¹H-NMR Spectroscopy and Chemometric Fingerprinting for the Authentication of Organic Extra Virgin Olive Oils

The authentication of organic extra virgin olive oils (OEVOOs) is crucial for quality control and fraud prevention. This study applies proton-nuclear magnetic resonance (¹H-NMR) spectroscopy combined with chemometric analysis as a non-destructive, untargeted approach to differentiate EVOOs based on cultivation method (organic vs. conventional) and variety (<i>Hojiblanca</i> vs. <i>Picual</i>). Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) demonstrated well-defined sample differentiation, while the variable importance in projection (VIP) selection and Tukey’s test identified key spectral regions responsible for classification. The results showed that sterols and lipid-related compounds played a major role in distinguishing organic from conventional oils, whereas fatty acids and phenolic compounds were more relevant for cultivar differentiation. These findings align with known metabolic differences, where <i>Picual</i> oils generally exhibit higher polyphenol content, and a distinct fatty acid composition compared to <i>Hojiblanca</i>. The agreement between chemometric classification models and statistical tests supports the potential of ¹H-NMR for OEVOO authentication. This method provides a comprehensive and reproducible metabolic fingerprint, enabling differentiation based on both agronomic practices and genetic factors. These findings suggest that ¹H-NMR spectroscopy, coupled with multivariate analysis, could be a valuable tool for quality control and fraud detection in the olive oil industry.