Influence of simulated deep frying on the antioxidant fraction of vegetable oils after enrichment with extracts from olive oil pomace

The stability of the antioxidant fraction in edible vegetable oils has been evaluated during a simulated deep frying process at 180 °C. Four edible oils (i.e., extra-virgin olive oil with a 400 μg/mL overall content in naturally existing phenols; high-oleic sunflower oil without natural content of these compounds but enriched either with hydrophilic antioxidants isolated from olive pomace or with an oxidation inhibitor, dimethylsiloxane; and sunflower oil without enrichment) were subjected to deep heating consisting of 20 cycles at 180 °C for 5 min each. An oil aliquot was sampled after each heating cycle to study the influence of heating on the antioxidant fraction composed of hydrophilic and lipophilic antioxidants such as phenols and tocopherols, respectively. The decomposition curves for each group of compounds caused by the influence of deep heating were studied to compare their resistance to oxidation. Thus, the suitability of olive pomace as raw material to obtain these compounds offers an excellent alternative to the use of olive-tree materials different from leaves. The enrichment of refined edible oils with natural antioxidants from olive pomace is a sustainable strategy to take benefits from this residue.     

Liquid-liquid extraction for the enrichment of edible oils with phenols from olive leaf extracts

A liquid-liquid extraction method to enrich edible oils--olive, sunflower, and soy oils--with phenols from olive leaf extracts is proposed. After microwave assistance to remove the phenols from three varieties of olive leaves, concentrations in the extracts between 12921 and 5173 mg/L of oleuropein, between 488 and 192 mg/L of apigenin-7-glucoside, between 444 and 219 mg/L of luteolin-7-glucoside, and between 501 and 213 mg/L of verbascoside were obtained, which clearly depended on the target variety. After optimization of the liquid-liquid extraction step, the concentrations in oils were 442, 162, and 164 mg/L of oleuropein, respectively, which were also enriched in apigenin-7-glucoside (between 8 and 15 mg/L, depending of the oil), lutelin-7-glucoside (between 11 and 12 mg/L), and verbascoside (between 11 and 13 mg/L). The oil-extract distribution factor of these compounds was also calculated for all olive leaf varieties and edible oils using different extracts concentrations and also different oil-extract volume ratios. Thus, a door is open to enrichment of any oil with olive phenols at preset concentrations using extracts preconcentrated as required and taking into account the distribution factor of the target compounds between the oil and the extracts.     

Solid-liquid transfer of biophenols from olive leaves for the enrichment of edible oils by a dynamic ultrasound-assisted approach

A continuous approach assisted by ultrasound for direct enrichment of edible oils (olive, sunflower, and soya) with the main phenols in olive leaves (i.e., oleuropein, verbascoside, apigenin-7-glucoside, and luteolin-7-glucoside) has been developed. Multivariate methodology was used to carry out a detailed optimization of the enrichment, and quantitation of the transferred compounds was based on LC-MS-MS in multiple reaction monitoring optimizing the most sensitive transition for each biophenol. Under the optimal working conditions, only 20 min is necessary to enrich the edible oils with 14.45-9.92 microg/mL oleuropein, 2.29-2.12 microg/mL verbascoside, 1.91-1.51 microg/mL apigenin-7-glucoside, and 1.60-1.42 microg/mL luteolin-7-glucoside. The enrichment method is carried out at room temperature and is organic-solvent-free; thus, the healthy properties of the edible oils improve as does their quality. Also, the low acquisition and maintenance costs of an ultrasound source and its application in a dynamic system make advisable the industrial implementation of the proposed method.     

Development of a phenol-enriched olive oil with both its own phenolic compounds and complementary phenols from thyme

Besides affecting the oil's sensorial characteristics, the presence of herbs and spices has an impact on the nutritional value of the flavored oils. The aim of the study was to develop a new product based on the phenol-enrichment of a virgin olive oil with both its own phenolic compounds (secoiridoid derivatives) plus additional complementary phenols from thyme (flavonoids). We studied the effect of the addition of phenolic extracts (olive cake and thyme) on phenolic composition, oxidative stability, antioxidant activity, and bitter sensory attribute of olive oils. Results showed that flavonoids from thyme appeared to have higher transference ratios (average 89.7%) from the phenolic extract to oil, whereas secoiridoids from olive presented lower transference ratios (average 35.3%). The bitter sensory attribute of the phenol-enriched oils diminished with an increase of the concentration of phenols from thyme, which might denote an improvement in the consumer acceptance.     

