液相色谱串联质谱法测定人血浆中英夫利昔单抗浓度
Determination of infliximab in human plasma by liquid chromatography tandem-mass spectrometry
李宁红 1向东 1刘璐 1余恒毅 1李喜平 1邱丽慧 1赵婷慧 1贡雪芃1
作者信息
- 1. 华中科技大学同济医学院附属同济医院药学部,湖北武汉 430030
- 折叠
摘要
目的:建立液相色谱串联质谱法(liquid chromatography tandem-mass spectrometry,LC-MS/MS)测定人血浆中英夫利昔单抗血药浓度的方法.方法:采用甲醇沉淀提取血浆中的英夫利昔单抗,经变性、还原、烷基化后,加入胰酶裂解成大量肽段,通过分析特征性肽段并进行测定,选用贝伐珠单抗特征肽段作为内标.色谱柱为Ultimate LP C18(5 μm,21 mm× 100 mm),以含0.1％甲酸的水和含0.1％甲酸的乙腈为流动相进行梯度洗脱.采用电喷雾离子源,以多反应监测模式进行正离子扫描.对所建立的LC-MS/MS方法进行方法学验证并利用该方法测定患者的血浆药物浓度.结果:英夫利昔单抗在1～100 μg·mL-1范围内线性关系良好,批内和批间准确度在85.0％～115.0％,批内和批间精密度均小于15％,特异性、基质效应、稳定性、选择性、残留均符合指导原则要求.10例炎症性肠病患者体内英夫利昔单抗血药浓度在3.61～19.60 μg·mL-1范围内.结论:该研究建立的LC-MS/MS方法可以准确、精密测定英夫利昔单抗血药浓度,为临床监测英夫利昔单抗提供了有效的检测手段.
Abstract
OBJECTIVE To establish a liquid chromatography tandem mass spectrometry(LC-MS/MS)method for the quantification of infliximab in human plasma.METHODS Infliximab was extracted from plasma by methanol.After denatur-ation,reduction and alkylation,trypsin was added to digest infliximab into a large number of peptides.Quantification was per-formed by analyzing signature peptide,and peptide of bevacizumab was used as the internal standard.The chromatographic col-umn was Ultimate LP C18(5 μm,21 mm×100 mm),and the mobile phases were water containing 0.1％ formic acid and acetoni-trile containing 0.1％formic acid.The multiple reaction monitoring was performed in the positive electrospray ionization mode.The established LC-MS/MS method was validated and used to determine plasma drug concentrations of infliximab-treated patients.RESULTS Infliximab showed good correlation within the range of 1-100 μg·mL-1.The intra-and inter-batch accu-racy ranged from 85.0％-115.0％,and the intra-and inter-batch precision was within 15.0％.Specificity,matrix effect,stabil-ity,selectivity and carryover all met the requirements.The plasma concentrations of infliximab in 10 patients with inflammatory bowel disease ranged from 3.61 to 19.60 μg·mL-1.CONCLUSION The LC-MS/MS method established in our study can accu-rately and precisely determine the plasma concentration of infliximab,which provides an effective means for the clinical monitoring of infliximab.
关键词
英夫利昔单抗/治疗药物监测/液相色谱串联质谱/血药浓度检测Key words
infliximab/therapeutic drug monitoring/liquid chromatography tandem-mass spectrometry/blood drug concen-tration detection引用本文复制引用
基金项目
湖北省自然科学基金项目(2022CFB142)
出版年
2024
NETL
NSTL
万方数据