STUDY OF METHODOLOGIES FOR EXTRACTION OF PHENOLIC COMPOUNDS IN EXTRA VIRGIN OLIVE OIL FOR CHARACTERIZATION BY HPLC-MS

Olive oil is a type of healthy fat that is often used in the Mediterranean diet due to its antioxidant properties. As this is the most commonly used type of fat in Portuguese cuisine, approximately 230 million liters of olive oil were produced in 2021. Phenolic compounds are among the main components present in olive oil, and they are value-added products due to their potential application in the food, cosmetic, and pharmaceutical industries. The present study aimed to test methodologies for the extraction of phenolic compounds for characterization by HPLC-MS (High-Performance Liquid Chromatography – Mass Spectrometry). To do this, phenolic compounds from extra virgin olive oil produced in Portugal were analyzed using the liquid-liquid extraction technique with the assistance of a centrifuge, employing two different organic solvents. The analysis was performed using the Thermo Fisher Scientific Dionex Ultimate 3000 HPLC system (USA) coupled with an LCQFleet mass spectrometer and a reverse-phase HyperSep C18 column (150mm x 5 x 2.5 μm) from Thermo Scientific. The analysis conditions were chosen according to the type of sample. The first extraction was carried out using n-hexane and ethyl acetate, while the second one employed n-hexane, methanol (MeOH), and Milli-Q water. N-hexane was used as the first wash to remove the lipid phase, and the MeOH: H2O (60:40) solution was used to separate the phase of interest. The best results were obtained through the second assay, identifying some phenols from the Secoiridoid group. The results obtained are in accordance with those found in the literature, demonstrating the presence of compounds beneficial for consumer health, confirming the importance of olive oil in the Mediterranean diet.