超高效液相色谱串联质谱法测定人血清中妥布霉素浓度及其吸入剂在支气管扩张患者中的药代动力学研究  

Establishment of ultra-performance liquid chromatography-tandem mass spectrometry method for assay of tobramycin in human serum and its use in a pharmacokinetic study of tobramycin inhalation in bronchiectasis patients

作　　者：王雨 徐晓勇 黄晓岚 刘笑芬 范亚新 胡佳丽 毋海兰 张菁 郭蓓宁 WANG Yu;XU Xiaoyong;HUANG Xiaolan;Liu Xiaofen;FAN Yaxin;HU Jiali;WU Hailan;ZHANG Jing;Guo Beining(Institute of Antibiotics,Huashan Hospital,Fudan University,Key Laboratory of Clinical Pharmacology of Antibiotics,Ministry of Health,National Clinical Research Center for Aging and Medicine,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院抗生素研究所,国家卫生健康委员会抗生素临床药理重点实验室,国家老年疾病临床医学研究中心,上海200040

出　　处：《中国感染与化疗杂志》2024年第5期545-552,共8页Chinese Journal of Infection and Chemotherapy

基　　金：国家重点研发计划资助(2020YFC2005000);上海市领军人才(LJ2016-01);十三五国家科技重大专项(2017ZX09201002-002)。

摘　　要：目的建立准确测定人血清中妥布霉素浓度的超高效液相色谱串联质谱法(UPLC-MS/MS),并将该法应用于妥布霉素吸入剂的临床药代动力学研究。方法以妥布霉素-D12为内标,采用固相萃取法预处理血清样本。色谱柱为TitankHilic 3μm(2.1 mm×100 mm,3µm)柱。流动相由0.1%甲酸-乙腈(A相)和0.1%甲酸水溶液(B相)组成,流速为0.4 mL/min,梯度洗脱。采用电喷雾电离源,正离子方多反应监测(MRM)扫描进行监测,定量离子对分别为m/z 468.3→m/z 163.3(妥布霉素)和m/z 480.6→m/z 166.2(妥布霉素-D12)。考察该方法的选择性、交互影响、合并用药、标准曲线和定量下限、精密度和准确度、回收率、基质效应及稳定性等。结果妥布霉素在0.0500~10.0 mg/L内线性良好(R^(2)=0.9995);批内、批间精密度良好(变异系数均≤3.6%),准确度偏差在-0.4%~6.0%范围内。人血清(包含溶血和脂血)中妥布霉素内标校正的基质效应因子(MF)为92.2%~94.9%,变异系数均≤2.7%;妥布霉素血清样品提取回收率为79.5%~81.9%,内标提取回收率为78.9%;血清样品在室温、-20℃/-70℃下分别放置72 h、274 d稳定。妥布霉素吸入剂在患者中药代动力学研究结果显示,给药剂量为300 mg/次,每日2次,第1天和第28天连续给药后测得3例患者的C_(max)分别为(0.72±0.61)mg/L和(0.76±0.73)mg/L,T_(max)分别为(1.83±0.61)h和(1.50±0.50)h。结论本方法灵敏度高、准确性好且快速,适用于妥布霉素吸入剂临床药代动力学研究和妥布霉素的治疗药物浓度监测。Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of tobramycin in human serum,and examine the utility of the method in a clinical pharmacokinetic study of tobramycin inhalation.Methods Serum samples were pretreated by solid phase extraction with tobramycin-D12 as internal standard.Chromatographic separation was performed on a TitankHilic(2.1 mm×100 mm,3µm)column.The mobile phase consisted of 0.1%formic acid-acetonitrile and 0.1%formic acid aqueous solution at a flow rate of 0.4 mL/min.Electrospray ionization source and multiple reaction monitoring(MRM)scanning were used for monitoring the quantitative ion pairs with m/z 468.3→m/z 163.3(tobramycin)and m/z 480.6→m/z 166.2(tobramycin-D12).The established method was investigated in terms of selectivity,interaction,concomitant medication,standard curve and lower limit of quantitation,precision and accuracy,recovery,matrix effect,and stability of tobramycinin.Results The linear range of tobramycin was 0.0500-10.0 mg/L(R^(2)=0.9995).The intra-and inter-batch precision was satisfactory(coefficient of variation[CV]≤3.6%).The accuracy ranged from-0.4%to 6.0%.The matrix effect factor(MF)in human serum samples(including hemolysis and lipemia)ranged from 92.2%to 94.9%(CV≤2.7%).The recovery of tobramycinin was 79.5%-81.9%in serum samples,while the recovery of internal standard was 78.9%.The analyte was stable in serum samples for 72 h at room temperature and for 274 days at-20℃/-70℃.The pharmacokinetic study of tobramycin inhalation in bronchiectasis patients showed that after continuous administration of tobramycin 300 mg twice a day to 3 patients,the mean C_(max) of tobramycin was(0.72±0.61)mg/L on Day 1 and(0.76±0.73)mg/L on Day 28,respectively.The corresponding T_(max) was(1.83±0.61)h and(1.50±0.50)h,respectively.Conclusions The UPLC-MS/MS method established in this study is sensitive,accurate and rapid.It is successfully applied to the clinical pharmacokinetic study of tobramyc

关 键 词：妥布霉素 超高效液相色谱串联质谱法 血药浓度 

分 类 号：R978.12[医药卫生—药品]