Direct analysis of total antioxidant activity of olive oil and studies on the influence of heating

Aim of this study was to evaluate the total antioxidant activity (TAA) of extra virgin olive oil (EVOO) and the effect of heating on the alpha-tocopherol content and TAA in relation to the presence of polyphenols, heating time, and temperature. Experiments included the measurement by ABTS decolorization assay of antioxidant capacity of alpha-tocopherol and 14 simple phenolic compounds present in EVOO, either dissolved in ethanol or added to refined olive oil, and the evaluation of TAA, total phenols, and alpha-tocopherol of six commercial EVOO and three olive oils. Finally, four experimental oils were prepared from refined olive oil containing a fixed amount (300 ppm) of alpha-tocopherol and increasing amounts of polyphenols (25, 125, 225, and 326 ppm) extracted from EVOO. The thermal stability of experimental oils under domestic heating conditions (heating time from 30 to 120 min, heating temperature from 160 to 190 degrees C) was studied by evaluating the loss of alpha-tocopherol and TAA according to a Latin square design. Results indicate that TAA of commercial oils is mainly due to their phenol and alpha-tocopherol content. Heating experiments suggest that polyphenols from EVOO are effective stabilizers of alpha-tocopherol during olive oil heating, thus contributing to the nutritional value of cooked foods.     

Evolution of minor polar compounds and antioxidant capacity during storage of bottled extra virgin olive oil

We characterized "Olivastra Seggianese" extra virgin olive oil (EVOO) and evaluated its chemical and sensory characteristics and antioxidant and antiradical activities during storage under novel conditions. Two oils (A and B) were analyzed for the commodity characteristics at blending (t0) and after 9, 12, and 18 months; panel tests were performed and minor polar compounds (MPC) content was assessed at blending (t0) and after 6, 9, 12, and 18 months. Antioxidant and antiradical activities in vitro were evaluated at t0 and after 12 months, by human low density lipoprotein (LDL) and 1,1-diphenyl-2-picrylhydrazil radical (DPPH*) tests. Oil A, which had an initially higher MPC content, possessed "harder" organoleptic characteristics than oil B, which had a lower MPC content and was endowed with a "smoother" taste profile. Statistical analyses showed that secoiridoids, particularly deacetoxy-oleuropein aglycone, should be quantified to evaluate EVOO stability during storage. The antioxidant activity toward human LDL was linked to MPC content and to storage time. The tests on the stable free radical DPPH* confirmed the results on human LDL. We propose this as an additional parameter to evaluate olive oil quality and stability over time.     

Comparison of natural polyphenol antioxidants from extra virgin olive oil with synthetic antioxidants in tuna lipids during thermal oxidation

Polyphenols extracted from extra virgin olive oil (EVOO) were tested for their ability to inhibit lipid oxidation of canned tuna. Hydroperoxide formation during oxidation was monitored by measurement of peroxide value and decomposition of hydroperoxides by static headspace gas chromatographic analysis of volatiles. In tuna oxidized at 40 and 100 degrees C, 400 ppm of the EVOO polyphenols was an effective antioxidant as compared with 100 ppm of a 1:1 mixture of the synthetic antioxidants butylated hydroxytoluene and butylated hydroxyanisole. However, at concentrations <100 ppm, the EVOO phenolic compounds promoted hydroperoxide formation and decomposition. The EVOO polyphenols were effective antioxidants when added to heated tuna muscle in the presence of either brine or refined olive oil. The oxidation rate in tuna muscle packed in brine was higher than that of tuna packed in refined olive oil. The EVOO polyphenols had higher antioxidant activity in the brine samples than in the refined olive oil. The higher antioxidant activity of EVOO polyphenols in tuna packed in brine may be explained by their greater affinity toward the more polar interface between water and the fish oil system.     

Contribution of the phenolic fraction to the antioxidant activity and oxidative stability of olive oil

Antioxidant activity of the phenolic fraction of extra virgin olive oil (EVOO) was measured by means of a chemical and an electrochemical method. Both methods were tested in predicting the oxidative spoilage and stability to oxidation of 22 EVOO samples and resulted correlated with peroxide values and oxidative stability measured by Rancimat. The main phenolic compounds of EVOOs were detected by HRGC. To study the contribution of single polyphenols (PPs) to antioxidant activity of phenolic fraction and oxidative stability of EVOOs, multivariate statistical analyses were applied on HRGC data. An isomer of oleuropein aglycon was shown to affect significantly antioxidant activity of phenolic fraction but not oil stability to oxidation. No individual compounds was identified as the main cause of the overall antioxidant activity, and the total polyphenol determination by the Folin reagent was better correlated to antioxidant activity and oxidative stability than each tested PP or PPs groups such as o-diphenols.     

