Wang Yingying, Gu Er-Min, Du Xiaoxiang, Xu Ren-Ai, Lin Guanyang
Department of pharmacy, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
The First People's Hospital of Jiashan, Jiaxing, China.
Front Pharmacol. 2021 May 20;12:641872. doi: 10.3389/fphar.2021.641872. eCollection 2021.
The contribution of the metabolites of linezolid to the associated myelosuppression is unknown in patients who are renal impairment. In this research, the purpose of our experiment was to explore and develop a quick and robust ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay for the determination of linezolid and its metabolite PNU-142300 in human serum simultaneously. The analytes were prepared using a simple and convenient approach with acetonitrile for protein crash, and then separated from the matrix on a Waters Acquity Ultra performance liquid chromatography (UPLC) BEH C18 (2.1 mm × 50 mm, 1.7 μm) column in a program of gradient elution, where the mobile phase was consisted of water with 0.1% formic acid and acetonitrile, and was placed at 0.40 ml/min flow rate. Multiple reaction monitoring (MRM) was employed and conducted for UPLC-MS/MS detection with ion transitions at 338.01 → 296.03 for linezolid, 369.96 → 327.98 for PNU-142300 and 370.98 → 342.99 for tedizolid (Internal standard, IS), respectively. This method had good linearity respectively in the calibration range of 0.01-20 μg/ml for linezolid, and 0.05-100 μg/ml for PNU-142300. In the intra- and inter-day, the precision of linezolid and PNU-142300 was below 14.2%, and the accuracy in this method was determined to be from -9.7 to 12.8%. In addition, recovery and matrix effect of the analytes were all found to be acceptable, and the analytes during the assay and storage in serum samples were observed to be stable. The novel optimized UPLC-MS/MS assay was also successfully employed to determine the concentration levels of linezolid and PNU-142300 in human serum. The results showed that linezolid-associated myelosuppression occurs more frequently in patients with renal insufficiency, and the metabolite-to-parent concentration ratio of PNU-142300 is predicted to reduce this toxicity of myelosuppression.
对于肾功能损害患者，利奈唑胺代谢产物对相关骨髓抑制的作用尚不清楚。在本研究中，我们实验的目的是探索并开发一种快速且可靠的超高效液相色谱串联质谱（UPLC-MS/MS）分析法，用于同时测定人血清中的利奈唑胺及其代谢产物PNU-142300。分析物采用简单便捷的方法制备，用乙腈进行蛋白沉淀，然后在Waters Acquity超高效液相色谱（UPLC）BEH C18（2.1 mm×50 mm，1.7μm）柱上通过梯度洗脱程序与基质分离，流动相由含0.1%甲酸的水和乙腈组成，流速设定为0.40 ml/min。采用多反应监测（MRM）进行UPLC-MS/MS检测，利奈唑胺的离子跃迁为338.01→296.03，PNU-142300的离子跃迁为369.96→327.98，替加环素（内标，IS）的离子跃迁为370.98→342.99。该方法在利奈唑胺0.01 - 20μg/ml和PNU-142300 0.05 - 100μg/ml的校准范围内分别具有良好的线性。在日内和日间，利奈唑胺和PNU-142300的精密度均低于14.2%，该方法的准确度测定为-9.7%至12.8%。此外，分析物的回收率和基质效应均被认为是可接受的，并且观察到分析物在血清样品的测定和储存过程中是稳定的。新优化的UPLC-MS/MS分析法也成功用于测定人血清中利奈唑胺和PNU-142300的浓度水平。结果表明，利奈唑胺相关的骨髓抑制在肾功能不全患者中更频繁发生，并且预计PNU-142300的代谢产物与母体浓度比可降低这种骨髓抑制毒性。
