Zhou Chen-Jian, Liu Ya-Nan, Wang Anzhou, Wu Hualu, Xu Ren-Ai, Zhang Qiang
Wenzhou Central Hospital, Zhejiang, China.
The First Affiliated Hospital of Wenzhou Medical University, Zhejiang, China.
Heliyon. 2024 May 30;10(11):e32187. doi: 10.1016/j.heliyon.2024.e32187. eCollection 2024 Jun 15.
PAXLOVID™ (Co-packaging of Nirmatrelvir with Ritonavir) has been approved for the treatment of Coronavirus Disease 2019 (COVID-19). The goal of the experiment was to create an accurate and straightforward analytical method using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to simultaneously quantify nirmatrelvir and ritonavir in rat plasma, and to investigate the pharmacokinetic profiles of these drugs in rats. After protein precipitation using acetonitrile, nirmatrelvir, ritonavir, and the internal standard (IS) lopinavir were separated using ultra performance liquid chromatography (UPLC). This separation was achieved with a mobile phase composed of acetonitrile and an aqueous solution of 0.1% formic acid, using a reversed-phase column with a binary gradient elution. Using multiple reaction monitoring (MRM) technology, the analytes were detected in the positive electrospray ionization mode. Favorable linearity was observed in the calibration range of 2.0-10000 ng/mL for nirmatrelvir and 1.0-5000 ng/mL for ritonavir, respectively, within plasma samples. The lower limits of quantification (LLOQ) attained were 2.0 ng/mL for nirmatrelvir and 1.0 ng/mL for ritonavir, respectively. Both drugs demonstrated inter-day and intra-day precision below 15%, with accuracies ranging from -7.6% to 13.2%. Analytes were extracted with recoveries higher than 90.7% and without significant matrix effects. Likewise, the stability was found to meet the requirements of the analytical method under different conditions. This UPLC-MS/MS method, characterized by enabling accurate and precise quantification of nirmatrelvir and ritonavir in plasma, was effectively utilized for pharmacokinetic studies in rats.
帕罗韦德™（奈玛特韦与利托那韦的复方制剂）已被批准用于治疗2019冠状病毒病（COVID-19）。该实验的目的是建立一种准确、简便的超高效液相色谱串联质谱法（UPLC-MS/MS），用于同时定量大鼠血浆中的奈玛特韦和利托那韦，并研究这些药物在大鼠体内的药代动力学特征。使用乙腈进行蛋白沉淀后，通过超高效液相色谱（UPLC）分离奈玛特韦、利托那韦和内标（IS）洛匹那韦。使用由乙腈和0.1%甲酸水溶液组成的流动相，通过反相柱二元梯度洗脱实现分离。采用多反应监测（MRM）技术，在正电喷雾电离模式下检测分析物。在血浆样品中，奈玛特韦的校准范围为2.0 - 10000 ng/mL，利托那韦的校准范围为1.0 - 5000 ng/mL，均呈现良好的线性。奈玛特韦和利托那韦的定量下限（LLOQ）分别为2.0 ng/mL和1.0 ng/mL。两种药物的日间和日内精密度均低于15%，准确度范围为 - 7.6%至13.2%。分析物的提取回收率高于90.7%，且无明显基质效应。同样，在不同条件下稳定性均符合分析方法要求。这种UPLC-MS/MS方法能够准确、精确地定量血浆中的奈玛特韦和利托那韦，有效地用于大鼠的药代动力学研究。