Excitation-emission fluorescence spectroscopy combined with three-way methods of analysis as a complementary technique for olive oil characterization

This paper shows the potential of excitation-emission fluorescence spectroscopy (EEFS) and three-way methods of analysis [parallel factor analysis (PARAFAC) and multiway partial least-squares (N-PLS) regression] as a complementary technique for olive oil characterization. The fluorescence excitation-emission matrices of a set of Spanish extra virgin, virgin, pure, and olive pomace oils were measured, and the relationship between them and some of the quality parameters of olive oils (peroxide value, K232, and K270) was studied. N-PLS was found to be more suitable than PARAFAC combined with multiple linear regression for correlating fluorescence and quality parameters, yielding better fits and lower prediction errors. The best results were obtained for predicting K270. EEFS allowed detection of extra virgin olive oils highly degraded at early stages (with high peroxide value) and little oxidized pure olive oils (with low K270). The proposed methodology may be used as an aid to analyze doubtful samples.     

Changes in virgin olive oil quality during low-temperature fruit storage

'Frantoio' olive fruits were stored at low temperature (4 +/- 2 degrees C) for 3 weeks to investigate the effect of postharvest fruit storage on virgin olive oil quality. Volatile compounds and phenolic compounds explained the changes in sensory quality that could not be explained with quality indices (FFA, PV, K232, and K270). Increases in concentrations of ( E)-2-hexenal and hexanal corresponded to positive sensory quality, whereas increases in ( E)-2-hexenol and (+)-acetoxypinoresinol were associated with negative sensory quality. Volatile and phenolic compounds were also indicative of the period of low-temperature fruit storage. Oleuropein and ligstroside derivatives in olive oil decreased with respect to storage time, and their significant ( p < 0.05) change corresponded to changes in bitterness and pungency. ( Z)-2-Penten-1-ol increased during low-temperature fruit storage, whereas 2-pentylfuran decreased. Changes in volatile compounds, phenolic compounds, quality indices, and sensory notes indicated that virgin olive oil quality was lost within the first week of low-temperature fruit storage and regained at 2 weeks. This research suggests that low-temperature olive fruit storage may be beneficial, with a possibility of increasing oil yield and moderating the sensory quality of virgin olive oils. This study demonstrates that deeper insights into virgin olive oil quality changes during low-temperature fruit storage may be gained by studying volatile and phenolic compounds in addition to quality indices and physical appearance of the fruit.     

New investigation of the isothermal oxidation of extra virgin olive oil: determination of free radicals, total polyphenols, total antioxidant capacity, and kinetic data

As a follow-up of the research programs carried out by our group concerning the artificial isothermal rancidification process in extra virgin olive oil (EVOO), in the present work the trends of both the total antioxidant capacity and the total polyphenols concentration as well as the main kinetic parameters of the process during the thermal oxidation of EVOO were studied and compared. In addition, the possibility of evaluating the increase in radicals concentration during the thermal oxidation process using a superoxide dismutase biosensor was also studied. The present investigation concerning this important food product is highly topical as it refers to the state of alteration of the EVOO used for cooking or frying, as a function of the temperature reached.     

Comparison of the radical scavenging potential of polar and lipidic fractions of olive oil and other vegetable oils under normal conditions and after thermal treatment

The antioxidant activity (IC(50)) of extra virgin olive oil (EVOO), commercial olive oil, and other vegetable oils (soybean, sunflower, and corn oil) was determined by UV-vis and by electron paramagnetic resonance (EPR) spectroscopy of the stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). Also, we studied the antioxidant activity of the methanol soluble phase (methanolic, MF) and the nonsoluble phase (lipidic, LF) of oils by the same methods. Similarly, we studied the effect of heating on the antioxidant activity at 160 and 190 degrees C. Also, the MF, containing the polyphenolic substances, was used for measurements of the radical scavenging capacity toward the most important oxygen free radicals, superoxide anion (O(2)(*)(-)) and hydroxyl (HO(*)) radicals. Results showed that soybean oil and EVOO had the highest antioxidant potential and thermal stability. In the case of soybean oil, the antioxidant capacity is the result of its high content of gamma- and delta-tocopherols (with the highest antioxidant capacity and thermostabilities), whereas in EVOO, the antioxidant potential is the result of the combination of specific antioxidant polyphenols, which are acting additionally as effective stabilizers of alpha-tocopherol. The high content of EVOO in tyrosol, hydrotyrosol, and oleuropein and other polyphenolics with radical scavenging abilities toward superoxide anion and hydroxyl radical suggests that olive oil possesses biological properties that could partially account for the observed beneficial health effects of the Mediterranean diet.     

