基于液质联用技术的脂肪酸修饰型GLP-1类似物一级结构测定  

Measurement of primary structure of fatty acid modified GLP-1 analogues by LC-MS/MS

作　　者：胡馨月 孙悦 李懿 吕萍 张慧 李晶 梁成罡 HU Xin-yue;SUN Yue;LI Yi;LYU Ping;ZHANG Hui;LI Jing;LIANG Cheng-gang(Division of Hormone,Institute of Chemical Drug Control,National Institutes for Food and Drug Control,NHC Key Laboratory of Research on Quality and Standardization of Biotech Products,NMPA Key Laboratory for Quality Research and Evaluation of Chemical Drugs,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院化学药品检定所激素室/国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室/国家药品监督管理局化学药品质量研究与评价重点实验室,北京102629

出　　处：《中国医药生物技术》2023年第6期540-549,共10页Chinese Medicinal Biotechnology

基　　金：中国食品药品检定研究院中青年发展研究基金(2022A3)。

摘　　要：目的使用超高效液相色谱-四极杆串联飞行时间质谱法(UPLC-Q-TOF MS)对脂肪酸修饰型胰高血糖素-1(GLP-1)类似物利拉鲁肽和司美格鲁肽一级结构进行测定。方法分别在完整分子和酶切肽段水平上检测,采用ACQUITY UPLC peptide BEH C18(2.1 mm×100 mm,30 nm,1.7μm)色谱柱,流动相A为0.1%甲酸-水溶液,流动相B为0.1%甲酸-乙腈溶液,梯度洗脱,流速0.3 ml/min;采用正离子ESI+离子源和MSE采集模式进行数据采集。结果分别从完整分子和肽段水平准确测定了两种脂肪酸修饰型GLP-1类似物一级完整序列结构信息,并对修饰侧链位点进行准确定位。肽段覆盖率为100%,且专属性强、重复性好。结论液质联用技术快速、灵敏、准确,可用于GLP-1类似物一级结构的表征分析。Objective The primary structures of fatty acid-modified GLP-1 analogues liraglutide and semaglutide were determined by UPLC-Q-TOF MS.Method The levels of intact molecule and enzyme-ligated peptide were detected.ACQUITY UPLC peptide BEH C18(2.1 mm×100 mm,30 nm,1.7μm);mobile phase A was formic acid-water(1:1000,V/V),mobile phase B was formic acid-acetonitrile(1:1000,V/V),gradient elution,flow rate 0.3 ml/min;MSE mode monitor was carried out by ESI+scanning.Results The first order complete sequence information structure of two fatty acid modified GLP-1 analogues was accurately determined from the level of intact molecule and peptide,and the modified side chain sites were accurately located.The peptide coverage was 100%,and the method was specific and reproducible.Conclusion LC-MS/MS technology is a rapid,sensitive and accurate method to analyze the primary structure of GLP-1 analogues.

关 键 词：GLP-1类似物 利拉鲁肽 司美格鲁肽 液质联用技术 一级结构 N末端序列 

分 类 号：TQ460.1[化学工程—制药化工] O657.63[理学—分析化学]