2021年12月,美国食品和药物管理局(FDA)批准了Paxlovid®作为COVID-19的抗病毒治疗的紧急授权。Paxlovid®由两片组成,尼玛特雷韦和利托那韦.在肾功能不全的情况下,剂量调整是必要的。本研究的目的是建立同时测定人血清中尼马特雷韦/利托那韦的LC-HRMS方法,用于治疗药物的监测。在25μL人血清样品中加入内标沙奎那韦,然后用甲醇沉淀样品。分析物在C18柱上通过梯度洗脱分离,使用0.1%甲酸-水和甲醇的流动相,在0.4毫升/分钟的流速。注射体积为2μL,分析时间为5min。分析物的测定通过电喷雾电离以正模式通过全质量监测进行。检测到的尼马特雷韦离子,利托那韦和沙喹那韦分别为m/z500.24792、721.32004和671.39155。尼马特雷韦的线性浓度范围为78.13-20000ngmL-1,利托那韦的线性浓度范围为15.63-4000ngmL-1(r2>0.9900)。准确度范围为87.45%~104.63%,日内和日间精度RSD<15%。尼马特雷韦的回收率为98.72%~109.83%,利托那韦的比例为95.41~112.36%。尼马特雷韦的基质效应为88.31~97.73%,利托那韦的比例为85.17~103.05%。该方法用于测量17例患者的尼马特雷韦/利托那韦的谷浓度。nirmatrelvir的谷浓度为1331.7-8352.5ng/mL,利托那韦的浓度为53.4-1325.5ngmL-1,个体差异较大。方法简单,敏感,具体,和可重复的,可用于监测尼马特雷韦/利托那韦在COVID-19患者中的血药浓度和药代动力学研究。
In December 2021, the U.S. Food and Drug Administration (FDA) granted emergency authorization for Paxlovid® as an antiviral treatment for COVID-19. Paxlovid® is composed of two tablets,
nirmatrelvir and ritonavir. Dose adjustment is necessary in cases of renal insufficiency. The aim of present study is to establish a LC-HRMS method for simultaneous determination of
nirmatrelvir/ritonavir in human serum for therapeutic drug monitoring. Internal standard saquinavir was added in 25 μL human serum samples, and then the samples were precipitated with methanol. The analytes were separated by gradient elution on a C18 column, using a mobile phase of 0.1 % formic acid-water and methanol, at a flow rate of 0.4 mL/min. The injection volume was 2 μL, and the analysis time was 5 min. The determination of the analytes was performed by electrospray ionization in positive mode by full mass monitoring. The detected ions of nirmatrelvir, ritonavir and saquinavir were m/z 500.24792, 721.32004 and 671.39155, respectively. The linear concentration range for
nirmatrelvir was 78.13-20000 ng mL-1, for ritonavir was 15.63-4000 ng mL-1 (r2>0.9900). The accuracy ranged from 87.45 %∼104.63 %, and the intra-day and inter-day precision RSD was <15 %. The recovery of nirmatrelvir ranged from 98.72 %∼109.83 %, and that of ritonavir was 95.41∼112.36 %. The matrix effect of
nirmatrelvir was 88.31∼97.73 %, and that of ritonavir was 85.17∼103.05 %. This method was used to measure the trough concentrations of
nirmatrelvir/ritonavir in 17 patients. The trough concentration of nirmatrelvir was 1331.7-8352.5 ng/mL, and that of ritonavir was 53.4-1325.5 ng mL-1, with large individual differences. The method is simple, sensitive, specific, and reproducible, and can be used for monitoring the blood concentration and pharmacokinetic study of nirmatrelvir/ritonavir in COVID-19 patients.