Rapid High-Performance Liquid Chromatography (HPLC) and Bioassay with Extended Analytical Range for Measured of Voriconazol (VRC) Blood Levels

A. A. PASCUAL, T. CALANDRA, D. SANGLARD, S. BOLAY, V. NIETH, L. A. DECOSTERD, J. BILLE, O. MARCHETTI

Author address: 

CHUV, Lausanne, Switzerland

Abstract: 

Background: VRC is a new antifungal agent active against Candida, Aspergillus, and emerging fungi. Inter-individual variability in VRC pharmacokinetics may result in over or under-dosing, that might be an issue in patients with life-threatening infections or risk of toxicity. Available HPLC and bioassay methods are time-demanding (extraction >4 h, run time >20 min) and have narrow analytical range (0.6-5 mg/L). Objective: To develop a rapid HPLC method and a bioassay with an extended analytical range. Methods: HPLC: VRC extraction from plasma after acetonitrile precipitation (60 min), reverse phase separation with UV detection at 255 nm (12 min). Bioassay: C. albicans mutant DSY2621 (916;cdr1, 916;cdr2, 916;flu, 916;mdr1, 916;cna), MIC VRC 0.008 mg/L. VRC standards (0.2 to 25 mg/L), VRC quality controls (0.25, 1, 4 and 16 mg/L): validation according to international guidelines (Shah et al, 2000). Accuracy: measured/nominal value x 100. Precision: SD/mean of measured values x 100. Results: Reproducible standard curves were obtained with both methods (r ≥ 0.99). Mean VRC recovery from plasma vs water was 102.5% ± 5.9 for HPLC and 99.1% ± 0.5 for bioassay. VRC was stable (+/8722; 10% of nominal value) at 4 and 21°C during 7 d, at -80°C during 4 months, and after 4 freeze-thaw cycles. Intra and inter run validations with quality controls (mean % values ± SD) Comparison of VRC levels measured by HPLC and by bioassay in 58 clinical samples showed high correlation (r=0.97, p
2004

abstract No: 

A-1477-200

Full conference title: 

44th Interscience Conference on Antimicrobial Agents and Chemotherapy
    • ICAAC 44th