Effect of natural antioxidants in virgin olive oil on oxidative stability of refined,bleached, and deodorized olive oil

The factors influencing the oxidative stability of different commercial olive oils were evaluated. Comparisons were made of (i) the oxidative stability of commercial olive oils with that of a refined, bleached, and deodorized (RBD) olive oil, and (ii) the antioxidant activity of a mixture of phenolic compounds extracted from virgin olive oil with that of pure compounds andα-tocopherol added to RBD olive oil. The progress of oxidation at 60°C was followed by measuring both the formation (peroxide value, PV) and the decomposition (hexanal and volatiles) of hydroperoxides. The trends in antioxidant activity were different according to whether PV or hexanal were measured. Although the virgin olive oils contained higher levels of phenolic compounds than did the refined and RBD oils, their oxidative stability was significantly decreased by their high initial PV. Phenolic compounds extracted from virgin olive oils increased the oxidative stability of RBD olive oil. On the basis of PV, the phenol extract had the best antioxidant activity at 50 ppm, as gallic acid equivalents, but on the basis of hexanal formation, better antioxidant activity was observed at 100 and 200 ppm.α-Tocopherol behaved as a prooxidant at high concentrations (>250 ppm) on the basis of PV, but was more effective than the other antioxidants in inhibiting hexanal formation in RBD olive oil.o-Diphenols (caffeic acid) and, to a lesser extent, substitutedo-diphenols (ferulic and vanillic acids), showed better antioxidant activity than monophenols (p- ando-coumaric), based on both PV and hexanal formation. This study emphasizes the need to measure at least two oxidation parameters to better evaluate antioxidants and the oxidative stability of olive oils. The antioxidant effectiveness of phenolic compounds in virgin olive oils can be significantly diminished in oils if their initial PV are too high.     

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.     

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.     

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 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.     

Validation of the oxygen radical absorbance capacity (ORAC) parameter as a new index of quality and stability of virgin olive oil

The induction period (IP), determined using accelerated methods such as the Rancimat test, is a parameter that has been used to predict the shelf life of virgin olive oil. The oxygen radical absorbance capacity (ORAC) has recently been proposed as a quality index of virgin olive oil because it measures the efficiency of phenolic compounds in the protection against peroxyl radicals. This study aims to investigate relationships between the ORAC and IP values and proposes ORAC as a new parameter of virgin olive oil stability. The concentrations of phenolics, o-diphenols, tocopherol, β-carotene, lutein, and ORAC and IP values were determined in 33 virgin olive oils. Regression analyses showed that both ORAC and IP values correlate with total phenols and o-diphenols with highly significant indices, whereas the correlations of both ORAC and IP with tocopherols, β-carotene, and lutein were not significant. The ORAC values correlate with the IP values with low but significant indices (R=0.42; P<0.02). The results confirm the key role of phenolic compounds in accounting for the shelf life of virgin olive oil and suggest that the ORAC parameter may be used as a new index of quality and stability.     

Stabilization of refined olive oil by enrichment with chlorophyll pigments extracted from Chemlali olive leaves

This paper presents the first investigation on the effect of enrichment refined olive oil by chlorophyll pigment extracted from Chemlali olive leaves during storage (6 months). The changes that occurred in the quality indices, fatty acids, sterol, and phenolic content were investigated during the storage of refined olive oil under RT (20°C) and accelerated conditions (50°C) in the dark. Additionally, the pigments (chlorophyll and carotene) changes during 6 months of oil storage were evaluated. At the end of the storage, more than 90% of chlorophyll pigments decomposed in all samples, while, carotene pigment loss was lower showing up to 60 and 85% loss for oil stored at 20 and 50°C, respectively, at the end of storage. The reduction of total phenolic compounds exhibited similar degradation profiles, being reduced by 5% and up to 60% for the enriched refined olive oil stored at 20 and 50°C in 6 months, respectively. In the fatty acid composition, an increase in oleic acid and a decrease in linoleic and linolenic acids were less significant in enriched than non‐enriched refined olive oil. On the other hand, sterol composition was less affected by storage in enriched oil samples. However, the sterol concentration of the oil samples showed an increase in β‐sitosterol, 24‐methylene cholesterol, stigmasterol, and a decrease in cholesterol, Δ5, 24‐stigmastadienol percentage at the end of storage. Based on the Rancimat method, the oils with added leaf pigment extract had the lowest peroxide value and the highest stability. After 6 months of storage, the oxidative resistance of refined olive oil fell to 0.2 and to zero for enriched refined olive oil stored at 20 and 50°C, respectively.     

