Absolute quantitation of β-lactoglobulin by protein liquid chromatography -: Mass spectrometry and its application to different milk products

被引:45
作者
Czerwenka, Christoph
Maier, Irene
Potocnik, Natascha
Pittner, Fritz
Lindner, Wolfgang
机构
[1] Univ Vienna, Inst Analyt Chem & Food Chem, A-1090 Vienna, Austria
[2] Univ Vienna, Dept Biochem, A-1030 Vienna, Austria
关键词
D O I
10.1021/ac062367d
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The absolute quantitation of proteins in biological matrixes is of great interest in many fields and can be accomplished by different methodologies. Here, a method for the absolute quantitation of the whey protein beta-lactoglobulin using protein liquid chromatography coupled to mass spectrometry is reported. The developed approach was characterized in detail and applied to the determination of beta-lactoglobulin contents in various milk products. A special focus was placed on the recovery rates of the isolation procedure and on robust quantitation by LC-MS. For these purposes protein internal standards were employed. The observed recovery rates of beta-lactoglobulin from various samples ranged from 100% for whole milk to just over 50% for a strongly processed yogurt-based baby food product. The influence of processing was investigated in greater detail, showing that an increasing intensity of the applied heat treatment resulted in an increasing loss of beta-lactoglobulin. LC-MS quantitation at the protein level proved to be highly suitable, avoiding a potentially problematic digestion step. The use of an appropriate internal standard to compensate for sample losses during sample workup was shown to be essential for obtaining accurate results.
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页码:5165 / 5172
页数:8
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