摘要
目的建立测定人血浆中伏立康唑游离浓度的高效液相色谱(HPLC)法,并用于临床治疗药物监测(TDM)。方法通过中空纤维离心-超滤(HFCF-UF)法预处理得到血浆中游离伏立康唑后,用HPLC法测定含量。色谱柱:Dia⁃monsil C18柱(250.0 mm×4.6 nm,5μm),柱温:30℃;流动相:水-乙腈=60∶40(v∶v),流速:1.0 mL·min^(-1);波长:254 nm;进样量:20μL。对该方法的专属性、线性关系、定量下限、精密度、回收率和稳定性进行考察,并用于临床血浆样品的测定。结果本研究建立的测定方法专属性良好,血浆中伏立康唑的游离浓度在0.10~10.00μg·mL^(-1)呈线性关系,标准曲线方程为y=28607x-35.93(r=0.9991),定量下限为0.10μg·mL^(-1),绝对回收率和相对回收率分别为95.73%~97.76%和96.64%~102.93%,日内和日间精密度的相对标准偏差均小于10%;血浆样品反复冻融3次或-40℃下保存7 d,处理后样品室温放置6 h稳定性良好。37份血浆样品的伏立康唑游离浓度为0.38~7.18μg·mL^(-1),血浆蛋白结合率为(50.84±14.76)%,游离浓度与总浓度之间有一定的相关性。结论HFCF-UF联合HPLC测定血浆中伏立康唑游离浓度的方法简便准确,适用于临床TDM。
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,5μm),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:20μL.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 0.10-10.00μg·mL^(-1),and the standard curve equation was y=28607x-35.93(r=0.9991),and the lower limit of quantification was 0.10μg·mL^(-1);the absolute recovery rate and relative recovery rate were 95.73%-97.76%and 96.64%-102.93%,res pe ctively.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 0.38-7.18μg·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.
作者
张甜甜
李文利
武玺坤
张志清
ZHANG Tian-tian;LI Wen-li;WU Xi-kun;ZHANG Zhi-qing(Department of Pharmacy,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,Hebei Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2024年第17期2557-2561,共5页
The Chinese Journal of Clinical Pharmacology
关键词
伏立康唑
游离浓度
中空纤维离心-超滤法
高效液相色谱法
voriconazole
free concentration
hollow fiber centrifugation ultrafiltration
high performance liquid chro⁃matography