Secoiridoids, tocopherols, and antioxidant activity of monovarietal extra virgin olive oils extracted from destoned fruits

The effect of olive stone removal before processing on the degradation level, secoiridoid and tocopherol contents, and antioxidant activity of monovarietal extra virgin olive oils (EVOOs) was studied. EVOOs were extracted from olives of the Leccino, Moraiolo, Frantoio, Pendolino, Taggiasca, and Colombaia varieties both in the presence and in the absence of the stones. The degradation level of EVOOs was evaluated by acidity, peroxide number, and spectroscopic indices K(232) and K(270), according to EU regulation. The secoiridoid compounds typical of EVOO, namely, the oleuropein and ligstroside derivatives, hydroxytyrosol, tyrosol, and tocopherols were analyzed by HPLC. The antioxidant activity was evaluated by the xanthine oxidase/xanthine system, generating superoxide radical and hydrogen peroxide, and by the 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl test. Results showed that EVOOs obtained from both stoned and destoned olives had a very low degradation level, which was not affected by destoning. Destoning lowered slightly the alpha-tocopherol content in EVOOs but increased the total secoiridoid content and the antioxidant activity of EVOOs (up to 3.5-fold). However, these effects were variety-dependent and negligible in some conditions. It was concluded that a better knowledge of the reactions occurring during olive processing, and particularly on the involvement of endogenous pulp and stone enzymes, is essential to predict the effect of destoning on EVOO quality.     

Changes in volatile and phenolic compounds with malaxation time and temperature during virgin olive oil production

Virgin olive oils produced at wide ranges of malaxation temperatures (15, 30, 45, and 60 degrees C) and times (30, 60, 90, and 120 min) in a complete factorial experimental design were discriminated with stepwise linear discriminant analysis (SLDA) revealing differences with processing conditions. Virgin olive oils produced at 15 and 60 degrees C for 30 min showed the most significant (p < 0.01) differences. Discrimination was based upon volatile and phenolic compounds detected in olive oils, peroxide value (PV), free fatty acids (FFA), ultraviolet (UV) absorbances, and oil yield. There were different discriminating variables for processing conditions illustrating the dependence of virgin olive oil quality on malaxation time and temperature. Volatile compounds were the dominant discriminating variables. Common oxidation indicators of olive oil (PV, K232, and K270) were not among the variables that significantly (p < 0.01) changed with malaxation time and temperature. Variables that discriminated both malaxation time and temperature were hexanal, 3,4-dihydroxyphenyl ethyl alcohol-decarboxymethyl elenolic acid dialdehyde (3,4-DHPEA-DEDA) and FFA, whereas 1-penten-3-ol, E-2-hexenal, octane, tyrosol, and vanillic acid significantly (p < 0.01) changed with temperature only and Z-2-penten-1-ol, (+)-acetoxypinoresinol, and oil yield changed with time only. Virgin olive oil quality was significantly influenced by malaxation temperature, whereas oil yield discriminated malaxation time. This study demonstrates the two modes of hexanal formation: enzymatic and nonenzymatic during virgin olive oil extraction.     

Discrimination of storage conditions and freshness in virgin olive oil

Virgin olive oil samples stored in the light at ambient temperature, in the dark at ambient temperature, and at low temperature in the dark for 12 months both with and without headspace were separated into recognizable patterns with stepwise linear discriminant analysis. The discrimination with variables volatile and phenolic compounds, free fatty acid (FFA), peroxide values, K232, and K270 revealed a departure of stored oil from freshness and showed significant (p < 0.01) differences between storage conditions. Virgin olive oil stored at low temperature had characteristics closest to fresh oil while oil stored in the light showed the largest departure from freshness. Parameters that exclusively and significantly (p < 0.01) discriminated storage conditions were identified as potential markers of the storage condition. In the presence of oxygen, hexanal was a marker of storage in the light, FFA was a marker for dark storage, and markers of low-temperature storage were acetic acid and pentanal. In the absence of oxygen, octane was the marker for storage in the light whereas tyrosol and hexanol were markers of virgin olive oil stored in the dark, with no marker indicative of low-temperature storage. E-2-Hexenal, K232, and K270 were identified as markers of virgin olive oil freshness.     

