建立并验证了一种反相高效液相色谱二极管阵列检测法，用于同时定量检测乳清蛋白中的微量和主要成分。
Development and validation of an RP-HPLC DAD method for the simultaneous quantification of minor and major whey proteins.
机构信息
University of Hohenheim, Institute of Food Science and Biotechnology, Department of Soft Matter Science and Dairy Technology, Garbenstrasse 21, 70599 Stuttgart, Germany.
University of Hohenheim, Institute of Food Science and Biotechnology, Department of Soft Matter Science and Dairy Technology, Garbenstrasse 21, 70599 Stuttgart, Germany.
出版信息
Food Chem. 2021 Apr 16;342:128176. doi: 10.1016/j.foodchem.2020.128176. Epub 2020 Sep 28.
Whey represents a valuable protein source for human nutrition. Whey composition varies with respect to process characteristics during milk processing. For efficient exploitation of this dairy side stream, reliable analytical methods are essential. The aim of this study was to develop and validate an RP-HPLC-DAD method for the simultaneous quantification of the minor (lactoferrin, lactoperoxidase, bovine serum albumin) and major (α-lactalbumin, β-lactoglobulin) whey proteins. Seven RP-columns were compared and the composition of the mobile phase was optimized to achieve baseline separation. In validation experiments the limits of detection (LOD < 8 mg/L) and quantification (LOQ < 24 mg/L) were determined. Validity was proofed by precision (>96%), accuracy (95% - 103%) and recovery (96% - 102%) measurements. Peak homogeneity was confirmed by SDS-PAGE. The individual working ranges were adjusted to the estimated protein concentrations in whey, allowing direct analysis without sample preparation at a method runtime of 23 min.
乳清代表了一种有价值的人类营养蛋白质来源。乳清的组成因牛奶加工过程中的工艺特点而异。为了有效利用这种乳制品副产物，需要可靠的分析方法。本研究旨在开发和验证一种用于同时定量测定微量（乳铁蛋白、乳过氧化物酶、牛血清白蛋白）和主要（α-乳白蛋白、β-乳球蛋白）乳清蛋白的反相高效液相色谱-二极管阵列检测（RP-HPLC-DAD）方法。比较了七种 RP 柱，并优化了流动相的组成以实现基线分离。在验证实验中，确定了检测限（LOD < 8 mg/L）和定量限（LOQ < 24 mg/L）。通过精密度（>96%）、准确度（95%-103%）和回收率（96%-102%）测量证明了方法的有效性。通过 SDS-PAGE 确认了峰的均一性。根据乳清中估计的蛋白质浓度调整了各个工作范围，允许直接分析，而无需样品制备，方法运行时间为 23 分钟。
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