Quantification of voriconazole in human bronchoalveolar lavage fluid using high-performance liquid chromatography with fluorescence detection

The quantification of voriconazole concentration in lung epithelial lining fluid to facilitate the management of pulmonary fungal colonisation or aspergillosis is of increasing interest. An accurate and reproducible high-performance liquid chromatography method to quantify voriconazole in human bronchoalveolar lavage (BAL) fluid was developed and validated. BAL samples were concentrated by freeze-drying and reconstituted with water prior to deproteinisation. Separation was achieved with a C18 column employing fluorescence detection (excitation: 260 nm, emission: 370 nm). The calibration curves were linear from 2.5 to 500 ng/mL. The intra- and inter-day precisions were within 7%. Accuracies ranged from 102% to 107%. The clinical applicability was established by successful measurement of voriconazole concentrations in lung transplant recipients. The assay provides an alternative approach for those with negligible access to liquid chromatography–tandem mass spectrometry instrumentation.

► Validation of a HPLC-fluorescence method for the determination of VRC in human BAL samples.
► Freeze-drying step to concentrate BAL samples, attaining high sensitivity with LLOQ of 2.5 ng/mL.
► Successful application in a clinical study in immunocompromised lung transplant recipients, who are at risk of low VRC concentrations.