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(5μm,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1~100μg·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3.61~19.60μg·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。
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 denaturation,reduction and alkylation,trypsin was added to digest infliximab into a large number of peptides.Quantification was performed by analyzing signature peptide,and peptide of bevacizumab was used as the internal standard.The chromatographic column was Ultimate LP C18(5μm,21 mm×100 mm),and the mobile phases were water containing 0.1%formic acid and acetonitrile 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 1-100μg·mL^(-1).The intra-and inter-batch accuracy ranged from 85.0%-115.0%,and the intra-and inter-batch precision was within 15.0%.Specificity,matrix effect,stability,selectivity and carryover all met the requirements.The plasma concentrations of infliximab in 10 patients with inflammatory bowel disease ranged from 3.61 to 19.60μg·mL^(-1).CONCLUSION The LC-MS/MS method established in our study can accurately and precisely determine the plasma concentration of infliximab,which provides an effective means for the clinical monitoring of infliximab.
作者
李宁红
向东
刘璐
余恒毅
李喜平
邱丽慧
赵婷慧
贡雪芃
LI Ninghong;XIANG Dong;LIU Lu;YU Hengyi;LI Xiping;QIU Lihui;ZHAO Tinghui;GONG Xuepeng(Department of Pharmacy,Affiliated Tongji Hospital,Tongji Medical Collage,Huazhong University of Science and Technology,Hubei Wuhan 430030,China)
出处
《中国医院药学杂志》
CAS
北大核心
2024年第5期506-512,共7页
Chinese Journal of Hospital Pharmacy
基金
湖北省自然科学基金项目(编号:2022CFB142)。
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
infliximab
therapeutic drug monitoring
liquid chromatography tandem-mass spectrometry
blood drug concentration detection
