食品科学 ›› 2018, Vol. 39 ›› Issue (20): 332-336.doi: 10.7506/spkx1002-6630-201820048

• 安全检测 • 上一篇    下一篇

超高效液相色谱-串联质谱法检测食品中鸡蛋过敏原卵白蛋白

宁亚维1,刘茁1,范素芳2,马俊美2,李强2,*,张岩2,*   

  1. (1.河北科技大学生物科学与工程学院,河北?石家庄 050018;2.河北省食品检验研究院,河北?石家庄 050000)
  • 出版日期:2018-10-25 发布日期:2018-10-24
  • 基金资助:
    国家重大科学仪器设备开发专项(2012YQ03011110);河北省自然科学基金资助项目 (C2015321005)

Detection of Egg Allergen Ovalbumin in Food by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

NING Yawei1, LIU Zhuo1, FAN Sufang2, MA Junmei2, LI Qiang2,*, ZHANG Yan2,*   

  1. (1. College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang 050018, China; 2. Hebei Food Inspection and Research Institute, Shijiazhuang 050000, China)
  • Online:2018-10-25 Published:2018-10-24

摘要: 建立超高效液相色谱-串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)检测食品中鸡蛋过敏原卵白蛋白的方法。蛋清经胰蛋白酶酶解、超滤净化,经Easy-nLC 1000纳升液相色谱-四极杆-静电场轨道阱高分辨质谱以乙腈-0.1%甲酸溶液作为流动相进行分离,结合Uniprot数据库筛选出特异性好且灵敏度高的特征肽段GGLEPINFQTAADQAR(GR-16)作为外标,采用UPLC-MS/MS正离子源多反应监测模式对实际样品进行检测。结果表明,该方法在5~5?000?ng/mL范围内线性良好,定量限为0.1?μg/g,平均回收率在77.22%~98.55%之间,相对标准偏差不大于9.97%。此方法特异性高、灵敏度好、分析速度快,可为食品中鸡蛋过敏原的检测提供技术支持。

关键词: 超高效液相色谱-串联质谱, 过敏原, 卵白蛋白, 胰蛋白酶, 特征肽段?

Abstract: An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the detection of the egg allergen ovalbumin in food in this investigation. Egg white was trypsinized and the resulting hydrolysate was purified by ultrafiltration, and then separated by Easy-nLC 1000 nano liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry with acetonitrile-0.1% (V/V) formic acid water as the mobile phase. By comparison with the Uniprot database, the characteristic peptide GGLEPINFQTAADQAR(GR-16) with good specificity and sensitivity was selected. Then, the UPLC-MS/MS analysis was performed using positive ion electrospray ionization (ESI+) in the multi-reaction monitoring (MRM) mode. The method showed a good linear relationship in the concentration range of 5–5 000 ng/mL with a limit of quantification (LOQ) of 0.1 μg/g. The average recoveries ranged from 77.22% to 98.55% with relative standard deviations (RSDs) lower than 9.97%. This method has high specificity and good sensitivity and is also time-saving, and thus it can provide technical support for the accurate determination of egg allergens in food product.

Key words: ultra-high performance liquid chromatography-tandem mass spectrometry, allergen, ovalbumin, trypsin, characteristic peptides

中图分类号: