Detection of the presence of refined hazelnut oil in refined olive oil by fluorescence spectroscopy

The fluorescence spectroscopy technique has been tested as regards its ability to differentiate between refined hazelnut and olive oils. Classification of these oils based on their excitation-emission fluorescence spectra data (spectral range 300-500 nm of the excitation spectra at lambdaem=655 and spectral range 650-900 of the emission spectra at lambdaex=50 nm) was performed using principal component analysis and artificial neural networks. Both methods provided good discrimination between the refined hazelnut and olive oils. The results have also pointed out the possibilities of a spectrofluorimetric method joined to multivariate analysis, to differentiate refined oils, and even to detect the presence of refined hazelnut oils in refined olive oils at percentages higher than 9%.     

Detection of hazelnut oil adulteration using FT-IR spectroscopy

Fourier transform infrared spectroscopy (FT-IR) was used to detect the adulteration of hazelnut oil with different types of oils and to detect the adulteration of extra-virgin olive oil with hazelnut oil. Spectra of hazelnut oil, seven other types of oils, extra-virgin olive oil, and the adulterated oils were collected with a FT-IR equipped with a ZnSe-ATR accessory and a MCTA detector. Discriminant analysis and partial least-squares analysis were used to analyze the data. Classification of hazelnut oil, olive oil, and the other types of oils was achieved successfully with FT-IR. The detection level for sunflower oil adulteration of hazelnut oil was 2%, and the correlation coefficient for the PLS model was 0.99. Adulteration of virgin olive oil with hazelnut oil could be detected only at levels of 25% and higher.     

Detection of extra virgin olive oil adulteration with lampante olive oil and refined olive oil using nuclear magnetic resonance spectroscopy and multivariate statistical analysis

High-field 31P NMR (202.2 MHz) spectroscopy was applied to the analysis of 59 samples from three grades of olive oils, 34 extra virgin olive oils from various regions of Greece, and from different olive varieties, namely, 13 samples of refined olive oils and 12 samples of lampante olive oils. Classification of the three grades of olive oils was achieved by two multivariate statistical methods applied to five variables, the latter being determined upon analysis of the respective 31P NMR spectra and selected on the basis of one-way ANOVA. The hierarchical clustering statistical procedure was able to classify in a satisfactory manner the three olive oil groups. Subsequent application of discriminant analysis to the five selected variables of oils allowed the grouping of 59 samples according to their quality with no error. Different artificial mixtures of extra virgin olive oil-refined olive oil and extra virgin olive oil-lampante olive oil were prepared and analyzed by 31P NMR spectroscopy. Subsequent discriminant analysis of the data allowed detection of extra virgin olive oil adulteration as low as 5% w/w for refined and lampante olive oils. Further application of the classification/prediction model allowed the estimation of the percent concentration of refined olive oil in six commercial blended olive oils composed of refined and virgin olive oils purchased from supermarkets.     

Rapid quantitative assessment of the adulteration of virgin olive oils with hazelnut oils using Raman spectroscopy and chemometrics

The authentication of extra virgin olive oil and its adulteration with lower-priced oils are serious problems in the olive oil industry. In addition to the obvious effect on producer profits, adulteration can also cause severe health and safety problems. A number of techniques, including chromatographic and spectroscopic methods, have recently been employed to assess the purity of olive oils. In this study Raman spectroscopy together with multivariate and evolutionary computational-based methods have been employed to assess the ability of Raman spectroscopy to discriminate between chemically very closely related oils. Additionally, the levels of hazelnut oils used to adulterate extra virgin olive oil were successfully quantified using partial least squares and genetic programming.     

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.     