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.     

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.     

Chemical Composition and Sensory Quality of Tunisian ‘Sayali’ Virgin Olive Oils as Affected by Fruit Ripening: Toward an Appropriate Harvesting Time

Herein, the influence of the ripeness of Tunisian Sayali olives on the chemical composition and sensory quality of virgin olive oils have been investigated, with a particular focus on minor metabolites. Towards this end, five samples (S1–S5) were produced from fruits at increasing stages of maturity and then analyzed. Quality indices (free acidity, peroxide value, specific extinction in UV, sensory characteristics) and composition in major (fatty acids) and minor compounds (squalene, pigments, tocopherols, phenolic compounds, volatile compounds), as well as oxidative stability, were evaluated. Significant variations for the most analytical parameters of Sayali samples were demonstrated, highlighting the impact of stage of ripening. In particular, at later stages of ripening a decreased tendency was seen in minor compounds that are able to inhibit lipid oxidation (tocopherols, carotenoids, squalene and polar phenolic compounds) and, as a consequence, in the oxidative stability value. Moreover, a higher intensity of positive sensory notes (fruity, bitter and pungent) characteristic of extra virgin olive oil were found for samples produced with less ripe olives. Finally, a deeper knowledge of the influence of this factor would be helpful to correctly manage the optimal fruit harvesting time for producers for this variety and to improve the marketing of extra virgin olive oils by using a promising secondary variety (with a high oxidative stability and an interesting fatty acid composition).     

The olive oil oxygen radical absorbance capacity (DPPH assay) as a quality indicator

The antioxidant properties of some single components and the total antioxidant activity of extra‐virgin olive oil have been evaluated by the oxygen radical absorbance capacity (ORAC) method. The total ORAC of the extra‐virgin olive oil was found to be positively correlated with the concentration of total polyphenols, which are important to the shelf life of the product. Among the single phenolic compounds studied, gallic acid showed a higher ORAC than caffeic acid and oleuropein, while among the derivates of oleuropein, hydroxytyrosol was found to be the most active compound among all the phenols studied. The total ORAC of commercial olive oils differed according to the concentration of total polyphenols. The total ORAC of extra‐virgin olive oil was constant during 1 year of storage in rational conditions, whereas it worsened dramatically in olive oil damaged by the lipase‐producing yeast Williopsis californica or by lipase from Pseudomonas spp. The study accomplished on the oily fraction of the fruits before harvesting demonstrated that the total ORAC of the oil of under‐ripe green olives is higher compared to that shown by mature fruits; therefore, through the choice of the harvesting time, it is possible to define also the future content of polyphenols of the oil. The total ORAC test, together with other analyses, can be considered as a qualitative parameter that can contribute to the expression of technological and health virtues of extra‐virgin olive oil.     

Effects of free fatty acids on oxidative stability of vegetable oil

The effect of free fatty acid (FFA) content on the susceptibility to thermooxidative degeneration of vegetable oils was determined by Rancimat analysis. A prooxidant effect of FFA was observed in all filtered oils, independently of lipidic substrate and of its state of hydrolytic and oxidative alteration. The intensity of this effect was related to FFA concentration, but regression analysis of the experimental data did not show a general correlation law between FFA concentration and induction time (I _t). Different results were obtained for freshly processed virgin olive oils, characterized by postpressing natural suspension-dispersion: opposite behavior was observed of FFA content as regards oxidative stability, depending on the presence of suspended-dispersed material. This fact is of interest because the dispersed particles play a double stabilizing effect on both oxidative and hydrolytic degradation. These results showed that avoidance of oil filtration is highly desirable to extend olive oil’s shelf life.     

