Determination of amikacin in microlitre quantities of biological fluids by high-performance liquid chromatography using 1-fluoro-2,4-dinitrobenzene derivatization.

Pre-column derivatization of amikacin with 1-fluoro-2,4-dinitrobenzene in 25 microliter of guinea pig plasma or human serum produced a stable chromophore which was measured by UV detection after rapid separation on normal-phase or reversed-phase high-performance liquid chromatography systems. The reversed-phase system, selected for routine analysis due to instability of the normal-phase column, consisted of an Ultrasphere-ODS C18 column preceded by a guard column, and used acetonitrile--water (68:32) as the mobile phase. A high degree of linearity was found in the range of 2-64 microgram/ml with a coefficient of variation averaging less than 5%.