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The fatty acids of olive oils of distinct quality grade from the most important European Union (EU) producer countries were chemically and isotopically characterized. The analytical approach utilized combined capillary column gas chromatography−mass spectrometry (GC/MS) and the novel technique of compound-specific isotope analysis (CSIA) through gas chromatography coupled to a stable isotope ratio mass spectrometer (IRMS) via a combustion (C) interface (GC/C/IRMS). This approach provides further insights into the control of the purity and geographical origin of oils sold as cold-pressed extra virgin olive oil with certified origin appellation. The results indicate that substantial enrichment in heavy carbon isotope (13C) of the bulk oil and of individual fatty acids are related to (1) a thermally induced degradation due to deodorization or steam washing of the olive oils and (2) the potential blend with refined olive oil or other vegetable oils. The interpretation of the data is based on principal component analysis of the fatty acids concentrations and isotopic data (δ13Coil, δ13C16:0, δ13C18:1) and on the δ13C16:0 vs δ13C18:1 covariations. The differences in the δ13C values of palmitic and oleic acids are discussed in terms of biosynthesis of these acids in the plant tissue and admixture of distinct oils. Keywords: Olive oil; carbon isotope; isotopic characterization; fatty acids; adulteration