Departamento de Ciencia y Tecnología de los Alimentos, Facultad de Farmacia, Universidad de Concepción , Concepción, Chile.
Laboratorio de Biotecnología y Genética de Alimentos, Departamento de Ciencia y Tecnología de Los Alimentos, Facultad de Farmacia, Universidad de Concepción , Concepción, Chile.
Food Addit Contam Part B Surveill. 2020 Dec;13(4):268-274. doi: 10.1080/19393210.2020.1769194. Epub 2020 May 25.
Casein and ovalbumin are proteins commonly used as wine fining agents that may trigger allergic reactions in susceptible individuals. Therefore, their occurrence in wines could become a health risk, moreover when their presence is not reported. The objective of this work was to determine the occurrence of casein and ovalbumin in Chilean wines. Proteins were extracted by ultrafiltration and digested applying an optimised ultrasound-assisted method. Peptides were quantified by validated tandem mass spectrometry method using stable isotope dilution analysis. Optimal digestion was achieved in 3 minutes at a 1:10 enzyme protein ratio. The method was validated following ICH guidelines showing determination coefficients R ≥ 0.99, repeatability, and intermediate precision with RSD values <1.95% and recoveries from 89.8% to 115.1% (RSD < 5.84%). The method was applied to analyse 60 wine samples. Fifty-six samples showed quantifiable levels, from which 28% presented a total casein and ovalbumin concentration equal or higher to the European limit (0.25 mg L).
酪蛋白和卵清蛋白是常用的葡萄酒澄清剂蛋白，它们可能会引发易过敏个体的过敏反应。因此，如果这些物质出现在葡萄酒中而未被报告，可能会对健康造成风险。本研究旨在检测智利葡萄酒中是否存在酪蛋白和卵清蛋白。采用超滤法提取蛋白，采用优化后的超声辅助法进行酶解。利用稳定同位素稀释分析，通过经验证的串联质谱法对肽段进行定量分析。在酶蛋白比为 1:10、3 分钟的条件下可实现最佳酶解。方法通过 ICH 指南验证，结果表明，决定系数 R≥0.99，重复性和中间精密度的相对标准偏差（RSD）值均<1.95%，回收率在 89.8%至 115.1%之间（RSD<5.84%）。该方法应用于 60 个葡萄酒样品的分析，其中 56 个样品可定量检测到蛋白，28%的样品总酪蛋白和卵清蛋白浓度等于或高于欧洲限量（0.25mg/L）。
