摘要
目的 建立测定人血浆中伏立康唑浓度的高效液相色谱(HPLC)法.方法 采用乙腈蛋白沉淀法对血液样本进行处理.色谱柱为 Shimadzu InertSustain C18 柱(250mm×4.6mm,5μm),流动相为甲醇-乙腈-磷酸盐缓冲液(33∶28∶39,V/V/V),流速为1.0 mL/min,柱温为 30℃,检测波长为 262 nm,进样量为 20 μL.结果 伏立康唑的质量浓度在 0.1～8 μg/mL范围内与伏立康唑和酮康唑峰面积的比值线性关系良好(R2=0.999 8,n=8);定量限为 0.07 μg/mL.低、中、高质量浓度(0.125,1,8 μg/mL)样品的日内及日间精密度、冻融及长期稳定性试验结果的RSD均小于 5%;提取回收率分别为(97.32±1.71)%,(97.61±1.22)%,(96.38±0.84)%.60 例确诊为侵袭性真菌感染患者的血浆中,伏立康唑的质量浓度为 0.278～9.129 μg/mL.结论 所建立的方法简单、快速、稳定性良好,适用于伏立康唑的血药浓度监测.
Abstract
Objective To establish a high-performance liquid chromatography(HPLC)method for the determination of the concentration of voriconazole in human plasma.Methods Acetonitrile protein precipitation method was used to process blood samples.The chromatographic column was InsertSustain C18 column(250 mm×4.6 mm,5 μm),the mobile phase was methanol-acetonitrile-phosphate buffer(33∶28∶39,V/V/V),the flow rate was 1.0 mL/min,the column temperature was 30℃,the detection wavelength was 262 nm,and the injection volume was 20 μL.Results The linear range of voriconazole was 0.1-8 μg/mL(R2=0.999 8,n=8).The limit of quantification was 0.07 μg/mL.The RSDs of the intra-day and inter-day precision,freeze-thaw,and long-term stability test results for samples with low-,medium-,and high-concentrations of 0.125,1,8 μg/mL were all lower than 5% .The extraction recovery rates were(97.32±1.71)%,(97.61±1.22)%,and(96.38±0.84)%,respectively.In the plasma of 60 patients diagnosed with invasive fungal infections,the concentration of voriconazole was in the range of 0.278-9.129 μg/mL.Conclusion The established method is simple,rapid,and has good specificity,which is suitable for monitoring the blood concentration of voriconazole.
基金项目
四川省南充市科学技术和知识产权局首批重点实验室建设项目(NCKL201711)
川北医学院校级科研发展计划项目(CBY17-A-YB55)
国家科技期刊平台
NSTL
万方数据