Influence of crystallization on the oxidative stability of extra virgin olive oil

The aim of this study was to evaluate the influence of the extra virgin olive oil (EVOO) physical state on the kinetics of oxidative reactions. To this purpose, EVOO was stored at increasing temperatures from 3 to 60 degrees C and the oxidation was followed by measuring both primary and secondary oxidation products. Results highlighted that crystallization plays an important role in determining EVOO stability. Below the melting point, the oxidation rate was found to be higher than that expected on the basis of the Arrhenius equation. The observed deviation from the Arrhenius equation was attributed to the physicochemical changes occurring as a consequence of phase transitions. In particular, the increase in unsaturated triacylglycerol concentration and the decrease of polyphenol content in the liquid phase surrounding fat crystals were indicated as the main factors causing the deviation. By taking into account these changes it was possible to describe the temperature dependence of the oxidation rate in the entire range of temperatures considered. This model appears to be promising in the challenge to find mathematical models able to predict the stability and, hence, the shelf life of lipid-containing foods.     

Nutritional and biological properties of extra virgin olive oil

The nutritional benefits generally recognized for the consumption of extra virgin olive oil (EVOO) are based on a large number of dietary trials of several international populations and intervention studies. Unfortunately, many authors in this field used questionable analytical methods and commercial kits that were not validated scientifically to evaluate the complex bioactive constituents of EVOO and lipid oxidation and decomposition products. Many questionable antiradical methods were commonly used to evaluate natural polyphenolic antioxidants, including an indirect method to determine low-density lipoprotein (LDL) cholesterol. Extensive differences were observed in experimental design, diet control, populations of different ages and problems of compliance intervention, and questionable biomarkers of oxidative stress. Analyses in many nutritional studies were limited by the use of one-dimensional methods to evaluate multifunctional complex bioactive compounds and plasma lipid profiles by the common applications of commercial kits. Although EVOO contains polyphenolic compounds that exhibit significant in vitro antioxidant activity, much more research is needed to understand the absorption and in vivo activity. Many claims of in vivo human beneficial effects by the consumption of EVOO may be overstated. No distinctions were apparently made between in vivo studies based on general health effects in large populations of human subjects and smaller scale well-controlled feeding trials using either pure or mixtures of known phenolic constituents of EVOO. More reliable protocols and testing methods are needed to better validate the complex nutritional properties of EVOO.     

Effects of enrichment of refined olive oil with phenolic compounds from olive leaves

The possibility of preparing olive oil, with the same nutritional value and stability characteristics found in virgin olive oil, by the enrichment of refined olive oil with olive leaf polyphenols was studied. To obtain antioxidant phenols similar to those found in virgin olive oil, these components were extracted from the leaves of several olive cultivars from the Northern region of Portugal, namely, Carrasca, Ripa, Negruche, Cordovil, Verdeal, Madural, and Bical cultivars, under several conditions. The concentration of a leaf extract required for addition to refined olive oil to obtain the same stability as virgin olive oil was determined. The extract from 1 kg of leaves was sufficient to fortify 50-320 L of refined olive oil to a similar stability as a virgin olive oil sample depending on the metal concentration of the oil, cultivar, and time of the year when the leaves were picked.     

Determinant parameters and components in the storage of virgin olive oil. Prediction of storage time beyond which the oil is no longer of "extra" quality.

This work studies the changes in the quality indices standardized by the European Union, together with the evolution of the oxidative stability and sterols, polyphenols, alpha-tocopherol, pigments, and fatty acids contents throughout the storage of Picual and Hojiblanca olive cultivar "extra" virgin olive oils at 2 degrees C + darkness and 30 degrees C + illumination. Only two quality indices (K(270) and sensory evaluation) indicate the loss of the extra quality of the oil during storage, and there is an excellent correlation between initial stability and the time to reach the limit of K(270) > 0.25 (after which the oil quality is no longer of "extra" quality). This time can be predicted with an error of <10%, which is of great commercial interest and previously unknown. Also unknown until now is that the changes in polyphenols, pigments, and alpha-tocopherol with storage time follow first-order kinetics.     

