摘要
目的建立全自动二维高效色谱法(2D-HPLC)测定血清中伏立康唑的浓度并应用于临床。方法第一维色谱柱为ASTON SC2柱(4.6mm×25mm,5μm),流动相为20mmol/L磷酸铵∶乙腈∶甲醇=3∶1∶1(V∶V∶V,磷酸调pH=5.4),流速1.2mL/min;第二维色谱柱为SHIMADZU C18柱(4.6 mm×150 mm,5μm),流动相为10mmol/L醋酸铵(三乙胺调pH7.0)∶10mmol/L醋酸铵(三氟乙酸调pH3.0)∶甲醇∶乙腈=30∶10∶10∶50(V∶V∶V∶V),流速1.0mL/min;中间柱为ASTON SH柱(3mm×10mm,5μm),辅助流动相为纯水。样品由一维色谱柱萃取,经中间柱捕获并转移至第二维色谱柱分析,检测波长256nm,柱温40℃,进样量200μL。结果伏立康唑在0.28～22.56μg/mL范围内线性关系良好,r=0.999 9,最低定量限为0.28μg/mL,方法回收率为99.01%～104.62%,提取回收率为85.81%～89.82%,日内、日间精密度RSD均小于9%。结论该方法操作简单、准确、灵敏,符合血清样品的测定要求,适用于伏立康唑临床血药浓度测定。
Objective To establish the two-dimensional high performance liquid chromatography (2D-HPLC) for the determination of voriconazole in human serum and its application in clinical practice. Methods The first-dimension chromatographic column was the ASTON SC2 column (3.5 mm×25 mm, 5 μm) 20 mmol/L ammonium phosphate-acetonitrile-methanol(3 ∶1 ∶1, V ∶V ∶V, phosphoric acid adjusted pH = 5.4), and the flow rate of 1.2 mL/min . The second-dimension chromatographic column was the SHIMADZU C18 column (4.6 mm×150 mm, 5 μm) was 10 mmol/L ammonium acetate (triethylamine adjusted to pH = 7.0)– 10 mmol/L ammonium acetate (trifluoroacetic acid adjusted pH = 3.0)-Methanol- Acetonitrile(30 ∶10 ∶10 ∶50, V ∶V ∶V ∶V)and the flow rate of 1.0 mL/min. The middle chromatographic column was the ASTON SH column (3 mm×10 mm, 5 μm ) with pure water as the auxiliary mobile phase. The sample was extracted by the first-dimension chromatographic column, captured by the middle chromatographic column and transferred to the second-dimension chromatographic column for analysis. The detection wavelength was 256 nm, the temperature was 40 ℃ and the injection volume was 200 μL. Results The linear relationship of voriconazole was good in the range of 0.28~22.56 ug/mL ( r =0.9999) and the minimum quantitative limit was 0.28 ug/mL. The recovery rate of the method was from 99.01% to 104.62%, the extraction recovery rate was from 85.81% to 89.82%, and RSD of inter-day and intra-day assays was less than 9%. Conclusion The method was simple, accurate and sensitive, and it meets the requirements of serum sample determination. Therefore, it is suitable for the clinical determination of voriconazole concentration in blood.
作者
王新茗
张仲林
袁明勇
Wang Xinming;Zhang Zhonglin;Yuan Mingyong(School of Pharmacy,Chengdu Medical College,Chengdu 610500,China;Department of Pharmacy,The First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)
出处
《成都医学院学报》
CAS
2019年第2期169-173,共5页
Journal of Chengdu Medical College
基金
四川省科技厅项目(No:2012JY0035
No:2014JY0019)
关键词
二维高效液相色谱
伏立康唑
血药浓度
Two-dimensional high performance liquid chromatography
Voriconazole
Blood concentration
