High-performance liquid chromatographic assays for the quantification of amikacin in human plasma and urine.

Amikacin, an aminoglycoside antibiotic, is frequently coadministered with penicillins and broad-spectrum cephalosporins to synergize the activity of these agents. Sensitive, selective and reproducible high-performance liquid chromatographic assays have been developed for the quantification of amikacin in plasma and urine collected from human subjects. The plasma method involves the ultrafiltration of plasma prior to derivatization. An aliquot of plasma ultrafiltrate or urine is mixed with dimethyl sulfoxide and tris(hydroxymethyl)aminoethane followed by derivatization of amikacin with 1-fluoro-2,4-dinitrobenzene at 58 degrees C for 30 min. The reaction mixture is then injected directly onto a reversed-phase C18 column preceded by a guard column. The column is eluted with a mobile phase containing acetonitrile and 2-methoxyethanol in 1% Tris buffer. Amikacin derivative is detected at 340 nm. The methods were applied for the analysis of amikacin in plasma and urine samples from volunteers receiving amikacin and cefepime, a fourth-generation cephalosporin, in a clinical pharmacokinetic drug interaction study.