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A simple and rapid high-performance liquid chromatographic method has been developed for determination in human plasma of isepamicin (ISP), an aminoglycoside antibiotic agent. After protein precipitation and clean-up procedure to remove lipophilic contaminants, ISP is derivatized pre-column with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate for fluorescence detection. Chromatographic separations are achieved using C(18) column and mobile phase consisting of 20 mM KH(2)PO(4) containing 8 mM triethylamine (pH 7.0) and acetonitrile (78/22, v/v). Amikacin was used as an internal standard. The calibration curve was linear over a concentration range of 0.5-50 microg/ml. The limit of quantification was 0.5 microg/ml. The intra- and inter-day variabilities of ISP were both less than 17.5%. Both derivatives were stable for at least a week at ambient condition. This assay procedure should have useful application in therapeutic drug monitoring of ISP.

biological and pharmaceutical bulletin

Determination of Isepamicin in Human Plasma by HPLC with Fluorescence Detection after Derivatization Using 6-Aminoquinolyl-N-hydroxysuccinimidyl-carbamate