Abstract:

Objective To establish a candidate reference measurement method for serum voriconazole by isotope dilution liquid chromatography-tandem mass spectrometry （ID-LC-MS/MS）. Methods The samples were pretreated by protein precipitation （PPT） with methanol as precipitator. Using voriconazole -2H3 as internal standard，an ID-LC-MS/MS for the determination of voriconazole in serum was established. The linearity，limit of quantification （LOQ），limit of determination （LOD），accuracy，precision，matrix effect and stability were evaluated. Results The linear range of serum voriconazole was 0.2-16.0 μg·mL^-1，the limit of quantification was 0.08 μg·mL^-1，and the limit of determination was 0.016 μg·mL^-1. The recoveries were 97.26%-98.48%. The within-run coefficient of variation （CV） was <1%，and the between-run CV was < 2%. The relative matrix effect ranged from -1.87% to 0.33%. The samples were placed at room temperature [（23±2） ℃] for 4，8 h and repeatedly frozen and thawed for 5 times at -20 ℃，and the samples after pre-treatment were placed at room temperature and automatic injector （8 ℃） for 4，8，12 and 24 h，respectively. The recoveries were 95.63% - 99.50%，and the determination results were stable. The relative extended uncertainty was ≤3.34%. Conclusions A candidate reference method for the determination of serum voriconazole based on ID-LC-MS/MS has been established，which can be used for standardization.

Key words: Voriconazole, Isotope dilution liquid chromatography-tandem mass spectrometry, Reference method