Determination of ceftazidime and cefepime in plasma and dialysate-ultrafiltrate from patients undergoing continuous veno-venous hemodiafiltration by HPLC

Abstract

We have developed and validated a new, rapid and reproducible HPLC method for the determination of cefepime and ceftazidime in plasma and dialysate-ultrafiltrate samples obtained from intensive care unit (ICU) patients undergoing continuous veno-venous hemodiafiltration (CVVHDF). The method for plasma samples involved protein precipitation with acetonitrile, followed by washing with dichloromethane to remove apolar lipophilic compounds. Dialysate-ultrafiltrate samples did not require any preparation. Separation was performed on a μBondapak C18 (30 cm × 3.9 mm × 10 μm) with UV detection. The mobile phase contained acetate buffer: ACN and was delivered at 2 ml/min. The coefficients of determination of the calibration curves were always ≥0.998 and R.S.D.% of the response factors <10%. The intra and inter-assay precision and accuracy of the quality controls (QC) and limit of quantification (LOQ) were satisfactory in all cases. Plasma and dialysate-ultrafiltrate samples were stable at −20 and −80 °C for 2 months and also after three freeze/thaw cycles. Dialysate-ultrafiltrate samples were stable in the chromatographic rack for 24 h at room temperature, but we recommend storing processed plasma samples at 4 °C until the analysis. The described method has proved to be useful to give accurate measurements of ceftazidime and cefepime in samples obtained from patients undergoing CVVHDF.