Analytical evaluation of six monovarietal virgin olive oils from Northern Tunisia

The characterization of virgin olive oils from six Tunisian cultivars, namely Chétoui, Ain Jarboua, Jarboui, Regregui, Rekhami and Neb Jmel, grown in Nebeur (a region of the Kef) was carried out. These cultivars dominate their natural habitats, but with the exception of the Chétoui cultivar they are only scattered throughout the nation. Several analytical parameters were evaluated; these include quality index, fatty acid composition, chlorophylls, carotenoids, sterols, α‐tocopherol and phenolic compounds. Their relationship with oxidative stability was also tested. The main phenols found were tyrosol, hydroxytyrosol, the dialdehydic form of elenolic acid linked to tyrosol and hydroxytyrosol, oleuropein aglycon and pinoresinol. These phenolic compounds, the colorimetric total phenol content and o‐diphenols showed significant correlations with oxidative stability. Furthermore, most of the analytical parameters of the oils that were determined in this study were greatly influenced by genetic factors (cultivar).     

Influence of seasonal conditions on the composition and quality parameters of monovarietal virgin olive oils

The aim of this work was to determine the effect of the climatological conditions of the olive crop season on the composition of monovarietal virgin olive oils obtained from the Arbequina cultivar with special emphasis on the phenolic fraction, its percent distribution, and related oil quality parameters such as oxidative stability and bitter index. The main differences were due to freeze injuries caused by low temperatures in December 2001. The levels of chlorophylls and carotenoids in olive oil or pulp from frost-damaged olive trees were lower as a consequence of faster ripening. The olive oil extracted from frost-damaged olive pulp had lower contents of secoiridoid and especially lower levels of 3,4-DHPEA-EDA (the dialdehydic form of elenolic acid linked to hydroxytyrosol). In the following crop seasons, a significant increase in phenolic compounds, especially in secoiridoid derivatives such as 3,4-DHPEA-EDA, was observed. This increase may be due to the fact that olive trees that suffered frost damage in December 2001 were more sensitive to stress caused by the water deficit during summer in the subsequent crop seasons, which is usual in this olive-growing region. Moreover, important correlation coefficients were observed between the main secoiridoid derivative compound (3,4-DHPEA-EDA) and oxidative stability and the bitter index.     

Effect of extraction systems on the phenolic composition of virgin olive oils

The effect of extraction systems on the phenolic composition of virgin olive oils obtained from two different Italian cultivars (Coratina and Oliarola) was determined. The oils extracted using two-phase centrifugation showed in all cases higher phenolic concentration in comparison to oils obtained from three-phase centrifugation. In particular, the highest differences were observed for aglykone derivatives of oleuropein (3,4-DHPEA-EDA and 3,4-DHPEA-EA) that are the most concentrated antioxidant phenolic compounds of virgin olive oil. These results were confirmed by the autoxidation stability of the oils examined.     

Impact of Aromatization by Citrus limetta and Citrus sinensis Peels on Olive Oil Quality,Chemical Composition and Heat Stability

The objectives of this work were to study the impact of aromatization by sweet lemon and sweet orange peels on olive oil quality, chemical composition and heat stability. Flavored olive oils were prepared by maceration of sweet lime and sweet orange zests at 1, 3 and 5 %. The oil samples were kept at 60 °C for 40 days. Physicochemical tests (FFA, PV, oxidative stability, polyphenols and pigments contents) were carried out. The antioxidant activity (β‐carotene‐linoleate bleaching assay, DPPH radical scavenging and reducing power) was also tested. Results indicated that the addition of citrus zests contributed to the increase in polyphenols and carotenoids contents. Moreover, it led to an increase in the DPPH‐radical scavenging activity (99.50 % for flavored olive oil with sweet orange zests at 5 %). Flavored olive oils with sweet orange zests were more stable to heat treatment than those flavored with sweet lemon zests. The degradation rate of bioactive compounds was lower for virgin olive oil comparing to flavored oils.     

