Determination of carbenicillin epimers in plasma and urine with high-performance liquid chromatography.

A high-performance liquid chromatographic method was developed for determining the concentrations of carbenicillin (CBPC) epimers in plasma and urine. Samples were prepared for HPLC analysis by solid-phase extraction and the concentrations of CBPC epimers were determined using reversed-phase HPLC with a mixture of 0.05 M ammonium acetate and methanol as a mobile phase. Baseline separation of the two epimers was observed for both plasma and urine samples with a detection limit of ca. 10 micrograms/ml. No peaks interfering with either of the CBPC epimers were observed on the HPLC chromatograms for blank plasma and urine. The presented method was used to determine the protein binding of CBPC epimers in vitro in human and rabbit plasma. The stereoselectivity of the binding of CBPC appeared to be reversed in human and rabbit plasma.