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Background: The composition of extra virgin olive oil (EVOO) defines its sensory, nutritional, and human health benefits, and distinguishes it as a key component of the Mediterranean diet. Nevertheless, EVOO constituents are susceptible to degradation during processing and storage, which reduces the olive oil's quality and limits its shelf life. The present study investigated the effect of molecular filtration before storage and the effect of cool storage at 4 °C on the stability of 'Kolovi' EVOO, a variety originating from the Greek island of Lesvos, over a 24 month period., Results: Storing EVOO at 4 °C positively affected free acidity, peroxide value, K 268 , fruity qualities, and concentrations of hydroxytyrosol, tyrosol, ligstroside aglycone, lutein, and squalene, in comparison with the control sample stored at room temperature, particularly after 1 year. Molecular filtration significantly affected the ratio of unsaturated fatty acids to saturated fatty acids (UFAs/SFAs). Optimal preservation of parameters such as acidity value and lutein content was achieved by combining molecular filtration with refrigeration., Conclusions: The present study recommends storing EVOO in the refrigerator for up to 18 months. Based on the regulatory limits of the quality characteristics of acidity, peroxide value, K 232 value and fruity sensory attributes, the shelf-life of the protected geographical indication (PGI) 'Kolovi' EVOO can reach 2 years under cool storage (4 °C) and with molecular filtration before storage. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)