High-performance liquid chromatographic assay for cefepime in serum.

A simple, rapid, specific and sensitive high-performance liquid chromatographic method was developed for the determination of cefepime 1-[[(6R, 7R)-7-[2-(2-amino-4-thiazolyl) glyoxylamido]-2-carboxy-8-oxo-5-thia-1-azabicyclo- [4.2.0]oct-2-en-3-yl]methyl]-1-methylpyrrolidinium hydroxide, inner salt, 7(2)-(Z)-(O-methyloxime) in human serum. Separation was achieved on a reversed-phase Ultrasphere XL-ODS column (75 x 4.6 mm I.D.). The mobile phase was 7% acetonitrile in 20 mM ammonium acetate (pH 4). Cefepime eluted in the range of 1.8-2.2 min. Detection was by UV absorbance at 254 nm. The lower limit of quantitation of cefepime in plasma was 0.5 microgram/ml. The average absolute recovery was 106.2 +/- 2.1%. The linear range was from 0.1 to 50 micrograms/ml, with a correlation coefficient greater than 0.999. The within-day C.V.s for human samples were 4.9 and 2.3% for 1 and 50 micrograms/ml, respectively. The between-day C.V.s for human serum samples were 14.5, 7.4 and 6.7 for 1, 25 and 50 micrograms/ml, respectively. Cefepime was found to be unstable in serum at room temperature. For delayed assay, samples must be stored at -80 degrees C.