LC-MS/MS法同时测定碳青霉烯耐药革兰氏阴性菌感染患者血浆中头孢他啶阿维巴坦、多黏菌素B及替加环素浓度

Quantitative determination of ceftazidime-avibactam, polymyxin B, and tigecycline in plasma of patients with carbapenem-resistant organism infection by LC-MS/MS

关键词:  头孢他啶阿维巴坦  多黏菌素B  替加环素  治疗药物监测  液相色谱串联质谱

Key words:  ceftazidime-avibactam  polymyxin B  tigecycline  therapeutic drug monitoring  LC-MS/MS

摘要:

目的：建立液相色谱串联质谱（LC-MS/MS）方法同时测定人血浆中头孢他啶阿维巴坦、多黏菌素B及替加环素3种药物共5种成分（头孢他啶、阿维巴坦、多黏菌素B1、B2及替加环素）的浓度。方法：50 μL血浆样本经甲醇沉淀蛋白，分别以头孢唑林、舒巴坦、多黏菌素E2及替加环素-d9为内标。色谱柱为Kinetex C18（3×100 mm，2.6 μm），流动相A为0.1%甲酸-水（含5 mmol·L-1乙酸铵），B为0.1%甲酸-甲醇，梯度洗脱，流速为0.5 mL·min-1，分析时间为5 min。采用电喷雾离子源，正负离子切换多反应监测模式扫描，其中头孢他啶：m/z 547.1→468.0（+），阿维巴坦：m/z 263.9→80.0（-），多黏菌素B1：m/z 602.6→101.2（+），多黏菌素B2：m/z 595.5→202.1（+），替加环素：m/z 586.4→513.4（+）。结果：血浆样本中各药物在一定范围内线性良好（r＞0.995 0），准确度为88.32%~110.42%，精密度RSD＜15%（定量下限处＜20%）。基质效应、提取回收率、稳定性、稀释可靠性和残留效应考察符合要求。运用本方法对27例碳青霉烯耐药革兰氏阴性菌感染患者共计64例样本进行血药浓度测定。结论：本方法操作简单快捷，准确灵敏，稳定性良好，适用于重症患者血浆中头孢他啶阿维巴坦、多黏菌素B及替加环素的治疗药物监测，可为临床碳青霉烯耐药革兰氏阴性菌感染治疗中的个体化给药提供科学参考。

Abstract:

Objective: To establish an LC-MS/MS method for simultaneous determination of the concentration of ceftazidime-avibactam, polymyxin B and tigecycline (containing ceftazidime, avibactam, polymyxin B1, B2 and tigecycline) in human plasma. Methods: The plasma samples (50 μL) were precipitated with methanol. Cefazolin, sulbactam, polymyxin E2 and tigecycline-d9 were used as internal standards. The chromatographic separation was performed on a Kinetex C18 column (3 mm×100 mm, 2.6 μm) with gradient elution using a mobile phase of 0.1% formic acid-water (containing 5 mmol·L-1 ammonium acetate solution) and 0.1% formic acid-methanol, at a flow rate of 0.5 mL·min-1. The total analysis time was 5 min. The detection of the analytes was performed by electrospray ionization, and the positive and negative ions were simultaneously switched and scanned. Multiple reaction monitoring was used with the transition of m/z 547.1→468.0 (ceftazidime, +), m/z 263.9→80.0 (avibactam, -), m/z 602.6→101.2 (polymyxin B1, +), m/z 595.5→202.1 (polymyxin B2, +), m/z 586.4→513.4 (tigecycline, +). Results: It was linear (r > 0.995 0) over the calibration range for different analytes. The accuracy was 88.32%~110.42%. The RSD of precision were less than 15% (20% at the lower limit of quantification). The matrix effect, recovery, stability, dilution integrity and carryover met the acceptance criteria. The plasma concentrations of three antibiotics in 64 samples from 27 patients with carbapenem-resistant organism infection were determined by this method. Conclusion: This method is simple and fast with good accuracy, sensitivity and stability. It is suitable for clinical therapeutic drug monitoring, providing scientific reference for individualized medication in the treatment of carbapenem-resistant organism infection.