Stability of refined olive oil and olive‐pomace oil added by phenolic compounds from olive leaves

Refined olive oil and olive‐pomace oil were enriched with olive leaf phenolic compounds in order to enhance its quality and bring it closer to virgin olive oil. The changes that occurred in the concentrations of pure oleuropein, oleuropein aglycone, hydroxytyrosol acetyl and α‐tocopherol at 400 µg/kg of oil during the storage of refined olive oil and olive‐pomace oil under accelerated conditions (50 °C) were investigated. In a period of 4 months, α‐tocopherol decomposed by 75% whereas less than 40% of the phenols were lost. During storage, enzymatic olive leaf extract hydrolysate that contains two major compounds, hydroxytyrosol and oleuropein aglycone showed the highest antioxidant activity and the lowest detected stability, followed by oleuropein. The oleuropein in olive leaf extracts exhibited similar degradation profiles, reducing by 60–50% and 80% for the olive oil and olive‐pomace oil in 6 months, respectively. The acetylated extract, however, displayed a loss of 10 and 5% in olive oil and olive‐pomace oil, respectively. In the fatty acid composition, an increase in oleic acid and a decrease in linoleic acid were observed. The antiradical activities of the olive oil and olive‐pomace oil enriched with olive leaf phenolic compounds at 400 ppm showed that enzymatic hydrolysate extract had the highest protective effect against oil oxidation. Based on the Rancimat method, the oils with added leaf enzymatic hydrolysate extract had the lowest peroxide value and the highest stability. After 6 months of storage and at 120 °C, the oxidative resistance of refined olive oil and olive‐pomace oil reached 0.71 and 0.89 h, respectively, whereas that of the non‐enriched samples fell to zero.     

Effect of location on virgin olive oils of the two main Tunisian olive cultivars

The olive oil content in phenolic compounds depends on the variety of the fruit used for its extraction as well as on the predominant climate conditions in the tree cultivation area. Here, we report on the characterization of virgin olive oil samples obtained from fruits of the main Tunisian olive cultivars Chemlali and Chétoui, grown in three different Tunisian locations, Zaghouan (North), Sousse (Center) and Sfax (South). Chétoui olive oil samples obtained from fruits of olive trees cultivated in Zaghouan and Chemlali olive oil samples obtained from fruits of olive trees cultivated in Sousse were found to have a higher mean total phenol content (1004 and 330 mg/kg, respectively). Olive oil samples obtained from fruits of both cultivars had different phenolic profiles and a higher content in 3,4‐DHPEA‐EDA when the olive trees were cultivated in Zaghouan. Both olive cultivars were found to have different responses to environmental conditions. Chétoui olive oil showed decreased oxidative stability when the fruits were obtained from olive trees cultivated in the center of Tunisia (34.8 h) and in Sfax (16.17 h). Furthermore, statistical data showed that the phenolic composition and oxidative stability of Chétoui olive oil varied more by location than those of Chemlali olive oils.     

Natural antioxidants and volatile compounds of virgin olive oils obtained by two or three‐phases centrifugal decanters

Research has been carried out to ascertain the influence of different centrifugal decanters employed in olive process on oil yields and qualitative characteristics and composition of volatile compounds of virgin olive oil. Tests were performed in an olive oil mill equipped with centrifugal decanters at two or three‐phases. Results show that oil yields were similar and oils extracted from good‐quality olives do not differ in free fatty acids, peroxide value, UV absorptions and organoleptic assessment. Total phenols and o‐diphenols content as well as induction time values are higher in oils obtained by the centrifugal decanter at two‐phases, because it requires less quantity of water added to olive paste in comparison to the three‐phases centrifugal decanter. The amount of water added determines the dilution of the aqueous phase and lowers the concentration of the phenolic substances more soluble in vegetable waste water. Due to the partition equilibrium law the concentration of the same substances consequently diminishes in the oil. In this research, the coefficient of the partition equilibrium of total phenols between oil and vegetable water has been calculated and discussed. No significant difference occurred, due to the different decanters employed, in the average values of the volatile components of the head‐space of oils.     

