食品中贝类过敏原的质谱定量分析方法研究  

Development of Mass Spectrometry Quantitative Analysis Method for Shellfish Allergens in Food

作　　者：杨沛洁 孟俊宏 赵晗 林浩鹏 关文强[1] 王凤玲[1] 杨术鹏 李熠 YANG Peijie;MENG Junhong;ZHAO Han;LIN Haopeng;GUAN Wenqiang;WANG Fengling;YANG Shupeng;LI Yi(Tianjin Key Laboratory of Food Biotechnology,School of Biotechnology and Food Science,Tianjin University of Commerce,Tianjin 300134;Institute of Food Science and Technology,Chinese Academy of Agricultural Sciences,Beijing 100193)

机构地区：[1]天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津300134 [2]中国农业科学院农产品加工研究所,北京100193

出　　处：《核农学报》2025年第2期329-338,共10页Journal of Nuclear Agricultural Sciences

基　　金：中国农业科学院青年创新专项(Y2023QC13);中国农业科学院农产品加工研究所青年创新专项(CAAS-ASTIP-Q2022-IFST-01)。

摘　　要：鉴于贝类是重要的过敏食物,为健全食品中贝类过敏原的定量分析方法、精准识别与检测食品中的贝类过敏原,本研究基于靶向蛋白质谱定量分析技术,开发了预制食品中蛤蜊过敏原的定量分析方法:样品经蛋白质的提取和酶解后,通过液相串联高分辨质谱和Peaks软件进行数据采集与分析;针对蛤蜊中的原肌球蛋白,经系统考察其酶解后肽段的特异性、质谱信号丰度、热稳定性等,筛选出ITELSEELAVVGNNCK作为最佳定量肽段;针对筛选出的肽段,采用液相串联三重四极杆质谱构建蛤蜊过敏原的定量分析方法,并进行系统的方法验证。结果表明,该方法具有良好的特异性、灵敏度[定量限(LOQ)为10 mg·kg^(-1)]、准确度(回收率为78%~105.3%)和精密度(日内和日间变异系数均<13.0%)。用该法检测分析实际样品发现,部分预制食品中存在蛤蜊过敏原的污染,但未进行明确标识。本研究开发的方法可为食品中贝类过敏原的风险监测提供技术和方法支撑。Shellfish is an important allergenic food.To improve the quantitative analysis methods for shellfish allergens in food,and accurately identify and detect shellfish allergens in food,a quantitative analysis method for clam allergens in prepared foods was developed based on targeted proteomic quantitative analysis technology.After protein extraction and enzymatic digestion of samples,liquid chromatography-tandem high-resolution mass spectrometry and Peaks software were used for data acquisition and analysis.For tropomyosin in clams,ITELSEELAVVGNNCK was screened as the best peptide for quantification by systematically examining the specificity of its enzymatic peptide,mass spectrometry signal abundance,and thermal stability.Liquid tandem triple quadrupole mass spectrometry was used to construct a quantitative method for the analysis of clam allergens against the screened peptides.A systematic method validation was carried out,which demonstrated that the developed method possessed good specificity,sensitivity[Limit of Quantification(LOQ)of 10 mg·kg^(-1)],accuracy(recoveries of 78%-105.3%),and precision(the intra-day and inter-day coefficients of variation<13.0%).Testing and analyzing actual samples by this method revealed that contamination with clam allergens was present in some prepared foods without clear labelling.The method developed in this study could provide technical and methodological support for risk monitoring of shellfish allergens in food.

关 键 词：过敏原 贝类 蛤蜊 定量分析 质谱分析 

分 类 号：O657.63[理学—分析化学] TS254.7[理学—化学]