Olea europaea L. leaf extract and derivatives: antioxidant properties

This paper reports a very simple and fast method to collect eluates with high amounts of hydroxytyrosol, biotransforming Olea europaea L. leaf extract by a thermophilic beta-glycosidase immobilized on chitosan. Some phenolic compounds in the leaf tissue and in the eluates obtained by biotransformation are identified. To propose the eluates as natural substances from a vegetal source, their antioxidant properties have been compared with those of the leaf extract from which they are originated. The eluates possess a higher concentration of simple phenols, characterized by a stronger antioxidant capacity, than those available in extra virgin olive oils and in many tablets of olive leaf extracts, commercially found as dietetic products and food integrators.     

Influence of deep frying on the unsaponifiable fraction of vegetable edible oils enriched with natural antioxidants

The influence of deep frying, mimicked by 20 heating cycles at 180 °C (each cycle from ambient temperature to 180 °C maintained for 5 min), on the unsaponifiable fraction of vegetable edible oils represented by three characteristic families of compounds (namely, phytosterols, aliphatic alcohols, and triterpenic compounds) has been studied. The target oils were extra virgin olive oil (with intrinsic content of phenolic antioxidants), refined sunflower oil enriched with antioxidant phenolic compounds isolated from olive pomace, refined sunflower oil enriched with an autoxidation inhibitor (dimethylpolysiloxane), and refined sunflower oil without enrichment. Monitoring of the target analytes as a function of both heating cycle and the presence of natural antioxidants was also evaluated by comparison of the profiles after each heating cycle. Identification and quantitation of the target compounds were performed by gas cromatography-mass spectrometry in single ion monitoring mode. Analysis of the heated oils revealed that the addition of natural antioxidants could be an excellent strategy to decrease degradation of lipidic components of the unsaponifiable fraction with the consequent improvement of stability.     

Differentiation of refined and virgin edible oils by means of the trans- and cis-phytol isomer distribution

The differentiation of nonrefined (e.g., cold-pressed) and refined edible oils is an important task in food control because of the higher commercial value of the former. Here, we explored the suitability of the relative abundance of cis-phytol as a marker for authentication of nonrefined edible oils. Phytol, the tetramethyl-branched, monoenoic alcohol, is found widespread in nature as a part of chlorophyll. In chlorophyll, only trans-phytol is found. In this study, we present a method for the analysis of the phytol isomers, considering that traces of cis-phytol (contributing 0.1% to the phytol content) can be determined next to trans-phytol. For this purpose, phytol was gathered with the unsaponifiable matter from the oil, trimethylsilylated, and analyzed by gas chromatography coupled to mass spectrometry. With this method, 27 samples of edible oils (16 refined and 11 nonrefined edible oils) were analyzed for the abundance of cis-phytol relative to trans-phytol. In the nonrefined oils (e.g., olive oil, rapeseed oil, maize oil, and sunflower oil), cis-phytol contributed 0.1% (n = 3) or less (n = 8) to the phytol content. In contrast, the refined olive oils (n = 4) contained a share of 1.3-3% cis-phytol; the refined rapeseed oil (n = 3) contained a share of 0.7-1.0% cis-phytol; and the refined sunflower oil (n = 4) contained a share of 0.3-0.9% cis-phytol. Only one refined pomegranate kernel did not contain cis-phytol. The phytol concentration was not suited to distinguish nonrefined from refined oils. In contrast, our data suggest that the virtual absence of cis-phytol can be used as a marker for nonrefined (e.g., cold-pressed) edible oils.     

Qualitative and semiquantitative analysis of phenolic compounds in extra virgin olive oils as a function of the ripening degree of olive fruits by different analytical techniques

Capillary electrophoresis (CE) can be effectively used as a fast screening tool to obtain qualitative and semiquantitative information about simple and complex phenolic compounds of extra virgin olive oil. Three simple phenols (tyrosol, hydroxytyrosol, and vanillic acid), a secoiridoid derivative (deacetoxy oleuropein aglycon), and two lignans (pinoresinol and acetoxypinoresinol) were detected as the main compounds in extra virgin olive oils by high-performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE). Spectrophotometric indices, radical scavenging activity, and oxidative stability of extra virgin olive oil samples obtained from olives hand-picked at different ripening degrees were statistically correlated with the CZE and HPLC quantification. The concentration of phenols in extra virgin olive oil decreased with ripeness of olive fruits. The high correlations found between CZE and the other analytical results indicate that CE can be applied as a rapid and reliable tool to routinely determine phenolic compounds in extra virgin olive oils.     

