色谱

色谱 ›› 2018, Vol. 36 ›› Issue (12): 1269-1278.DOI: 10.3724/SP.J.1123.2018.08005

• 研究论文 • 上一篇    下一篇

基于液相色谱-串联质谱法的肉类特征肽段鉴别及掺假测定

古淑青1, 詹丽娜2, 赵超敏1, 郑江1, 蔡一村1, 邓晓军1   

  1. 1. 上海出入境检验检疫局动植物与食品检验检疫技术中心, 上海 200135;
    2. 上海大学生命科学学院食品工程系, 上海 200436
  • 收稿日期:2018-08-03 出版日期:2018-12-08 发布日期:2014-11-06
  • 通讯作者: 邓晓军,Tel:(021)38620526,E-mail:dengxj@shciq.gov.cn
  • 基金资助:

    上海市中央引导地方科技发展专项项目（YDZX20173100004528）；上海市技术标准专项项目（17DZ2201100）；长三角科技合作项目（17395810102）.

Identification of meat marker peptides and detection of adulteration by liquid chromatography-tandem mass spectrometry

GU Shuqing1, ZHAN Lina2, ZHAO Chaomin1, ZHENG Jiang1, CAI Yicun1, DENG Xiaojun1   

  1. 1. Technical Center for Animal Plant and Food Inspection and Quarantine, Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China;
    2. Department of Food Engineering, School of Life Science, Shanghai University, Shanghai 200436, China
  • Received:2018-08-03 Online:2018-12-08 Published:2014-11-06
  • Supported by:

    Shanghai Central Guiding Local Science and Technology Development Special Project (No. YDZX20173100004528); Shanghai Technical Standard Project (No. 17DZ2201100); Science and Technology Joint Project of the Yangtze River Delta (No. 17395810102).

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摘要：

建立了液相色谱-串联质谱技术鉴别肉类特征肽段及定量检测羊肉中常见外源肉掺假的方法。样品经蛋白质提取、胰蛋白酶水解和固相萃取小柱净化后，利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱（UPLC-Q/Exactive-HRMS）和Proteinpilot软件，实现蛋白质和多肽的鉴定；再通过基本局部比对搜索工具（BLAST）与Uniprot数据库对比分析，筛选出羊肉、鸭肉、猪肉和鸡肉的20个物种特征性多肽标志物；最后利用高效液相色谱-三重四极杆质谱（UPLC-QqQ-MS）系统对羊肉、鸭肉、猪肉和鸡肉的特征性多肽进行了验证和多反应监测（MRM）定量研究。将鸭肉、猪肉和鸡肉分别按照质量分数为1%、5%、10%、20%、50%的比例掺加到羊肉中，得到鸭肉最低掺假检出限为0.25%、猪肉最低掺假检出限为0.17%、鸡肉最低掺假检出限为0.10%。

关键词: 掺假测定, 鉴别, 肉, 特征肽段, 液相色谱-串联质谱

Abstract:

A liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the identification of meat marker peptides and quantitative detection of common exogenous meat in mutton adulteration. Samples were prepared by protein extraction, trypsin hydrolysis, and solid phase extraction. Proteins and peptides were identified using ultra-performance liquid chromatography-quadrupole/electrostatic orbitrap-high resolution mass spectrometry (UPLC-Q/Exactive-HRMS) combined with Proteinpilot software. Twenty species-specificity marker peptides in mutton, duck, pork, and chicken were identified by comparison of the basic local alignment search tool (BLAST) with the Uniprot database. Verification and multiple reaction monitoring (MRM) of these marker peptides were performed quantitatively using a UPLC-triple-quadrupole mass spectrometry (QqQ-MS) system. Duck, pork, and chicken were added to mutton at mass percentages of 1%, 5%, 10%, 20%, and 50%. The limit of detection for the adulterants was 0.25% for duck, 0.17% for pork, and 0.10% for chicken.

Key words: adulteration detection, identification, liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), marker peptides, meat

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