Enrichment of pomace olive oil in triterpenic acids during storage of “Alpeorujo” olive paste

Triterpenic acids are natural compounds present in plants and foods with beneficial properties for human health and thus they are desirable in the food, cosmetics and pharmaceutical industries. Pomace olive is considered a good source of these substances. In this study, oleanolic and maslinic acids were found to be the main triterpenic acids identified in pomace olive oil obtained from stored “Alpeorujo”. Determination of the two acids was achieved by a new procedure that consists of extracting the acids from the oil with a mixture of methanol/ethanol, and then separating and quantifying them by HPLC. Results showed that their concentration increased up to 16 g/kg of oil during storage of the pomace in large ponds for 7 months. The concentration of both triterpenic acids was similar in the pomace olive oil obtained by using the centrifugation system. By contrast, a much lower concentration of maslinic than oleanolic acid was detected in pomace olive oils obtained by solvent extraction from the previously centrifugated “Alpeorujo” paste. These triterpenic acids also contributed to the acidity of the crude oil. Likewise, the oil of the pomace paste was enriched in other substances such as 4‐ethylphenol and aliphatic alcohols during the storage of the paste in large ponds. Consequently, crude pomace olive oil can be considered a good source of triterpenic acids when obtained from a stored olive paste.     

Influence of olive crushing methods on the yields and oil characteristics

In the last years, metallic crushers substituted granite stone mill with some variations in the organoleptic oil characteristics. To control the influence of the crushing method on the yield and oil quality, the olive pastes were obtained using three different ways: (i) new metallic crusher at mobile knives; (ii) granite stone mill; (iii) double olive crushing by the metallic crusher and the granite stone mill. With the aim to ascertain the useful use of a new metallic crusher (at mobile knives), experimental tests were carried out in an industrial oil mill. This oil mill is equipped by a centrifugal decanter generating two oil flows: first and second extraction (recovery) oils. The results showed that the yields obtained by different methods were satisfactory. No statistically significant differences have been observed in terms of oil yield and quality when different crushing devices were used. All first extracted oils are extra virgin with similar organoleptic characteristics, especially for the fruity intensity and for the bitter and pungent taste, as confirmed by the composition of volatile substances and the content of phenolic oil compounds. The recovery oils (second extraction oils) showed, in contrast to first extraction oils, a more intense green colour and a higher content of total phenols. Practical applications: Processing of sound olives with the right ripening grade and good quality allows to easily obtain an extra virgin olive oil, with commercial qualitative parameters according to the European Union requirements. However, different olive crushing systems affect the concentrations of some compounds responsible of aroma and taste (phenolic compounds). The use of the more violent metallic crushers facilitates obtaining oils with total phenol content higher than when using a stone mill. Here we used a particular metallic crusher (at knives) that, however, is suitable to replace the granite stone mill when a less pungent and bitter oil is required.     

Stability of a phenol‐enriched olive oil during storage

The use of an emulsifier to stabilize the phenolic compounds added in the preparation of an enriched olive oil was evaluated. Two emulsifiers, lecithin and monoglyceride, were studied. The results showed lecithin to be the most convenient, due to the increase in the value of the oxidative stability of the phenol‐enriched oils in relation to the enrichments prepared with monoglycerides. After that, the shelf life of the prepared oils was evaluated during a period of 256 days of storage at 25°C in the dark. Oil quality parameters, total phenolic content, bitterness index and oxidative stability were studied during the storage period. Additionally, the phenolic composition and antioxidant capacity (by using the ORAC assay) were evaluated at the end of the storage. The phenolic enrichment of the oils allowed the shelf life of the oils to be extended compared with the control (virgin olive oil without phenol addition), delaying the appearance of peroxides and improving their oxidative stability. In addition, the higher content of phenolic compounds in the oils at all stages of storage is desirable in order to increase the intake of these beneficial compounds. Practical applications : The preparation of phenol‐enriched olive oils with a higher phenolic content than the commercial virgin olive oils is of special interest to increase the ingestion of these healthy compounds the daily intake of which is limited due to the high caloric value of olive oil. There are two key points in the development of this product: (i) the dispersion and stabilization of the phenol extract in the oil matrix and (ii) the stability of the phenols in the prepared oils to guarantee the phenol concentration during their shelf life. It is important to study the use of emulsifiers to determine if they allow an improvement in the dispersion of the phenolic extract, and their stabilization in the final product. In addition, the emulsifiers could mask the bitter taste of the enriched oils, which is desirable to increase consumer acceptance of the enriched oil.     

