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液相色谱串联质谱法测定牛排产品中大豆蛋白含量

李莹莹,张颖颖,任南,李石磊,赵文涛,田寒友,王守伟   

  1. 中国肉类食品综合研究中心
  • 收稿日期:2019-10-14 修回日期:2020-03-20 出版日期:2020-05-25 发布日期:2020-05-15
  • 通讯作者: 李莹莹 E-mail:ying_sky@126.com
  • 基金资助:
    重要食品真实性多维鉴别检测关键技术研究

Determination of Soybean Protein in Steak Products by Liquid Chromatography-Tandem Mass Spectrometry

Ying-Ying LIZHANG Yingying 2,Shi-Lei Li 2,   

  • Received:2019-10-14 Revised:2020-03-20 Online:2020-05-25 Published:2020-05-15
  • Contact: Ying-Ying LI E-mail:ying_sky@126.com
  • Supported by:
    Research on key technologies of multi-dimensional identification detection of important food authenticity

摘要: 建立了液相色谱-串联质谱技术测定牛排产品中大豆蛋白含量的方法。首先利用高分辨质谱精确的质量数判别能力,结合Unitprot数据库,对大豆特异性多肽进行鉴别及筛选,获得可用于大豆准确定性的多肽序列信息,并利用skyline软件将多肽序列转换为离子对信息,再利用高效液相色谱-三重串联四极杆质谱(LC-MS/MS)对大豆蛋白特异性多肽进行确证,构建大豆蛋白LC-MS/MS定性判别方法,并进一步开展用于定量多肽的筛选研究,最终筛选到了线性较好,回收率满足的定量多肽共3条,分别是GSDLVNVR、VSDDEFNNYK、LVFCPQQAEDDK,线性相关系数均达到0.99以上,加标回收率为81.7%~115.8%,精密度小于13%,最低检出限为0.15g/100g,定量限为0.5g/100g,同时对市售28个牛排进行了大豆蛋白含量的测定,构建了液相色谱串联质谱方法测定牛排中大豆蛋白含量的方法。结果表明,该方法特异性好,准确度高,同时可推广应用至其他肉制品。

关键词: 液相色谱-串联质谱, 多肽, 牛排, 大豆蛋白, 定量

Abstract: A method for the determination of soybean protein in steak products by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. Firstly, the soybean specific peptides were identified and screened based on the high-resolution mass spectrometry with the accurate mass resolution and the Unitprot database. Then, the soybean peptide sequences were converted into ion information by the skyline software. Next, the soybean specific peptides were confirmed by LC-MS/MS, and the soybean peptides for quantification were further carried out. Finally, 3 quantitative peptides with good linearity and recovery rate were selected, which were GSDLVNVR, VSDDENNNYK, and LVCCPQQAEDDK. The linear correlation coefficients were all above 0.99, the recoveries were 81.7%~115.8%, and the precision was less than 13%. The minimum detection limit was 0.15%, the limit of quantification was 0.5%, and the soybean protein contents of 28 commercially available steaks were determined. The results showed that the method had good specificity and high accuracy, and can be applied to other meat products.

Key words: liquid chromatography tandem mass spectrometry (HPLC-MS/MS), peptide identification, steak, soybean protein, quantitative

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