Supplementation of plasma with olive oil phenols and extracts: influence on LDL oxidation

Phenols present in olive oil may contribute to the health effects of the Mediterranean lifestyle. Olive oil antioxidants increase the resistance of low-density lipoproteins (LDL) against oxidation in vitro, but human intervention studies have failed to demonstrate similar consistent effects. To better mimic the in vivo situation, plasma was incubated with either individual olive oil phenols or olive oil extracts with different phenolic compositions, and LDL was subsequently isolated and challenged for its resistance to oxidation. The results show that the ortho-dihydroxy phenols (hydroxytyrosol and oleuropein-aglycone) are more efficient than their mono-hydroxy counterparts (tyrosol and ligstroside-aglycone) in increasing the resistance of LDL to oxidation. However, the concentration of antioxidants required to inhibit LDL oxidation when added to whole plasma was substantially higher as compared to previous data where antioxidants are directly added to isolated LDL. In conclusion, this study supports the hypothesis that extra virgin olive oil phenols protect LDL in plasma against oxidation. The explanation that in vitro studies show protective effects in contrast to the lack of effect in the majority of human studies may be that the dose of the phenols and thus their plasma concentration in humans was too low to influence ex vivo LDL oxidizability. Further studies are required to gain a better understanding of the potential health benefits that extra virgin olive oil may provide.     

Cluster analysis applied to the exploratory analysis of commercial spanish olive oils by means of excitation-emission fluorescence spectroscopy

Olive oil fluorescence is related to oil composition. Here it is shown that the natural clustering of different types of commercial Spanish olive oils depends on their fluorescence excitation-emission matrices (EEMs). Fifty-six commercial samples of olive oil (29 virgin olive oils, 20 pure olive oils, and 7 olive-pomace oils) were used. The clustering method was hierarchical agglomerative clustering using the Euclidean distance as a similarity measure and the average linkage. Two spectral ranges were considered (which either contained the fluorescence peak of the chlorophylls or did not), and various methods for preprocessing the fluorescence spectra were compared. The oils were clearly distinguished using the unfolded EEMs measured between lambda(ex) = 300-400 nm and lambda(em) = 400-600 nm. The optimal preprocessing was normalization of the unfolded spectra followed by column autoscaling. Also shown are the advantages of using second-order data (EEMs) instead of first-order data (a single fluorescence spectrum) for each sample.     

Evaluation of the influence of thermal oxidation on the phenolic composition and on the antioxidant activity of extra-virgin olive oils

A comparison between the results obtained by using HPLC-UV, HPLC-MS, and CE-UV for characterizing the deterioration of extra-virgin olive oil during heating (180 degrees C) was investigated, taking into account phenolic compounds. The concentration of several compounds belonging to four families of phenols (simple phenols, lignans, complex phenols, and phenolic acids) was determined in the samples after the thermal treatment by all three techniques. Hydroxytyrosol, elenolic acid, decarboxymethyl oleuropein aglycon, and oleuropein aglycon reduced their concentration with the thermal treatment more quickly than other phenolic compounds present in olive oil. HYTY-Ac and Lig Agl were demonstrated to be quite resistant to this kind of treatment, and the behavior of lignans could be outstanding, as they belong to the family most resistant to thermal treatment. Several "unknown" compounds were determined in the phenolic profiles of the oils after the thermal treatment, and their presence was confirmed in refined olive oils. The oxidative stability index (OSI time) was reduced from 25 to 5 h after 3 h of heating, whereas the peroxide value showed a minimum after 1 h of heating.     

Effects of olive, canola, and sunflower oils on the formation of volatiles from the Maillard reaction of lysine with xylose and glucose

Some important edible oils (extra virgin olive oil, canola oil, and sunflower oil) were added to aqueous glucose-lysine or xylose-lysine model systems to investigate their effect on the formation of volatiles from the Maillard reaction (MR). The volatile compounds were extracted by a Likens-Nickerson apparatus and quantified. Pyrazines, Maillard reaction products with an important impact on food flavor, appeared to be particularly sensitive to the presence of the oils in both the xylose-lysine and glucose-lysine model systems. The unsubstituted pyrazine was formed more with olive oil, less with canola oil, and even less with sunflower oil, whereas 2-methylpyrazine, 2,5-methylpyrazine, and 2,3-dimethylpyrazine were formed less with olive oil, more with canola oil, and even more with sunflower. The oxidative states of the oils and their fatty acid fingerprints were determined: the results indicated that the relative amounts of the pyrazines are sensitive to the degree of unsaturation of the oil. The autoxidation of the volatile compounds generated from the MR, investigated by the addition of free radical modulators (antioxidants alpha-tocopherol, 2,6-di-tert-butyl-4-methylphenol, and rosemary extract; or pro-oxidant alpha,alpha'-azobis-isobutyronitrile, a free radical initiator), was limited in respect to aqueous model systems.     

