Fast, fully automated analysis of voriconazole from serum by LC–LC–ESI-MS–MS with parallel column-switching technique

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 814; no. 2; pp. 361 - 367
• Main Authors: Egle, Hannes, Trittler, Rainer, König, Armin, Kümmerer, Klaus
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25.01.2005; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Antifungal Agents - blood; Automation; Biological and medical sciences; Calibration; Chromatography, Liquid - instrumentation; Chromatography, Liquid - methods; Column-switching; Fundamental and applied biological sciences. Psychology; General pharmacology; LC–LC–ESI-MS–MS; Medical sciences; On-line sample preparation; Pharmacology. Drug treatments; Pyrimidines - blood; Reference Standards; Serum; Spectrometry, Mass, Electrospray Ionization - methods; Triazoles - blood; Voriconazole; Serum; On-line sample preparation; Column-switching; LC–LC–ESI-MS–MS; Voriconazole; Biological fluid; Antifungal agent; Triazole derivatives; Column chromatography; Switching
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2004.10.069

Voriconazole is a novel broad-spectrum antifungal agent. We developed an on-line LC–LC–MS–MS method for fully automated and direct analysis of voriconazole in raw human serum. After injection of human serum size-selective sample fractionation and analyte extraction was achieved using an extraction column (25 mm × 4 mm) packed with a restricted access material (RAM, LiChrospher ® ADS C 8, 25 μm). On-line transfer of voriconazole from the extraction column was followed by chromatography separation on a C 18 column. Detection was done by ESI-MS–MS. The total analysis time was 13 min, managed by parallel extraction and chromatographic separation. This LC–MS assay was fully validated. The lower limit of quantification was 0.05 μg/ml. The automated inline extraction of voriconazole described here eliminates the need for difficult and time-consuming sample pre-treatment. Other advantages of the new method are that only a small quantity (5 μl) of serum is needed and that the method is very specific.