Confirmation of declared provenance of European extra virgin olive oil samples by NIR spectroscopy

The potential of near-infrared transflectance spectroscopy (1100-2498 nm) combined with chemometric techniques to confirm the geographical origin of European olive oil samples was evaluated. In total, 913 extra virgin olive oil samples (210 Ligurian and 703 non-Ligurian) were collected over three consecutive harvests (2005, 2006, and 2007). A multivariate spectral fingerprint for Ligurian olive oil was developed and deployed to confirm or refute a claim that any given sample was Ligurian. Samples were pseudorandomly split into calibration (n = 280) and validation sets (n = 633); the only selection constraint applied was to insist on equal numbers of Ligurian and non-Ligurian samples in the calibration set. Following preliminary examination by principal component analysis, the full spectrum modeling method applied to the spectral data set was discriminant partial least-squares regression; various data pretreatments were also investigated. The best models correctly predicted the origins of samples in the prediction set up to 92.8 and 81.5% for Ligurian and non-Ligurian olive oil samples, respectively, using a first-derivative data pretreatment. The potential of this approach in commercial traceability and quality assurance schemes is noted.     

Detection of rancid defect in virgin olive oil by the electronic nose

A sensor array of 32 conducting polymer sensors has been used to detect the rancid defect in virgin olive oils. A training set, composed of admixtures of a Portuguese virgin olive oil with different percentages (0-100%) of a rancid standard oil, was used for the selection of the best sensors classifying correctly the samples. Information on volatile compounds responsible for rancidity and the sensory evaluation of samples by assessors were used for explaining the mathematical selection of sensors. A tentative calibration, using unsupervised procedures (PCA and MDS) and a nonlinear regression, was carried out, with the training set, and later confirmed with a test set with which rancid commercial samples of different varieties were used to spike a Greek extra virgin olive oil at low levels of rancidity (0.5-6%).     

Antioxidant evaluation and oxidative stability of structured lipids from extravirgin olive oil and conjugated linoleic acid

Structured lipid (SL) was synthesized from extravirgin olive oil (EVOO) and conjugated linoleic acid (CLA) via a lipase-catalyzed reaction. CLA provides a variety of health benefits, but it is not consumed in free fatty acid form. The synthesized SL olive oil contained 42.5 mol % CLA isomers, and the major isomers were cis-9,trans-11-CLA (16.9 mol %) and trans-10,cis-12-CLA (24.2 mol %). The antioxidant activity determined by the radical scavenging capacity with the 2,2-diphenyl-1-picrylhydrazyl radical was lower in SL olive oil than in EVOO. The oxidative stability was also lower in SL olive oil since it had a higher peroxide value, rho-anisidine value, and 2-thiobarbituric acid reactive substances values during 20 days of storage at 60 degrees C. This observation could be due to the reduction in the natural phenolic compounds (97%) and tocopherols (56%), and the incorporated CLA with two conjugated double bonds in the SL olive oil. The oxidative stability of SL olive oil was increased by added rosemary extracts at concentrations of 100, 200, and 300 ppm. The present study suggests that the SL olive oil may be a suitable way to incorporate or deliver CLA into human diets. However, the addition of a proper antioxidant would be required for improving its oxidative stability.     

Phenolic compounds in olive oils intended for refining: formation of 4-ethylphenol during olive paste storage

The phenolic composition of "lampante olive oil", "crude olive pomace oil", and "second centrifugation olive oil" was characterized by high-performance liquid chromatography with UV, fluorescence, and mass spectrometry detection. The phenolic profile of these olive oils intended for refining was rather similar to that previously reported for virgin olive oil. However, a new compound was found in these oils, which is mainly responsible of their foul odor. It was identified as 4-ethylphenol by comparison of its UV and mass spectra with those of a commercial standard. Although 4-ethylphenol was discovered in all oils intended for refining, its presence was particularly significant in "second centrifugation olive oils", its concentration increasing with time of olive paste storage. Similar trends were observed for hydroxytyrosol, hydroxytyrosol acetate, tyrosol, and catechol, the concentration of these substances reaching values of up to 600 mg/kg of oil, which makes their recovery for food, cosmetic, or pharmaceutical purposes attractive.