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.
VoriconazoleIsotope dilution liquid chromatography-tandem mass spectrometryReference method
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