Optimization of the Abencor system to extract olive oil from irrigated orchards

Studying the composition of olive oil requires cold‐press olive oil extraction. One of the most common laboratorial mills is the Abencor system. However, its operation protocol was formulated decades ago for Spanish olive varieties from traditionally rain‐fed orchards. We modified this protocol for use with “Barnea” and “Picual” olives from irrigated orchards that are characterized by high water content. Independent effects of malaxation time, temperature, water addition and talc addition on extraction efficiency, and major quality indices of virgin olive oil were studied. Overall, addition of talc to the fruit paste was the most significant treatment in terms of yield and quality of the oil although its effect was cultivar dependent. Improved oil yield was particularly significant for “Picual.” Extended malaxation time was also effective in improving oil extractability. Addition of talc generally improved oil‐quality parameters, while water addition had the opposite effect. Yet, quality parameters remained within the extra virgin level. Temperature increments reduced oil quality. The need to adapt a modified protocol for use with fruits from irrigated orchards that will facilitate critical comparison of results obtained from different agronomic theses and different laboratories is highlighted. It is recommended that each laboratory develops an appropriate protocol for the operation of the Abencor system in accordance to the characteristics of the olive fruit they are working with. Practical applications: Abencor system serves as the major laboratorial mill world‐wide. Those mills allow the researchers to characterize olive oil in accordance to the treatments received by the trees. This cannot be done in commercial mills. The system operation protocol was established decades ago for fruits from rain‐fed orchards. In the past decade there was a rapid increase in the use of irrigation in olive orchards and therefore it is crucial to optimize the operation protocol for fruit with relatively high water content. In the current work we have evaluated the influence of a series of technological parameters (i.e., talc and water addition, malaxation time, and temperature) on the extraction efficiency and quality indices of olive oil. This allowed us to present a modified protocol for the Abencor system operation suitable for olive fruit of irrigated orchards that will facilitate a reliable representation of the influence of different treatments on the yield and characteristics of the olive oil.     

Evolution of phenols and pigments in extra virgin olive oil from irrigated super‐intensive orchard

Phenolic compounds have a high importance in olive oil because of their effect on shelf life and sensory properties. This study reports on the HPLC profiles of the phenolic compounds of virgin olive oils obtained from Arbequina olives from the harvesting in a super‐intensive orchard under a linear irrigation system. In addition, phenolic content, carotenoid and chlorophyllic pigments, and oxidative stability were analyzed. Total phenol content and 3,4‐DHPEA‐EDA increased up to a maximum throughout the ripening process. The simple phenols tyrosol and hydroxytyrosol acetate increased throughout the ripening process, however, there was not found a clear trend in hydroxytyrosol content. Minor constituents such as vanillic acid and p‐coumaric acid increased up to a maximum and then decreased, since vanillin decreased progressively throughout the time of harvest. 3,4‐DHPEA‐EDA and lignans were present in considerable amounts in the studied samples, while oleuropein aglycone was present in a low amount. Total phenol content and oil stability followed the same trend throughout the study, so a very good correlation was established between them. Total secoiridoids and, specifically, 3,4‐DHPEA‐EDA seemed to be responsible for oil stability. The pigment content decreased during ripening, and not a positive correlation was found between pigments and oil stability. Practical applications : The results can be used to determine the best time for harvesting in order to obtain olive oils with different phenols and pigment contents. This is important for sensory characteristics of the olive oils and also for olive oil stability.     

