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

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 39; no. 5; pp. 996 - 1005
• Main Authors: Isla, A., Arzuaga, A., Maynar, J., Gascón, A.R., Solinís, M.A., Corral, E., Pedraz, J.L.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 04-10-2005; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Anti-Bacterial Agents - blood; Biological and medical sciences; Calibration; Cefepime; Ceftazidime; Ceftazidime - blood; Cephalosporins - blood; Chromatography, High Pressure Liquid; Continuous hemodiafiltration; Fundamental and applied biological sciences. Psychology; General pharmacology; Hemofiltration; HPLC; Medical sciences; Pharmacology. Drug treatments; Quality Control; Reference Standards; Reproducibility of Results; Stability; Stability; Cefepime; Ceftazidime; HPLC; Continuous hemodiafiltration; Human; Biological fluid; β-Lactams; HPLC chromatography; Ceftazidime: Cefepime: HPLC: Continuous hemodiafiltration: Stability; Blood plasma; Cephalosporin derivatives; Antibiotic; Antibacterial agent; Physicochemical properties; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2005.05.027

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.