Oil stability prediction by high-resolution (13)C nuclear magnetic resonance spectroscopy

(13)C NMR spectra of oil fractions obtained chromatographically from 66 vegetable oils were obtained and analyzed to evaluate the potential use of those fractions in predicting oil stabilities and to compare those results with oil stability prediction by using chemical determinations. The oils included the following: virgin olive oils from different cultivars and regions of Europe and north Africa; "lampante" olive, refined olive, refined olive pomace, low-erucic rapeseed, high-oleic sunflower, corn, grapeseed, soybean, and sunflower oils. Oils were analyzed for fatty acid and triacylglycerol composition, as well as for phenol and tocopherol contents. By using stepwise linear regression analysis (SLRA), the chemical determinations and the (13)C NMR data that better explained the oil stability determined by the Rancimat were selected. These selected variables were related to both the susceptibility of the oil to be oxidized and the content of minor components that most contributed to oil stability. Because (13)C NMR considered many more variables than those determined by chemical analysis, the predicted stabilities calculated by using NMR data were always better than those obtained by using chemical determinations. All these results suggest that (13)C NMR may be a powerful tool to predict oil stabilities when applied to chromatographically enriched oil fractions.     

Detecting and quantifying sunflower oil adulteration in extra virgin olive oils from the eastern mediterranean by visible and near-infrared spectroscopy

One hundred and thirty-eight oil samples have been analyzed by visible and near-infrared transflectance spectroscopy. These comprised 46 pure extra virgin olive oils and the same oils adulterated with 1% (w/w) and 5% (w/w) sunflower oil. A number of multivariate mathematical approaches were investigated to detect and quantify the sunflower oil adulterant. These included hierarchical cluster analysis, soft independent modeling of class analogy (SIMCA method), and partial least squares regression (PLS). A number of wavelength ranges and data pretreatments were explored. The accuracy of these mathematical models was compared, and the most successful models were identified. Complete classification accuracy was achieved using 1st derivative spectral data in the 400-2498 nm range. Prediction of adulterant content was possible with a standard error equal to 0.8% using 1st derivative data between 1100 and 2498 nm. Spectral features and chemical literature were studied to isolate the structural basis for these models.     

Enzymatic assay for the determination of olive oil polyphenol content: assay conditions and validation of the method

A new spectrophotometric assay for the determination of the polyphenolic content of olive oil is presented. It is a substrate-recycling assay for phenolic compounds that employs tyrosinase in the presence of excess NADH. The reaction of various phenols with the enzyme produces an o-quinone, which is detected by recycling between reactions with the enzyme and NADH. The method offers some advantages over the classical methods employed to determine the polyphenolic content of olive oil, that is, ease and reproducibility of the analysis, highly increased sensitivity, and selectivity toward phenolic compounds. The amount of total polyphenols was determined in virgin olive oils both with the Folin-Ciocalteu reagent and with the proposed enzymatic method. The results suggest a better estimation of the polyphenol content, as compared with the colorimetric method. This has to be attributed to the different reactivities of the two methods toward phenols and catechols. Finally, the enzymatic method demonstrates that there is a linear relationship between the olive oil phenolic content and the antioxidative capacity of oil extracts.     

Phenolic compounds in Spanish olive oils.

Phenolic compounds in Spanish virgin olive oils were characterized by HPLC. Simple phenols such as hydroxytyrosol, tyrosol, vanillic acid, p-coumaric acid, ferulic acid, and vanillin were found in most of the oils. The flavonoids apigenin and luteolin were also found in most of the oils. The dialdehydic form of elenolic acid linked to tyrosol and hydroxytyrosol was also detected, as were oleuropein and ligstroside aglycons. The structure of a new compound was elucidated by MS and NMR as being that of 4-(acetoxyethyl)-1,2-dihydroxybenzene. Changes of phenolic compounds in virgin olive oils with maturation of fruits were also studied. Hydroxytyrosol, tyrosol, and luteolin increased their concentration in oils with maturation of fruits. On the contrary, glucoside aglycons diminished their concentration with maturation. No clear tendency was observed for the rest of the phenolic compounds identified.     

