The application of high-pressure liquid chromatography assays for cephalosporin serum concentrations is difficult in uremic patients because of interference from nondialyzable substances. We developed a high-pressure liquid chromatography method for determining the serum concentration of ceftizoxime in normal and uremic patients. The method involves protein precipitation with acetonitrile, followed by removal of the acetonitrile with dichloromethane. Separation was accomplished with a reverse-phase (C-18) column and a mobile phase of 13% acetonitrile and 2.8% acetic acid. UV detection at 310 nm was used to monitor the peaks. This assay produced a linear relationship between peak height ratio and ceftizoxime concentration from 1.5 to 100 micrograms/ml. Samples from 30 patients were assayed by this method and by a bioassay, with a good correlation of results (r = 0.9832). The method was applicable equally to normal and uremic serum samples.
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