色谱

色谱 ›› 2017, Vol. 35 ›› Issue (10): 1037-1041.DOI: 10.3724/SP.J.1123.2017.06013

• 研究论文 • 上一篇    下一篇

基于高分辨质谱技术的婴幼儿食品中过敏原蛋白质的高灵敏检测

吴慈1, 陈溪1, 刘健慧2,3, 张晓林1, 薛伟锋1, 梁振2, 刘梦遥1, 崔妍1, 黄大亮1, 张丽华2   

  1. 1. 大连出入境检验检疫局检验检疫技术中心, 辽宁 大连 116100;
    2. 中国科学院分离分析化学重点实验室, 国家色谱研究分析中心, 中国科学院大连化学物理研究所, 辽宁 大连 116023;
    3. 中国科学院大学, 北京 100049
  • 收稿日期:2017-06-07 出版日期:2017-10-08 发布日期:2017-10-18
  • 通讯作者: 张丽华,E-mail:lihuazhang@dicp.ac.cn.
  • 基金资助:

    中国科学院分离分析化学重点实验室开放基金项目（KL1606）；国家质量监督检验检疫总局科技计划项目（2015IK343）

High-sensitive detection of multiple allergenic proteins in infant food with high-resolution mass spectrometry

WU Ci1, CHEN Xi1, LIU Jianhui2,3, ZHANG Xiaolin1, XUE Weifeng1, LIANG Zhen2, LIU Mengyao1, CUI Yan1, HUANG Daliang1, ZHANG Lihua2   

  1. 1. Inspection and Quarantine Technical Center of Dalian Entry-Exit Inspection and Quarantine Bureau, Dalian 116100, China;
    2. Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;
    3. University of the Chinese Academy of Sciences, Beijing 100049, China
  • Received:2017-06-07 Online:2017-10-08 Published:2017-10-18
  • Supported by:

    Open Fund Supported by Key Laboratory of Separation Sciences for Analytical Chemistry, Chinese Academy of Sciences (No. KL1606); Science and Technology Project of State General Administration of the People's Republic of China for Quality Supervision and Inspection and Quarantine (No. 2015IK343).

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摘要：

基于高效液相色谱-串联质谱（LC-MS/MS）技术，选择稳定性好、灵敏度高的特征肽段，利用平行反应监测（PRM）技术，实现了多类过敏原蛋白质的高灵敏度同时检测，并成功应用于婴幼儿食品中过敏原成分的分析。对于婴幼儿食品中蛋白质的提取，与传统的丙酮沉淀法比，采用膜上原位样品预处理方法（i-FASP）可实现更高的蛋白质提取效率和抗干扰能力。所检测的过敏原蛋白质的定量限（LOQ）最小可达到0.028 mg/L，其线性范围最宽可跨越4个数量级，且线性关系良好（相关系数R2≥0.99）。该方法为食品中过敏原蛋白质组学快速分析提供了一种可靠的分析方法。

关键词: 过敏原蛋白质, 平行反应监测, 液相色谱-串联质谱, 婴幼儿食品

Abstract:

A novel method of the simultaneous detection of multiple kinds of allergenic proteins in infant food with parallel reaction monitoring (PRM) mode using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was established. In this method, unique peptides with good stability and high sensibility were used to quantify the corresponding allergenic proteins. Furthermore, multiple kinds of allergenic proteins are inspected simultaneously with high sensitivity. In addition, such method was successfully used for the detection of multiple allergenic proteins in infant food. As for the sample preparation for infant food, compared with the traditional acetone precipitation strategy, the protein extraction efficiency and capacity of resisting disturbance are both higher with in-situ filter-aided sample pretreatment (i-FASP) method. All allergenic proteins gave a good linear response with the correlation coefficients (R2) ≥ 0.99, and the largest concentration range of the allergenic proteins could be four orders of magnitude, and the lowest detection limit was 0.028 mg/L, which was better than that reported in references. Finally, the method was conveniently used to detect the allergens from four imported infant food real samples. All the results demonstrate that this novel strategy is of great significance for providing a rapid and reliable analytical technique for allergen proteomics.

Key words: allergenic protein, infant food, liquid chromatography-tandem mass spectrometry (LC-MS/MS), parallel reaction monitoring (PRM)

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