Meng Jia, Gu Shuqing, Fang Zhen, Niu Bing, Deng Xiaojun, Guo Dehua, Zhu Jian, Han Fang
Department of Food Engineering, School of Life Science, Shanghai University, Shanghai 200444, China.
Technical Center for Animal Plant and Food Inspection and Quarantine, Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China.
Se Pu. 2019 Jul 8;37(7):712-722. doi: 10.3724/SP.J.1123.2019.01042.
A liquid chromatography-tandem mass spectrometry method for the identification of marker peptides of aquatic product allergens and quantitative detection of multiple allergens in meat products and seasonings was developed. The samples were prepared by protein extraction, protein purification, and trypsin hydrolysis. The proteins and peptides were identified using ProteinPilot by the data analysis of the ion spectrum of polypeptide fragments using ultra-performance liquid chromatography-quadrupole/electrostatic orbitrap high-resolution mass spectrometry (UPLC-Q/Exactive-HRMS). The identification of 30 species-specific marker peptides in , and by comparison of the basic local alignment search tool (BLAST) with the UniProt database was achieved. The verification and multiple reaction monitoring (MRM) quantitative studies of these marker peptides were performed using a triple quadrupole mass spectrometry (UPLC-QqQ-MS) system. The proposed method showed a good linear relationship in the range of 5-250 mg/kg. The limits of quantitation and observed recoveries were in the range of 2-3.5 mg/kg and 88.7%-110.2%, respectively. This method presents various advantages such as good repeatability and high throughput, suitability for rapid screening, and quantitative analysis of seven aquatic allergens in meat products and seasonings.
建立了一种液相色谱-串联质谱法,用于鉴定水产品过敏原的标记肽,并定量检测肉制品和调味料中的多种过敏原。通过蛋白质提取、蛋白质纯化和胰蛋白酶水解制备样品。使用ProteinPilot通过超高效液相色谱-四极杆/静电轨道阱高分辨率质谱(UPLC-Q/Exactive-HRMS)对多肽片段的离子谱进行数据分析来鉴定蛋白质和肽。通过将基本局部比对搜索工具(BLAST)与UniProt数据库进行比较,实现了对30种特异性标记肽的鉴定。使用三重四极杆质谱(UPLC-QqQ-MS)系统对这些标记肽进行验证和多反应监测(MRM)定量研究。所提出的方法在5-250 mg/kg范围内显示出良好的线性关系。定量限和观察到的回收率分别在2-3.5 mg/kg和88.7%-110.2%范围内。该方法具有重复性好、高通量、适用于快速筛选以及对肉制品和调味料中的七种水产过敏原进行定量分析等多种优点。