Effect of olive fruit freezing on oxidative stability of virgin olive oil

Several studies have suggested that the phenolic fraction plays an important role during storage and therefore in the shelf life of virgin olive oil. This investigation examines the effect of freezing olives (–18 °C) before processing into oil on the transfer of the phenolic compounds into the subsequent oil, and the consequential changes in oxidative stability. Oil samples obtained from frozen olives (24 h at –18 °C), crushed with and without preliminary thawing, were compared to a control sample; both oils were obtained using a two‐phase low‐scale mill. The oxidative stability in different samples was assessed in terms of primary and secondary oxidation products as measured by peroxide values and oxidative stability index times, respectively. The quality of the oil samples was also checked through the percentage of free acidity and the phenolic content. Phenols were determined by both spectrophotometric assays (total phenols and o‐diphenols) and HPLC‐DAD/MSD. The antiradical capacity of the phenolic fraction was determined by DPPH and ABTS spectrophotometric tests. These analyses showed that thawing of olives before oil extraction led to a significant loss of oxidative stability and phenols; in contrast, samples obtained from frozen olives that were not thawed before crushing showed qualitative characteristics similar to control samples.     

Understanding degradation of phenolic compounds during olive oil processing by inhibitor addition

The aim of this work was to study, under different conditions, degradation of secoiridoids during extraction of extra virgin olive oil by following the effect of ascorbic and citric acid addition. Their effect was evaluated on oil obtained from both damaged olives and undamaged fresh olives. Addition of enzyme inhibitors to damaged olives during olive milling allowed us to obtain oil with a higher phenolic compound content. Conversely, addition of the same inhibitors to undamaged fresh olives, during oil milling, resulted in no significant improvement in the phenolic compound content of oil. A high presence of PPO was thus indirectly confirmed, as damaged olives were only found to be sensitive to action of inhibitors. Ascorbic acid was found to be more effective than citric acid in preserving phenolic compounds of oil. Trials on undamaged fresh olives confirmed occurrence of hydrolytic transformation phenomena for secoiridoids during extra virgin olive oil production process. In particular, the quantitatively most representative component for Frantoio cultivar was found to be 3,4‐DHPEA‐EDA. This compound may be considered a direct marker for the degree of transformation of secoiridoids during production process. Practical applications: The processing of undamaged olives resulted in the extraction of extra virgin olive oil with a higher phenol content. It could be indirectly inferred that a reduced activity of PPO caused a low secoiridoid degradation both before and after malaxation. Lightly scratched, overripe olives could be used in those markets where the addition of oxidation‐inhibiting substances is allowed. Using inhibitors can be suggested for olive washing step.     

Phenolic composition of virgin olive oils from cross breeding segregating populations

The evaluation and characterization of segregating populations is a critical step in olive breeding programs. In this work, phenolic profiles of virgin olive oils (VOOs) from segregating populations obtained by cross breeding in Cordoba (Spain) have been evaluated. Genotypes obtained from open pollination of the cultivar Manzanilla de Sevilla, and from crosses between the cultivars Arbequina × Arbosana, Picual × Koroneiki and Sikitita × Arbosana were tested. The phenolic composition was determined after liquid–liquid extraction with 60:40 v/v methanol–water and subsequent chromatographic analysis with ultraviolet (UV) detection of both absorption and fluorescence in a sequential configuration. Results for all studied compounds showed high degree of variability between genotypes, with a higher range of variation than the observed for the genitors. Most of the observed variability was attributable to differences in genotypes within crosses rather than among crosses. Some issues related to breeding strategies are discussed. Practical applications: Phenolic compounds are considered to be of paramount importance for the assessment of virgin olive oil quality due to their contribution to the nutraceutical and sensory profile of this natural food. This study focuses on the evaluation of the content of phenolic compounds in olive oils originated from cross breeding in an olive breeding program (Cordoba, Spain). This step is crucial to determine the range of variation of phenolic compounds and the selection of interesting genotypes with higher composition in total phenols or in an individual phenol targeted at a breeding program.     

