For the first time, High-Resolution Magic Angle Nuclear Magnetic Resonance spectroscopy (NMR-HRMAS) was applied to directly identify specific metabolites from a Spanish raw ewe’s milk and enzymatic coagulation pressed-curd cheese (Protected Geographical Indication: Castellano) manufactured by two procedures (traditional/artisanal vs. industrial) and including the ewe’s raw milk. The NMR parameters were optimized to study the complex matrixes of this type of cheese. In addition, conventional overcrowded ¹H-NMR-HRMAS spectra were selectively simplified by a Carr–Purcell–Meiboom–Gill (CPMG) sequence or a stimulated echo pulse sequence by bipolar gradients (DIFF), thus modulating spin–spin relaxation times and diffusion of molecular components, respectively. ¹H-NMR-HRMAS spectroscopy displayed important information about cheese metabolites, which can be associated with different manufacturing processes (industrial vs. traditional) and ripening times (from 2 to 90 days). These results support that this spectroscopy is a useful technique to monitor the ripening process, from raw milk to commercial ripened cheese, using a minimum intact sample, implying the absence of time-consuming sample pretreatments.

The evolution of structural changes and the textural features during the ripening process of four varieties of Spanish sheep cheese were studied using Magnetic Resonance Imaging (MRI). Specifically, longitudinal (T₁) and transverse (T₂) relaxation times and apparent diffusion coefficient maps were analyzed. Also, proton density was used to improve the description of the structure of the cheeses. The MRI results displayed important information about cheese matrix structure, associated with different manufacturing processes (industrial vs. traditional), ripening times (RTs, from 2 to 180 days), and geographical origins. A significant interaction between RT and cheese variety related to the variations in physicochemical and textural parameters was found. Linear regression models were developed per the abundant literature. Logarithmic regression models showed the highest R² when monitoring the dependency on T₁ and T₂ parameters of water content, water activity, RT, and some texture parameters. Therefore, these results support that MRI is a useful technology to monitor the ripening process, predict textural behavior and physicochemical variables, and characterize the structure of different varieties of sheep cheese.

A comprehensive characterization of two pressed-curd cheeses produced from ewe’s milk using enzymatic coagulation—Manchego cheese (with Protected Designation of Origin, PDO) and Castellano cheese (with Protected Geographical Indication, PGI)—was performed throughout the manufacturing process (industrial or traditional) and ripening stages (2, 9, 30, 90, and 180 days). Proton high-resolution magic angle spinning nuclear magnetic resonance (¹H HRMAS NMR) spectroscopy, combined with Principal Component Analysis (PCA) and cluster analysis, was applied to intact cheese samples. The combination of this spectroscopic technique with chemometric methods allows for the characterization of each type of sheep milk cheese according to its geographical origin and production method (artisanal or industrial), as well as the estimation of ripening time. The results demonstrate that HRMAS NMR spectroscopy enables the rapid and direct analysis of cheese samples, providing a comprehensive profile of their metabolites—a metabolic ‘fingerprint’.