Validation of an assay for voriconazole in serum samples using liquid chromatography-tandem mass spectrometry

Brian G. Keevil, Sheila Newman, Stephen Lockhart, Susan J. Howard, Caroline B. Moore, David W. Denning

    Research output: Contribution to journalArticlepeer-review


    A simple and rapid liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the analysis of voriconazole has been developed. For comparison, serum voriconazole was measured using HPLC and bioassay. For the HPLC-MS/MS assay, samples were prepared in a deep-well block by adding 10 μL of serum to 40 μL of 0.1 M zinc sulfate solution. Proteins were precipitated by adding 100 μL acetonitrile containing ketoconazole as internal standard. After vigorous mixing and centrifugation, 3 μL of the supernatant was injected into the HPLC-MS/MS system. An HPLC system was used to elute a C 18 cartridge (2 mm x 4 mm) at 0.6 mL/min with a step gradient of 50% to 100% methanol containing 2 mM ammonium acetate and 0.1% (vol/vol) formic acid. The column was maintained at 55°C, and the retention times were voriconazole 1.50 minutes and ketoconazole 1.47 minutes. Cycle time was 3 minutes, injection to injection. The analytes were monitored using a tandem mass spectrometer operated in multiple reaction monitoring mode using the following transitions: voriconazole m/z 350.0 > 224.1 and ketoconazole m/z 531.1>489.1. Within- and between-batch CVs were
    Original languageEnglish
    Pages (from-to)650-657
    Number of pages7
    JournalTherapeutic Drug Monitoring
    Issue number6
    Publication statusPublished - Dec 2004


    • Aspergillosis
    • Tandem mass spectrometry
    • Voriconazole


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