RP-HPLC测定人血浆中伏立康唑浓度及其应用分析  

Determination of voriconazole concentration in human plasma by RP-HPLC and analysis of its application

作　　者：吴水发 林志强[1] 洪丽绵 张清泉[1] 陈喆鸣 陈婷婷[1] WU Shuifa;LIN Zhiqiang;HONG Limian;ZHANG Qingquan;CHEN Zheming;CHEN Tingting(Department of Pharmacy,Quanzhou First Hospital Affiliated to Fujian Medical University,Fujian,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属泉州第一医院药剂科,福建泉州362000

出　　处：《中国医药科学》2021年第17期70-75,共6页China Medicine And Pharmacy

基　　金：福建省泉州市科技计划项目(2018N061S);福建医科大学启航基金项目(2018QH1187)。

摘　　要：目的建立并验证反相高效液相色谱法(RP-HPLC法)测定人血浆中伏立康唑浓度的方法学。方法先后采用乙腈蛋白沉淀法、乙酸乙酯-正己烷(4∶1,V/V)萃取法、氮气吹干法、少量流动相稀释法处理血浆样本。以Shim-pack Scepter C_(18-120)(250 mm×4.6 mm,5μm)为色谱柱,卡马西平为内标物,甲醇-水(57∶43,V/V)为流动相,进行等度洗脱,柱温为38℃,流速1.1 ml/min,检测波长为255 nm。考察该方法学的专属性、精密度、准确度、稳定性、实用性。结果血浆样本预处理后,伏立康唑和卡马西平有效浓集,内源性物质干扰小。伏立康唑血药浓度在0.03125~16μg/ml具有良好的线性关系,即y=0.3791x+0.0039(R^(2)=0.9992,N=10),提取回收率大于92%,日内、日间精密度考察均显示RSD小于10%,各项稳定性考察均显示RSD小于15%。2020年共检测230个样本,中位数为2.19(1.14~3.50)μg/ml,其中60.87%在目标范围内,21.74%低于目标值,17.39%高于目标值。结论本方法学线性好,灵敏度高,专属性强,准确且可靠,重现性好,可用于伏立康唑的治疗药物监测。Objective To establish and validate the methodology for the determination of voriconazole concentration in human plasma by reversed-phase high performance liquid chromatography(RP-HPLC method).Methods The plasma samples were treated by acetonitrile protein precipitation,ethyl acetate-hexane(4∶1,V/V)extraction,nitrogen blow dry method and small amount of mobile phase dilution method.The gradient elution was performed on a Shim-pack Scepter C_(18-120)(250 mm×4.6 mm,5μm)column with carbamazepine as the internal standard and methanol-water(57∶43,V/V)as the mobile phase,with column temperature of 38℃,flow rate of 1.1 ml/min and detection wavelength of 255 nm.The specificity,precision,accuracy,stability and practicality of the methodology were investigated.Results After pretreatment of plasma samples,voriconazole and carbamazepine were effectively concentrated with little interference from endogenous substances.The blood concentration of voriconazole had good linearity in the range of 0.03125-16μg/ml,i.e.,y=0.3791x+0.0039(R^(2)=0.9992,N=10),the extraction recovery was greater than 92%.Both intra-day and inter-day precision examination showed that the relative standard deviation(RSD)was less than 10%,and each stability examination showed that the RSD was less than 15%.A total of 230 samples were tested in 2020,with the median value of 2.19(1.14-3.50)μg/ml,of which 60.87%were within the target range,21.74%were below the target value and 17.39%were above the target value.Conclusion The method is linear,sensitive,specific,accurate and reliable with good reproducibility,so it can be useful for the therapeutic drug monitoring of voriconazole.

关 键 词：反相高效液相色谱法 伏立康唑 卡马西平 治疗药物监测 

分 类 号：R969.1[医药卫生—药理学]