High-performance liquid chromatography evaluation of phenols in virgin olive oil during extraction at laboratory and industrial scale

Phenolic compounds are of fundamental importance to the quality and nutritional properties of virgin olive oils. In this paper, the high-performance liquid chromatographic analysis of simple and complex olive oil phenols in the streams generated in the two-phase extraction system was carried out using Arbequina and Picual olives. The malaxation stage reduced the concentration of orthodiphenols in oil ca 50–70%, while the concentration of the nonorthodiphenols remained constant, particularly the recently identified lignans 1-acetoxypinoresinol and pinoresinol. Oxidation of orthodiphenols at laboratory scale was avoided by malaxing the paste under a nitrogen atmosphere. Phenolic compounds in the wash water used in the vertical centrifuge were also identified. Hydroxytyrosol, tyrosol, the dialdehydic form of elenolic acid linked to hydroxytyrosol were the most representative phenols in these waters. Hence, phenolic compounds in the wash waters came from both the aqueous and the lipid phases of the decanter oily must.     

Malaxing temperature affects volatile and phenol composition as well as other analytical features of virgin olive oil

Three Italian olive varieties (Caroleo, Leccino and Dritta) were processed by centrifugation in the oil mill. The olive paste was kneaded at 20, 25, 30 and 35 °C. The results achieved revealed that the oil content in green volatiles from lipoxygenase pathway (including C₅ and C₆ compounds and especially unsaturated C₆ aldehydes) decreased progressively as the kneading temperature increased, dropping markedly at 35 °C. The content of phenols, o‐diphenols and secoiridoids showed an opposite trend, but the temperature of 35 °C was critical also for them, as it was for the majority of the other components, analytical parameters and indices related to quality, typicality and genuineness. In general, an increasing kneading temperatures increased the release of oil constituents from the vegetable tissue. This factor also affected the oil extraction yields. The best overall results were achieved by malaxing the olive paste at 30 °C. In fact, this temperature level led to achieving both pleasant green virgin olive oils and satisfactory oil extraction outputs.     

Analytical characteristics of virgin olive oil from young trees (arbequina cultivar) growing under linear irrigation strategies

The effect of a linear (vs. effective crop coefficient, K _c), irrigation strategy applied to young olive trees (Arbequina cv.) on the qualitative and quantitative parameters of virgin olive oil quality was studied. Although linear irrigation strategy did not affect the quality indexes used to classify olive oil by commercial grades, it did influence other important parameters such as total phenol content, bitter index oxidative stability, and the sensorial appraisal. All of these of olive oil qualities were negatively associated with the amount of applied irrigation water. Pigment content of oils determined by carotenoid and chlorophylls also was negatively associated with the amount of water supplied. No consistent relation was found for α-tocopherol and fatty acid content of olive oil in relation to these treatments.     

Evolution of phenolic compounds in virgin olive oil during storage

Phenolic compounds are of fundamental importance to the shelf life of virgin olive oils because of their antioxidative properties. In this paper, the evolution of simple and complex olive oil phenols during 18 mon of storage is studied by high-performance liquid chromatography (HPLC) analysis. The olive oils under examination were from various olive cultivars, harvested in two sectors in the same region at different stages of ripeness. The findings indicate that it is not the variety but rather the ripeness of the olives and the soil and climate that influence the phenol composition of virgin olive oil. In addition, a positive correlation was found between the age of the oils and the tyrosol to total phenols ratio. Lastly, gas chromatography-mass spectrometry analysis confirmed that the unidentified peaks detected by HPLC were of a phenolic nature.     

Influence of Water Deficit in Bioactive Compounds of Olive Paste and Oil Content

The main objective of this study was to evaluate the effect of different deficit irrigation treatments (control, regulated deficit irrigation [RDI]‐1, RDI‐2, and RDI‐3) on the phenolic profile of the olive paste and oil content. Irrigation treatments with more stress water led to a considerable increase in the phenolic compounds of olive paste, especially in oleuropein (60.24%), hydroxytyrosol (82%), tyrosol (195%), and verbascoside (223%) compared to control. A significant increase in the content of total flavonoids and phenolic acids was also observed for these samples. In virgin olive oils (VOO) elaborated from the most stressed olive trees (RDI‐2 and RDI‐3), a noticeable increase in phenolic substances with antioxidant properties (oleuropein, hydroxytyrosol, tyrosol, secoiridoid derivatives, and o‐vanillin) was observed. Consequently, water stress conditions improved antioxidant activity of VOO.     

