摘要
目的 建立高效液相色谱法(HPLC)同时检测洛匹那韦(LPV)和利托那韦(RTV)血药浓度的方法.方法 流动相为乙腈和0.01 mol/L磷酸二氢钠,体积比56﹕44,流速1 ml/min,紫外检测波长为210 nm和215 nm双波长,柱温35℃.取血浆100 μl,加入2 μg/ml地西泮内标100 μl,涡旋振荡1 min,12 000 r/min离心10 min,取上清液100 μl进样分析.结果 双波长均显示LPV在1.57~201 μg/ml浓度范围内以及RTV在0.38~48.5 μg/ml浓度范围内线性均良好(分别为r=0.999 5和r=0.999 6).在LPV和RTV极低浓度(2.47 μg/ml、0.62 μg/ml)、低浓度(7.41 μg/ml、1.85 μg/ml)、中浓度(22.22 μg/ml、5.56 μg/ml)、高浓度(66.67 μg/ml、16.67 μg/ml)质控样品中,210 nm处LPV和RTV的日内变异和日间变异分别为0.78%~12.32%、2.07%~13.41%和2.70%~19.41%、4.36%~24.86%;215 nm处分别为 2.36%~9.30%、2.82%~11.20%和 1.18%~27.12%、1.30%~28.13%.四个浓度质控样品的准确度,210 nm条件下是85.30%~123.33%,215 nm条件下是89.20%~110.81%.四个浓度质控样品的方法学回收率,210 nm条件下是85.08%~111.66%,215 nm条件下是91.05%~111.41%.结论 本方法经济便捷,适用于临床LPV和RTV血药浓度的检测.
Objective To establish a method for the simultaneous determination of Lopinavir(LPV)and Ritonavir(RTV)blood concentrations by high performance liquid chromatography(HPLC).Methods The mobile phase was acetonitrile and 0.01 mol/L sodium dihydrogen phosphate,the volume ratio was 56:44,the flow rate was 1 ml/min,the UV detection wavelength was 210 nm and 215 nm,and the column temperature was 35℃.100μl of plasma was taken,100μl of 2μg/ml diazepam internal standard was added,vortex shaking for 1 min,centrifuged at 12000 r/min for 10 min,and 100μl of supernatant was taken into the sample for analysis.Results The linearity of both wavelengths was good at 1.57~201μg/ml LPV and 0.38~48.5μg/ml RTV(r=0.9995 and r=0.9996,respectively).In the quality control samples of LPV and RTV at extremely low concentration(2.47μg/ml,0.62μg/ml),low concentration(7.41μg/ml,1.85μg/ml),mediumconcentration(22.22μg/ml,5.56μg/ml)and high concentration(66.67μg/ml,16.67μg/ml),the intra-day variation and inter-day variation of LPV and RTV at 210 nm were 0.78%~12.32%,2.07%~13.41%and 2.70%~19.41%,4.36%~24.86%,respectively.At 215 nm,they were 2.36%~9.30%,2.82%~11.20%and 1.18%~27.12%,1.30%~28.13%,respectively.The accuracy of the four concentration quality control samples was 85.30%~123.33%at 210 nm,and 89.20%~110.81%at 215 nm.The methodological recovery of the four concentration quality control samples was 85.08%~111.66%at 210 nm,and 91.05%~111.41%at 215 nm.Conclusion This method is economical,convenient,suitable for determination of clinic LPV and RTV blood concentrations.
作者
厉琳琳
陈方亮
谢颖洁
LI Lin-lin;CHEN Fang-liang;XIE Ying-jie(Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University、Enze Hospital,Taizhou Enze Medical Center(Group),Taizhou 318050,China)
出处
《医师在线》
2024年第4期17-21,共5页
Journal of Doctors Online
基金
2020年台州市第二批社会发展科技计划项目(20ywb76)。
