Determination of free concentration of voriconazole in human plasma by HFCF-UF combined with HPLC
Objective To establish a high performance liquid chromatography(HPLC)method for determining the free concentration of voriconazole in human plasma and to use it for clinical therapeutic drug monitoring(TDM).Methods Hollow fiber centrifugation ultrafiltration(HFCF-UF)pretreatment was used to obtain free voriconazole in plasma,and the content was determined by HPLC.Chromatographic column:Diamond C18 column(250.0 mm × 4.6 nm,5 μm),column temperature:30 ℃;mobile phase:water-acetonitrile=60∶40(v∶v)at a flow rate of 1.0 mL·min-1;wavelength:254 nm;injection volume:20 µL.The specificity,linear relationship,lower limit of quantification,precision,recovery rate,and stability of this method were investigated,and clinical plasma samples were determined.Results The linearity was good within the range of 0.10-10.00 μg·mL-1,and the standard curve equation was y=28 607x-35.93(r=0.999 1),and the lower limit of quantification was 0.10 μg·mL-1;the absolute recovery rate and relative recovery rate were 95.73％-97.76％and 96.64％-102.93％,respectively.The relative standard deriation of intra-day and inter-day precision were both less than 10％;the stability was good when plasma samples were repeatedly freeze-thawed three times or stored at-40 ℃ for 7 days,and at room temperature for 6 hours after sample processing.The free concentration of voriconazole in 37 plasma samples was 0.38-7.18 μg·mL-1,and plasma protein binding rates were(50.84±14.76)％,and there was a certain correlation between free concentration and total concentration.Conclusion The HFCF-UF combined with HPLC method for determining the free concentration of voriconazole in plasma is a simple,accurate,and applicable method for clinical TDM.
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