分析测试学报2025，Vol.44Issue(10)：2162-2168,7.DOI:10.12452/j.fxcsxb.24120905
胰酶酶解/高效液相色谱-四极杆-飞行时间质谱法检测虾制品中过敏原蛋白
Detection of Allergenic Protein from Shrimp Products by Pancreatic Enzymolysis/High Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry
摘要
Abstract
In this study,an analytical method was established for determination of allergenic protein in shrimp products based on characteristic peptides by pancreatic enzymolysis/high performance liq-uid chromatography-quadrupole time of flight mass spectrometry.With 100 μL 6 mol/L guanidine hy-drochloride and 700 μL 50 mmol/L Tris-HCl buffer as denaturating and extraction reagents,50 mmol/L dithiositol and 100 mmol/L iodoacetamide were used to destroy and inhibit the formation of di-sulfide bonds by optimizing the enzymolysis time and the selection of characteristic peptide.After en-zymolysis with 10 mg/mL trypsin in a weak alkaline environment created by 100 μL and 500 mol/L ammonium bicarbonate,the analytes were separated by using a Caprisil C18-X(100 mm×2.1 mm,1.8 μm)column with 0.1%formic acid aqueous solution and acetonitrile as the mobile phase for elu-tion.The ionization was performed by Dual AJS ESI in positive mode,then the ions were detected by quadrupole time of flight mass spectrometry,with cyclic sampling performed by the Auto MS/MS ion acquisition iterative exclusion mode.IQLLEEDLER was selected as the characteristic peptide of tropomyosin according to chromatographic results.The results showed that the characteristic peptides selected in the experiment had good linear relationship in the linear range,the correlation coefficient(r2)was 0.992 0,the limit of detection was 0.68 μg/L,and the limit of quantitation was 2.28 μg/L.The recoveries of the samples were 84.5%-101%at 2 and 10 mg/kg standard levels.The method is simple,sensitive,reproducible and accurate,which can provides technical support for the identifi-cation of shrimp allergens and method reference for the identification of animal-derived components based on characteristic peptides.关键词
胰酶酶解/高效液相色谱-四极杆-飞行时间质谱法/虾制品/过敏原蛋白/特征肽段Key words
pancreatic enzymolysis/high performance liquid chromatography-quadrupole-time of flight mass spectrometry/shrimp products/allergenic protein/characteristic peptide segments分类
化学化工引用本文复制引用
史洪飞,郭明鑫,周修齐,徐伯芃,周用武,徐成鑫,周亦飞,冶金,张吕悦,刘柯宇..胰酶酶解/高效液相色谱-四极杆-飞行时间质谱法检测虾制品中过敏原蛋白[J].分析测试学报,2025,44(10):2162-2168,7.基金项目
国家重点研发计划项目(2023YFC3304000) （2023YFC3304000）
2025年省级大学生创新重点项目(X202512263052) （X202512263052）