Determination of phenols, flavones, and lignans in virgin olive oils by solid-phase extraction and high-performance liquid chromatography with diode array ultraviolet detection

A simple analytical method for the quantitative determination of phenols, flavones, and lignans in virgin olive oils was developed. The polar fraction was isolated from small amounts of oil sample (2.5 g) by solid-phase extraction (SPE) using diol-phase cartridges, and the extract was analyzed by reversed-phase HPLC coupled with diode array UV detection. Chromatographic separation of pinoresinol, cinnamic acid, and 1-acetoxypinoresinol was achieved. Repeatability (RSD < 6.5%), recovery (> 90%), and response factors for each identified component were determined. SPE on amino-phase cartridges was used for isolating acidic phenols and as an aid for phenol identification. For the first time, 2-(4-hydroxyphenyl)ethyl acetate was detected in olive oils. The aldehydic structure of the ligstroside aglycon was confirmed by NMR spectroscopy. The colorimetric determination of total o-diphenolic compounds by reaction with molybdate was consistent with their HPLC determination. Differences between results obtained by liquid-liquid extraction and SPE were not statistically significant.     

Formation and evolution of monoepoxy fatty acids in thermoxidized olive and sunflower oils and quantitation in used frying oils from restaurants and fried-food outlets

The formation and evolution of monoepoxy fatty acids, arising from oleic and linoleic acids, were investigated in olive oil and conventional sunflower oil, representatives of monounsaturated and polyunsaturated oils, respectively, during thermoxidation at 180 degrees C for 5, 10, and 15 h. Six monoepoxy fatty acids, cis-9,10- and trans-9,10-epoxystearate, arising from oleic acid, and cis-9,10-, trans-9,10-, cis-12,13-, and trans-12,13-epoxyoleate, arising from linoleic acid, were analyzed by gas chromatography after oil derivatization to fatty acid methyl esters. Considerable amounts, ranging from 4.29 to 14.24 mg/g of oil in olive oil and from 5.10 to 9.44 mg/g of oil in sunflower oil, were found after the heating periods assayed. Results showed that the monoepoxides quantitated constituted a major group among the oxidized fatty acid monomers formed at high temperature. For similar levels of degradation, higher contents of the monoepoxides were found in olive oil than in sunflower oil. Ten used frying oils from restaurants and fried-food outlets in Spain were analyzed to determine the contents of the monoepoxides in real frying oil samples. Levels ranged from 3.37 to 14.42 mg/g of oil. Results show that, for similar degradation levels, the monoepoxides were more abundant in the monounsaturated oils than in the polyunsaturated oils.     

Antithrombotic lipid minor constituents from vegetable oils. Comparison between olive oils and others

Many epidemiological studies suggest that vegetable oils and especially olive oil present a protective effect against atherosclerosis. In this study, total lipids (TL) of Greek olive oils and seed oils of four kinds, namely, soybean, corn, sunflower, and sesame oil, were separated into total polar lipids (TPL) and total neutral lipids (TNL) via a novel extraction procedure. TPL and TNL of olive oil were fractionated by HPLC for further study. Each lipid fraction from HPLC separation along with TL, TPL, and TNL lipid samples from oils were tested in vitro for their capacity to induce or to inhibit washed rabbit platelet aggregation. Comparison between olive and seed oils supports the superiority of olive oil as high levels of platelet activating factor (PAF) antagonists have been detected, mainly in TPL. In addition, the structure of the most active fraction from olive oil was elucidated, as a glycerol-glycolipid. Because it has already been reported that PAF plays a pivotal role in atherogenesis, the existence of PAF agonists and antagonists in vegetable oils may explain their protective role against atherosclerosis.     

Comparative study of virgin olive oil quality from single varieties cultivated in Chile and Spain

Olive tree varieties that were cultivated only in the Mediterranean basin a few decades ago are now planted in the Southern Hemisphere as well. The chemical composition of the oils produced in countries as far distant as Spain and Chile are affected by differences in latitude and climate. In this work, seven monovarietal virgin olive oils from Chile (Arbequina, Barnea, Frantoio, Koroneiki, Leccino, Manzanilla and Picual) have been characterized by the chemical compounds responsible for taste (phenols) and aroma (volatiles). The oils were produced in five regions of Chile, and the concentration values of some chemical compounds were related to the geographical location of the olive tree orchards. Virgin olive oils from the major cultivars, Arbequina and Picual, were characterized in comparison with the same monovarietal oils produced in Spain. The concentration values of fourteen volatile compounds showed significant differences (p < 0.05) between the oils produced in Spain and Chile. Concerning the phenol composition, main differences were found on the secoiridoids derivatives of oleuropein and ligstroside, apigenin and luteolin.