Technical Center, Qingdao Customs, Qingdao, People's Republic of China.
College of Food Science and Engineering, Ocean University of China, Qingdao, People's Republic of China.
J Sci Food Agric. 2021 Sep;101(12):5278-5285. doi: 10.1002/jsfa.11177. Epub 2021 Mar 18.
Allergic reactions to crustacean products have been increasing owing to the rising consumption. Tropomyosin (TM) is the main crustacean allergen; it has a coiled-coil structure, which shows stability to various food processing methods. Crustacean processed products have been used in several food products, thereby causing greater difficulties in detecting TM in these products. We aimed to develop an assay based on high-performance liquid chromatography-tandem mass spectrometry for the accurate and reproducible quantification of crustacean TM in foods.
The three peptides IQLLEEDLER, LAEASQAADESER, and IVELEEELR were selected as peptide markers, and the peptide IVELEEELR was selected as the quantitative marker. Extraction conditions and enzymatic digestion conditions were completely optimized. The extraction solution of Tris-hydrochloric acid buffer (50 mmol L , pH 7.4) containing 1 mol L potassium chloride and the enzymatic treatment at 1:15 ratio (enzyme/protein, m/m) for 13 h showed excellent efficiency. The method exhibited a good linear relationship, with the qualified coefficient of determination (R = 0.9994) in the wide range of 1 to 1000 μg L . The accuracy was validated based on spiked recovery at three spiking levels (12.5, 25.0, and 50.0 μg kg , TM/matrix) in blank matrices that included chicken sausages, beef balls, and egg-milk biscuits. The recoveries ranged from 91% to 109% with qualified relative standard deviations <15% with the limit of quantification (of 1.6 mg kg , TM/matrix).
This new approach can be used for the qualitative and quantitative detection of crustacean TM in various food matrices. © 2021 Society of Chemical Industry.
由于甲壳类产品消费的增加，人们对其产生过敏反应的情况也越来越多。肌球蛋白（TM）是主要的甲壳类过敏原，它具有卷曲螺旋结构，对各种食品加工方法都具有稳定性。甲壳类加工产品已被应用于多种食品中，这使得在这些产品中检测 TM 变得更加困难。我们旨在开发一种基于高效液相色谱-串联质谱的方法，用于准确、可重现地定量检测食品中的甲壳类 TM。
选择了三个肽段 IQLLEEDLER、LAEASQAADESER 和 IVELEEELR 作为肽标记物，并选择肽段 IVELEEELR 作为定量标记物。完全优化了提取条件和酶解条件。在 Tris-盐酸缓冲液（50mmol/L，pH7.4）中加入 1mol/L 氯化钾的提取溶液，以及酶/蛋白比（m/m）为 1:15 的酶解处理 13h，效果极佳。该方法在 1 至 1000μg/L 宽范围内表现出良好的线性关系，合格的决定系数（R2=0.9994）。在空白基质（鸡肉香肠、牛肉丸和蛋奶饼干）中三个加标水平（12.5、25.0 和 50.0μg/kg，TM/基质）下进行的准确度验证表明，回收率在 91%至 109%之间，相对标准偏差<15%，定量限（TM/基质为 1.6mg/kg）合格。
该新方法可用于定性和定量检测各种食品基质中的甲壳类 TM。© 2021 化学工业协会。
