HPLC-MS/MS法测定人血浆中奈玛特韦和利托那韦浓度  

Determination of nirmatrelvir and ritonavir in human plasma by HPLC-MS/MS method

作　　者：徐宗良 李盛建 钱跹 段虎 周瑾 张倩倩 董昕 赵亮[1,2] XU Zong-liang;LI Cheng-jian;QIAN Xian;DUAN Hu;ZHOU Jin;ZHANG Qian-qian;DONG Xin;ZHAO Liang(School of Medicine,Shanghai University,SHANGHAI 200444,China;Department of Pharmacy,Shanghai Baoshan Luodian Hospital,SHANGHAI 201908,China)

机构地区：[1]上海大学医学院,上海200444 [2]上海市宝山区罗店医院药剂科,上海201908

出　　处：《中国新药与临床杂志》2024年第1期57-61,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：上海市宝山区卫生健康委优青(育才)计划(BSWSYC-2023-14)。

摘　　要：目的建立高效液相色谱质谱联用(HPLC-MS/MS)法测定人血浆奈玛特韦和利托那韦的血药浓度,并应用于治疗药物监测。方法血浆样品经蛋白沉淀后测定,流动相为0.1%甲酸水溶液(A相)、乙腈(B相),采用等度洗脱,流速为0.3 m L·min^(-1),柱温为40℃,使用EC-C18色谱柱,正离子模式下进行多反应监测。结果用于定量分析的离子通道分别为m/z 500→110(奈玛特韦),m/z721.2→296.3(利托那韦),m/z 614.4→421(内标瑞德西韦)。人血浆中奈玛特韦在0.01~10μg·mL^(-1)、利托那韦在0.01~2.0μg·mL^(-1)浓度范围内线性关系良好,提取回收率分别为79.56%~89.91%、82.03%~89.56%,日间及日内的RSD均小于15%,并且稳定性良好。采用该方法检测了34例新型冠状病毒肺炎(COVID-19)患者口服Paxlovid后的血药浓度,血药浓度范围在1.04~18.0μg·mL^(-1)之间。结论本研究建立的HPLC-MS/MS方法检测快速,灵敏度高,准确性好,能应用于COVID-19患者服用Paxlovid后的血药浓度监测。AIM To establish a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for the determination of blood drug concentrations of nirmatrelvir and ritonavir in human plasma and apply it to therapeutic drug monitoring(TDM).METHODS Plasma samples were precipitated,and the mobile phase consisted of 0.1%formic acid aqueous solution(phase A)and acetonitrile(phase B).Isocratic elution was used with a flow rate of 0.3 mL·min^(-1),a column temperature of 40℃,and an EC-C18 chromatographic column.Multiple reaction monitoring was conducted in positive ion mode.RESULTS The ion channels used for quantitative analysis were m/z 500→110(nirmatrelvir),m/z 721.2→296.3(ritonavir),and m/z 614.4→421(internal standard remdesivir).In human plasma,nirmatrelvir showed good linearity in the range of 0.01 to 10μg·mL^(-1),while ritonavir exhibited good linearity in the range of 0.01 to 2.0μg·mL^(-1).The extraction recoveries were 79.56%to 89.91%for nirmatrelvir and 82.03%to 89.56%for ritonavir.The RSD values for intraday and inter-day precision were all less than 15%,and the stability was satisfactory.The developed method was applied to measure the blood drug concentrations of 34 patients after oral administration of Paxlovid,which ranged from 1.04 to 18.0μg·mL^(-1).CONCLUSION The HPLC-MS/MS method established in this study is rapid,highly sensitive,and accurate,and it can be applied to monitor the therapeutic drug concentration monitoring of COVID-19 patients after taking Paxlovid.

关 键 词：色谱法 高压液相 串联质谱法 奈玛特韦 利托那韦 血药浓度 

分 类 号：R927.2[医药卫生—药学]