Zhao Tinghui, Liu Lu, Yang Guangjie, Yu Hengyi, Qiu Lihui, Li Xiping, Xiang Dong, Gong Xuepeng
Department of Pharmacy, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Department of Pharmacy, Wuhan Mental Health Center, Wuhan, Hubei 430030, China.
J Mass Spectrom Adv Clin Lab. 2025 Jun 1;37:39-48. doi: 10.1016/j.jmsacl.2025.05.001. eCollection 2025 Aug.
Colistin serves as the last line of defense against multidrug-resistant Gram-negative bacterial infections and is commonly administered in clinical practice as its prodrug, colistin methanesulfonate (CMS). However, due to its notable nephrotoxicity and narrow therapeutic window, therapeutic drug monitoring (TDM) is essential.
To develop an optimal LC-MS/MS method for the quantification of colistin and CMS in human plasma and to apply it to stability studies and TDM.
Colistin A, colistin B, and internal standard (IS, polymyxin B2) were extracted from plasma using solid phase extraction columns. Sample separation was performed using a Welch Ultimate LP-C18 column with a 5-minute gradient elution consisting of water and acetonitrile, both supplied with 1.0% formic acid. The CMS concentration was obtained by comparing the total amount of colistin in acid-hydrolyzed and non-acid-hydrolyzed plasma.
Colistin A and colistin B showed excellent linearity in the concentration range of 0.1-10.0 μg/mL (R > 0.995) with acceptable specificity, accuracy (90.97 %-114.65 %), precision (RSD < 15 %), matrix effect (RSD < 15 %), and recovery (91.93 %-100.93 %). CMS in five commonly used clinical infusion solutions was stable when stored at room temperature for 8 h or at 4 °C for 24 h. The whole blood and plasma samples of CMS are susceptible to degradation at room temperature but are stable on ice. Plasma concentrations of colistin and CMS were accurately determined in three critically ill patients.
The method we have developed is robust and streamlined, and has successfully demonstrated the potential feasibility for future TDM applications of colistin and CMS in critically ill patients.
黏菌素是对抗多重耐药革兰氏阴性菌感染的最后一道防线，在临床实践中通常以其前体药物黏菌素甲磺酸盐（CMS）的形式给药。然而，由于其显著的肾毒性和狭窄的治疗窗，治疗药物监测（TDM）至关重要。
开发一种用于定量测定人血浆中黏菌素和CMS的最佳液相色谱-串联质谱（LC-MS/MS）方法，并将其应用于稳定性研究和TDM。
使用固相萃取柱从血浆中提取黏菌素A、黏菌素B和内标（IS，多黏菌素B2）。采用Welch Ultimate LP-C18柱进行样品分离，以水和乙腈组成的流动相进行5分钟梯度洗脱，二者均含有1.0%甲酸。通过比较酸水解和非酸水解血浆中黏菌素的总量来获得CMS浓度。
黏菌素A和黏菌素B在0.1 - 10.0 μg/mL浓度范围内表现出良好的线性（R > 0.995），具有可接受的特异性、准确度（90.97% - 114.65%）、精密度（RSD < 15%）、基质效应（RSD < 15%）和回收率（91.93% - 100.93%）。五种常用临床输液溶液中的CMS在室温下储存8小时或4°C下储存24小时时稳定。CMS的全血和血浆样品在室温下易降解，但在冰上稳定。在三名重症患者中准确测定了黏菌素和CMS的血浆浓度。
我们开发的方法稳健且精简，并已成功证明黏菌素和CMS在重症患者中进行未来TDM应用的潜在可行性。