Phenolic acid content and sensory properties of two Spanish monovarietal virgin olive oils

In recent years, phenolic acids have received considerable attention as they are essential to olive oil quality and nutritional properties. This study aims to validate a rapid and sensitive method based on ultra‐performance liquid chromatography/time‐of‐flight mass spectrometry (UPLC–TOF‐MS) for analyzing the phenolic acid content of olive oil and assessing its impact on virgin olive oil (VOO) sensory attributes. Once this method was validated, we used it to evaluate the phenolic acid composition of several Spanish monovarietal virgin olive oils in relation to nine different olive ripening stages. The results obtained confirm that the methodology developed in this study is valid for extracting and analyzing phenolic acids from VOO. The phenolic acid content of the virgin olive oils sampled was proven to be influenced by the type of cultivar and olive harvest date. Therefore, phenolic acids might be used as potential markers for olive oil cultivar or ripening stage. Finally, the data obtained indicate that the sensory properties of VOO may be differently affected by its phenolic acid content depending on the type of cultivar. Practical applications: The method validated in the present study – based on UPLC‐TOF‐MS – allows experts to assess the phenolic acid content of different VOO cultivars (varieties). This application will probably be very useful to the olive oil industry. The reason is that our study revealed that phenolic acids have an impact on the sensory quality of VOO, which is essential to consumer preferences and choice. In addition, there are phenolic acids that are only found in a particular variety of olive oil obtained from fruits at a specific ripening stage. Consequently, phenolic acids could be used as potential markers for olive oil variety and harvest time.     

On the importance of total polar phenols to monitor the stability of Greek virgin olive oil

A large number of virgin olive oil samples obtained from different areas in Greece were analyzed for various quality parameters. The work focuses on the colorimetric determination of total phenols with the Folin‐Ciocalteu reagent and its importance in predicting shelf life of virgin olive oil. The results indicate a good correlation of total polar phenol content with the stability of the oil. Colorimetric determination of ortho‐diphenol content does not seem to be a better means for predicting virgin olive oil stability. RP‐HPLC quantification of hydroxytyrosol and tyrosol in their free form gives poor results in the case of freshly extracted oils. It is concluded that until an easy‐to‐manage HPLC method will be available, which will quantify accurately both free and bound forms of hydroxytyrosol and other phenolics, the colorimetric method for the determination of total polar phenols remains a good practical means to evaluate the stability of virgin olive oil.     

Hydrophilic antioxidants of virgin olive oil. Part 1: Hydrophilic phenols: A key factor for virgin olive oil quality

Virgin olive oil (VOO) consumption is increasing all over the world due to its excellent organoleptic and nutraceutical properties. These beneficial traits stand from a prominent and well‐balanced chemical composition, which is a blend of major (98% of total oil weight) and minor compounds including antioxidants. The main antioxidants are phenolic compounds, which can be divided into lipophilic and hydrophilic phenols. While lipophilic phenols such as tocopherols can be found in other vegetable oils, most hydrophilic phenols in olive oil are exclusive of the Olea europaea species endowing it with a chemotaxonomic interest. This review is focused on VOO antioxidant profile and, particularly, on hydrophilic phenols that are divided into different sub‐families such as phenolic acids and alcohols, hydroxy‐isochromans, flavonoids, secoiridoids, lignans and pigments. Analytical methods for qualitative and/or quantitative determination of these compounds are assessed. The implementation of efficient sample preparation protocols, separation techniques such as liquid chromatography, GC and capillary electrophoresis, as well as detection techniques such as ultraviolet absorption, fluorescence or MS are critical to succeed in the quality of the results. The effects of hydrophilic phenols on increasing VOO stability, its nutraceutical interest and organoleptic properties are also considered.