Abstract
The quality of food is influenced by several factors during production and storage. When using marker compounds, different steps in the production chain, as well as during storage, can be monitored. This might enable an optimum prediction of food's shelf life and avoid food waste. Especially, proteoforms and peptides thereof can serve as indicators for exogenous influences. The development of a proteomics-based workflow for detecting and identifying differences in the proteome is complex and time-consuming. The aim of the study was to develop a fast and universal workflow with ultra-high temperature (UHT) milk as a proteinaceous model food with expectable changes in protein/peptide composition. To find an optimum shelf life without sticking to a theoretically fixed best-before date, new evaluation and analytical methods are needed. Consequently, a modeling approach was used to monitor the shelf life of the milk after it was treated thermally and stored. The different peptide profiles determined with high-resolution mass spectrometry (HRMS) showed a significant difference depending on the preparation method of the samples. Potential marker peptides were determined using orthogonal projections to latent structures discriminant analysis (OPLSDA) and principal component analysis (PCA) following a typical proteomics protocol with tryptic hydrolysis. An additional Python-based algorithm enabled the identification of eight potential tryptic marker peptides (with mass spectrometric structural indications m/z 885.4843, m/z 639.3500, m/z 635.8622, m/z 634.3570, m/z 412.7191, m/z 623.2967, m/z 880.4767, and m/z 692.4041), indicating the effect of the heat treatment. The developed workflow is flexible and can be easily adapted to different research questions in the field of peptide analysis. In particular, the process of feature identification can be carried out with significantly less effort than with conventional methods.
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