Absolute Quantitation of beta-Lactoglobulin by Protein Liquid Chromatography-Mass Spectrometry and Its Application to Different Milk Products

摘要

The absolute quantitation of proteins in biological matrixesis 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 100percent for whole milk to just over 50percent 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.