Abstract

This study presents a comprehensive metabolomic analysis of Parmigiano Reggiano samples to differentiate between those designated as Mountain Quality Certification (QC) and conventional Protected Designation of Origin (PDO). Despite following the same production protocol, these cheese varieties differ in the cows’ feeding regimes and milk stable locations, with mountain-certified samples adhering to specific requirements regarding milk origin and feed composition. An untargeted approach with Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) was proposed to characterize the cheese metabolome. High-resolution LC-MS data can generate gigabyte-sized files, making data compression essential for manageable multivariate analysis and noise reduction. This study employs the Region of Interest-Multivariate Curve Resolution (ROI-MCR) protocol to achieve effective data compression and chromatographic resolution, thereby extracting the most informative features. This method was compared with a classical approach for feature extraction from chromatographic data, namely Compound Discoverer (CD) software. The features extracted by both methods were analysed through Principal Component Analysis (PCA) and ASCA (ANOVA Simultaneous Component Analysis). The comparison of ROI-MCR and CD approaches demonstrated that while both methods yielded similar overall conclusions, ROI-MCR provided a more streamlined and manageable dataset, facilitating easier interpretation of the metabolic differences. Both approaches indicated that amino acids, fatty acids, and bacterial activity-related compounds played significant roles in distinguishing between the two sample types.

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