was linear within the range of 0.25 μg/ml, with coefficients of variation lower than 13%25. The assay demonstrated to be suitable for routine drug laboratory work, but it can also be used for clinical monitoring of indomethacin in small volume-limited samples.
