1H NMR for Comparative Metabolic Analysis of Whey and WPC-80

Background/Objectives: Metabolites are low-molecular-weight organic compounds (<1 kDa) that act as intermediates and end products of cellular metabolism. Their characterization provides valuable information on the nutritional quality, functionality, and potential health impacts of food products. In the dairy sector, proton nuclear magnetic resonance (H-1 NMR) spectroscopy has emerged as a powerful tool for metabolite profiling, enabling the simultaneous identification and quantification of diverse compounds. In this study, H-1 NMR was applied to characterize and compare the metabolic composition of whey, a major by-product of cheese and yogurt production, and whey protein concentrate (WPC-80), a whey derivative containing approximately 80% protein by weight and rich in essential amino acids. Methods: Five whey and four WPC-80 samples from a single Parmigiano Reggiano dairy plant were collected, each representing a biologically independent sample. Statistical evaluation was performed using Mann-Whitney U tests to identify significantly different metabolites between groups, while principal component analysis and partial least squares discriminant analysis were employed to assess group separation and determine discriminant metabolites. Results: The results revealed marked compositional differences: whey was higher in dimethyl sulfone, succinate, orotate, fumarate, and lactose (p < 0.05), whereas WPC-80 contained significantly higher levels of histidine, formate, glucose + glucose-6-phosphate, acetate, and choline (p < 0.05). Moreover, metabolites such as hippurate, valine, lactate + threonine, and uracil were exclusively found on whey and not in WPC-80, likely due to processing steps such as ultrafiltration. Conclusions: These findings highlight the metabolic distinctions introduced by WPC-80 processing from Parmigiano Reggiano whey and provide insights into the nutritional and functional characteristics of whey-derived products. Such knowledge can inform the design of innovative dairy ingredients and functional foods, with potential benefits for both industry applications and consumer health.