超高效液相色谱-飞行时间质谱法鉴定鹅源性成分特征肽及其应用研究  

Application and Identification of Goose-Derived Specific Peptides by Ultra-high Performance Liquid Chromatography-Time of Flight Mass Spectrometry

作　　者：李碧芳[1] 王韦达[1] 刘奕雄[1] 李意[1] 程诚 陈序 杜业刚[1] LI Bi-fang;WANG Wei-da;LIU Yi-xiong;LI Yi;CHENG Cheng;CHEN Xu;DU Ye-gang(Shenzhen Academy of Metrology and Quality Inspection,National Nutrition Food Testing Center(Guangdong),Shenzhen 518109;College of Public Health,Zhengzhou University,Zhengzhou 450001)

机构地区：[1]深圳市计量质量检测研究院,国家营养食品质量监督检验中心(广东),广东深圳518109 [2]郑州大学公共卫生学院,河南郑州450001

出　　处：《分析科学学报》2019年第5期629-634,共6页Journal of Analytical Science

基　　金：深圳市科技计划(No.YCYJ20170817162416978);广东省质量技术监督局科技项目(No.2016PZ12);深圳市计量质量检测研究院科技项目(No.2018-YA49)

摘　　要：本文建立了一种鉴定鹅源性成分的高效液相色谱-串联质谱分析方法。样品经过蛋白质提取、蛋白质的还原和烷基化、胰蛋白酶水解后,利用纳升液相色谱-飞行时间质谱和ProteinPilotTM软件分别进行多肽的检测和鉴定。通过BLAST搜索与Uniprot数据库对比分析,筛选出鹅肉的备选物种特征肽,再用超高效液相色谱-串联质谱的多反应监测(MRM)模式进行定量和验证。各特征肽段定量限为3.5~6.0 fmol,鸭肉中鹅肉成分的线性相关系数大于0.99,定量限为5%。A method based on ultra-high performance liquid chromatography-tandem mass spectrometry has been established for identification of goose-derived components.The sample went through protein extraction,protein reduction and alkylation,and tryptic digestion in turn.Peptide determination was conducted by the aid of a nanoLC-Triple-TOF MS system and peptide identification was performed using ProteinPilotTM software.Potential species marker peptides for goose were screened with the blast tool against the Uniprot database.The specificity of peptides was further confirmed by UPLC-MS/MS with multiple-reaction monitoring(MRM) mode,and then quantification of goose in duck was also studied.The limit of quantitation for goose was 5% and the linear correlation coefficient was more than 0.99.Moreover,absolute limits of quantitation for marker peptides were 3.5-6.0 fmol.This reported method could be used to authenticate goose exclusively.

关 键 词：蛋白质组学 特征肽段 鹅肉 质谱 

分 类 号：O657.63[理学—分析化学]