建立了16 min内检测牛奶中β-酪蛋白、αS1-酪蛋白、αS2-酪蛋白、κ-酪蛋白、α-乳白蛋白和β-乳球蛋白6种主要蛋白质的超高效液相色谱-四极杆飞行时间质谱定量方法。牛奶样品经前处理后,用C18色谱柱分离,四级杆飞行时间质谱系统检测,外标法定量,并对不同蛋白质的亚型进行了解析。结果表明,6种蛋白质在各自浓度范围内呈现良好的线性关系(R2>0.99), 3个不同添加水平的加标回收率为92.43%~107.69%,保留时间日内精密度和日间精密度分别为0.14%~0.39%和0.17%~0.37%,峰面积日内精密度和日间精密度分别为3.06%~8.07%和3.90%~9.42%。方法在实现6种主要蛋白质定量检测的同时,有望对乳制品品质分析以及品种鉴定提供技术支撑。
A quantitative method of ultra-performance liquid chromatography-quadrupole/ time of flight-mass spectrometry for the determination of β-casein, αS1-casein, αS2-casein, κ-casein, α-lactalbumin and β-lactoglobulin in milk within 16 minutes was established. After sample pretreatment, milk from four different provinces was separated by Agilent ZORBAX 300Extend-C18(2.1 mm×100 mm, 300 A) chromatographic column, and finally detected in a mass spectrometry system. The quantitative method was the external standard method, and the protein subtypes were analyzed by Bioconfirm software. The standard curves of six main proteins in milk showed a good linear relationship(R2>0.99), the recovery was 88.52%-105.79%, the relative standard deviation of retention time was 0.14%-0.39%, and the relative standard deviation of peak area was 3.13%-9.42%. The experiment can meet the actual quantitative requirements.
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