超高效液相色谱-串联质谱法测定奈玛特韦及利托那韦血药浓度  

Monitoring Concentration of Nirmatrelvir and Ritonavir by Ultra High Performance Liquid Chromatography-tandem Mass Spectormetry Method

作　　者：范菁 唐昊翔[3] 王银辉 范炜斌 谢姣 林彬[1,2] FAN Jing;TANG Haoxiang;WANG Yinghui;FAN Weibin;XIE Jiao;LIN Bin(Department of Pharmacy,the People's Hospital of Changxing County,Changxing 313100,China;Key Laboratory of Intelligent Pharmacy and Individualized Therapy of Huzhou,Changxing 313100,China;Center for Disease Control and Prevention of Changxing County,Changxing 313100,China;Department of Pharmacy,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China)

机构地区：[1]浙江省长兴县人民医院药学部,长兴313100 [2]湖州市智能药学与个体化治疗重点实验室,长兴313100 [3]浙江省长兴县疾病预防控制中心,长兴313100 [4]西安交通大学附属第二医院药学部,西安710004

出　　处：《医药导报》2024年第2期190-195,共6页Herald of Medicine

基　　金：浙江省医学会神经科学科研专项(2021ZYC-A19)。

摘　　要：目的建立一种具有高灵敏度、高稳定性并且通用性强的能同时测定人血浆中奈玛特韦和利托那韦血药浓度的超高效液相色谱-串联质谱法。方法分离色谱柱采用ACQUITY UPLC BEH C_(18)柱(2.1 mm×50 mm,1.7μm),采用梯度洗脱,流动相为100%乙腈-0.1%甲酸,流速为0.3 mL·min^(-1),柱温为45℃,进样量为2μL。以电喷雾离子(ESI+)作为离子源,多反应监测模式扫描(奈玛特韦m/z 500.20→319.10,奈玛特韦-D_(9)m/z 508.59→328.10,利托那韦m/z 721.30→426.10,^(13)C,^(2)H_(3-)利托那韦m/z 725.30→426.10)。选择2023年1月于长兴县人民医院接受奈玛特韦/利托那韦治疗的新型冠状病毒感染患者30例,测定其用药3 d后的奈玛特韦和利托那韦稳态谷浓度。结果人血浆中奈玛特韦及利托那韦的线性范围分别为0.10~10.00(R^(2)=0.9972)和0.05~5.00μg·mL^(-1)(R^(2)=0.9952)。奈玛特韦和利托那韦的回收率均>90%,批内以及批间精密度的相对标准差(RSD)均<10%。另外,奈玛特韦和利托那韦回收率范围为91.5%~97.0%,基质效应范围为92.4%~97.7%。临床结果表明新型冠状病毒感染患者奈玛特韦和利托那韦血药浓度个体差异性很大。结论该研究建立的同时测定人血浆中奈玛特韦和利托那韦浓度的检测方法操作方便、专属性强、准确度及精密度高,适用于临床患者奈玛特韦和利托那韦血药浓度监测。Objective To establish a highly sensitive,stable,and universally applicable ultra-high-performance liquid mass spectrometry tandem method(UPLC-MS/MS)for simultaneous determination of nirmatrelvir and ritonavir blood concentrations in human plasma.Methods The separation was performed on an ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm,1.7μm)with gradient elution,and the mobile phase consisted of 0.1%formic acid-water and 100%acetonitrile at the flow rate of 0.3 mL·min^(-1).The column temperature was 45℃,and the injection volume was 2μL.Electrospray ionization as ion source(ESI+)was used as the ion source and multiple reactions monitoring mode(nirmatrelvir m/z 500.20→319.10,nirmatrelvir-D_(9) m/z 508.59→328.10,ritonavir m/z 721.30→426.10,^(13)C,^(2)H_(3-)ritonavir m/z 725.30→426.10)was adopted.Thirty patients with coronavirus disease 2019(COVID-19)treated with nirmatrelvir and ritonavir at the People's Hospital of Changxing County in Jan.2023 were selected to measure their steady-state trough concentrations of nirmatrelvir and ritonavir after 3 days of treatment.Results The linear range of nirmatrelvir was 0.100-10.0μg·mL^(-1)(R^(2)=0.9972),and the linear range of nirmatrelvir was 0.050-5.00μg·mL^(-1)(R^(2)=0.9952).The recovery rates of nirmatrelvir and lopinavir were both>90%and the intra-batch and inter-batch precision relative standard deviations(RSDs)were both<10%.Additionally,the recovery ranges for nirmatrelvir and lopinavir were 91.5%-97.0%,and the matrix effects ranged from 92.4%to 97.7%.The results of clinical samples showed that the plasma concentrations of nirmatrelvir and ritonavir in patients with COVID-19 varied greatly among individuals.Conclusion The method for simultaneous determination of nirmatrelvir and ritonavir concentrations in human plasma established in this study is convenient,highly specific,highly accurate,with high precision,which is suitable for monitoring the concentrations of nirmatrelvir and ritonavir in patients.

关 键 词：奈玛特韦 利托那韦 超高效液相色谱-串联质谱法 新型冠状病毒 

分 类 号：R978.7[医药卫生—药品] R969.1[医药卫生—药学]