Phenolic composition and antioxidant activity of Southern Italian monovarietal virgin olive oils

The phenolic composition and antioxidant activity of several monovarietal extra virgin olive oils used as blenders for the production of Collina di Brindisi protected designation of origin (PDO) oil, produced between December 2008 and January 2009 using two‐phases or three‐phases extraction system, were evaluated and compared with other manufacturer products designated as PDO. Oils were taken from the most representative ones industrial oil mills in the PDO geographical area. The parameters assessed were free acidity, peroxide value, K₂₃₂ and K₂₇₀ indices, organoleptic characteristics, total phenolic content (TPC), phenolic profile, and antioxidant activity coefficient (AAC). The phenolic contents and profiles of the monovarietal oils showed remarkable differences with respect to PDO oils. The variables that exerted a major influence on phenols concentration were the maturity degree of olives (December>January), followed by the extraction system (two‐phase>three‐phase), and place of growing. The Pearson r correlation index showed that AAC was positively correlated with TPC, p‐coumarate, and 3,4‐DHPEA‐EA, and negatively correlated with peroxide value. Practical applications: The results provide detailed information about: (i) the phenolic composition and the AAC of several monovarietal extra virgin olive oils used as blenders for the production of a PDO oil; (ii) the impact of genetic variability, place of growing, olive maturity degree, and extraction technology on oil phenol compounds; and (iii) the relationships among each phenolic compound and AAC, and their potential utilization as analytical index of antioxidant activity. It is important to study the phenolic compounds and antioxidant activity of monovarietal extra virgin olive oils used to produce PDO oil and to compare with the relative PDO samples in order to define a possible analytical tool able to verify what is stated in the label for consumer information and protection.     

Pinoresinol and 1-acetoxypinoresinol, two new phenolic compounds identified in olive oil

Polyphenols of olive oil show autoprotective, sensory, and nutritional-therapeutic effects. Two new phenolic compounds have been isolated from virgin olive oils by preparative high-performance liquid chromatography and their structures established on the basis of their mass spectra and nuclear magnetic resonance spectral data. The compounds identified are the lignans pinoresinol and 1-acetoxypinoresinol. Both have been found in all the commercial virgin olive oils analyzed. Pinoresinol concentration was rather similar in all the oils. In contrast, 1-acetoxypinoresinol concentration was higher in oils of the Arbequina and Empeltre cultivars than in Picual or Picudo cultivars. Pinoresinol and 1-acetoxypinoresinol may represent the major phenolic compounds in some Arbequina and Empeltre oils. Lignans possess biological and pharmacological properties and, therefore, the two new compounds identified in olive oils may contribute to the reported beneficial effects which are attributed to polyphenols on human health of a diet rich in olive oil.     

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.     

Potential of selected Portuguese cultivars for the production of high quality monovarietal virgin olive oil

The effect of cultivar and ripeness stage on the potential nutritional value of monovarietal extra virgin olive oils (MEVOOs) obtained from Cordovil, Carrasquinha, Verdeal, and Negrinha do Freixo cultivars was investigated. MEVOOs produced were characterized by high oleic acid (72–83%), tocopherol (182–530 mg/kg), and phenolic compounds (326–1110 mg/kg) content and by a similar polyphenolic profile. 1‐Penten‐3‐one was found to be the compound with the highest contribution for the aroma of the four MEVOO, related to bitter, pungent, and leaf attributes. MEVOO from Verdeal cultivar showed the best performance in terms of the composition: the highest yield of oil, the highest content of oleic acid, high tocopherol, polyphenol and sterol content, and the lowest content of linoleic acid. These characteristics give to these MEVOO not only a great oxidative stability but also interesting properties from the health point of view. MEVOO obtained with fruits at the maturity index of around 4 were in general richer in beneficial minor compounds. MEVOO produced were discriminated by variety and ripeness stage, using a stepwise linear discriminant analysis. This discrimination will in the future enable the prevention of adulteration of these monovarietal olive oils with specific nutritional composition with other olive oils. Practical implications: High‐quality MEVOOs have recently been introduced in the market, which for growers is a practical way to differentiate and increase the commercial value of extra virgin olive oil. The quantification of major and minor olive oil compounds in monovarietal olive oils represents an objective way of predicting the sensory characteristics, stability, and potential health benefits of the oils, as well as preventing their adulteration with other olive oils. This study will help in the selection of olive varieties during the maintenance or development of new olive orchards and also to select optimum harvest period for these varieties, in order to obtain MEVOOs with the maximum quality and health benefits for consumers.