Characterization of the total free radical scavenger capacity of vegetable oils and oil fractions using 2,2-diphenyl-1-picrylhydrazyl radical

The total free radical scavenger capacity (RSC) of 57 edible oils from different sources was studied: olive (24 brands of oils), sunflower (6), safflower (2), rapeseed (3), soybean (3), linseed (2), corn (3), hazelnut (2), walnut (2), sesame (2), almond (2), mixture of oils for salad (2), "dietetic" oil (2), and peanut (2). Olive oils were also studied according to their geographical origins (France, Greece, Italy, Morocco, Spain, and Turkey). RSC was determined spectrophotometrically by measuring the disappearance of the radical 2,2-diphenyl-1-picrylhydrazyl radical (DPPH(*)) at 515 nm. The disappearance of the radical followed a double-exponential equation in the presence of oils and oil fractions, which suggested the presence of two (fast and slow) groups of antioxidants. RSC was studied for the methanol-soluble phase ("methanolic fraction", MF) of the oil, the fraction nonsoluble in methanol ("lipidic fraction", LF), and the nonfractionated oil ("total oil"; TF = MF + LF). Only olive, linseed, rapeseed, safflower, sesame, and walnut oils showed significant RSC in the MF due to the presence of phenolic compounds. No significant differences were found in the RSC of olive oils from different geographical origins. Upon heating at 180 degrees C the apparent constant for the disappearance of RSC (k(T)) and the half-life (t1/2) of RSC for MF, LF, and TF were calculated. The second-order rate constants (k2) for the antiradical activity of some phenolic compounds present in oils are also reported.     

Effects of olive fruit quality and oil storage practices on the diacylglycerol content of virgin olive oils

The evolution of 1,3- and 1,2-isomers of diacylglycerols (DGs) in olive oils obtained from healthy olives and the influence of the olive quality was studied. Healthy olive fruits yielded oils containing almost exclusively 1,2-isomers whereas altered olives produced oils with significant amounts of 1,3-isomers. Virgin olive oils obtained from various olive cultivars and stored at different temperatures showed triacylglycerol hydrolysis and diacylglycerol isomerization depending on the acidity and temperature. The results indicated that the relationship between acidity and total diacylglycerol content has scarce utility for detecting mild refined oil in virgin olive oil. On the other hand, the 1,3-/1,2-DG isomers ratio is useful for assessing the genuineness of virgin olive oils with low acidities during the early stages of storage.     

Alkyl esters of fatty acids a useful tool to detect soft deodorized olive oils

Fatty acid alkyl esters (FAAEs) are a family of natural neutral lipids present in olive oils and formed by esterification of free fatty acids (FFAs) with low molecular alcohols. Inappropriate practices during the olive oil extraction process and bad quality of the olive fruits promote their formation. Quantification can be done by isolation with a silica gel solid phase extraction cartridge followed by analysis on a gas chromatograph equipped with a programmed temperature vaporizer injector using a polar capillary column. The application of the method to more than 100 Spanish olive oils from different categories, varieties, and geographical origin allowed for establishing the average content of FAAEs and distinguishing the Spanish protected denomination of origin (PDO) and extra virgin olive oils from other categories of olive oils. Those other categories of oils can be subjected to a mild refining process, which leads to blending with extra virgin olive oils. Studies on low quality oils subjected to mild refining showed that FAAEs remain after that process. Thereby, blends of extra virgin olive and mildly refined low quality olive oils can be detected by their alkyl ester concentrations.     

Performance of crude olive pomace oil and soybean oil during carotenoid production by Blakeslea trispora in submerged fermentation

Crude olive pomace oil (COPO) and crude soybean oil (CSO), two low-cost carbon sources, were examined as cosubstrates of glucose for carotenoid production by Blakeslea trispora. Results were compared to those obtained in glucose as a sole carbon source (medium 1) and glucose plus the respective end-line refined oil counterparts. Microbial growth in the presence of oils resulted in an increase in total carotenoid production. The performance of crude oils was better than that of the respective refined forms. Carotenoid production depended on both type and added oil amount. An increase in added oil amount did not necessarily favor carotenoid accumulation. The addition of 10 g oil/L of substrate stimulated carotenoid synthesis, mainly that of beta-carotene, more than 14 (COPO) and 40 times (CSO) in comparison to that observed in medium 1. The maximum total carotenoid content (as mg beta-carotene per g of biomass dry weight) was 75 (COPO) and 235 mg (CSO), respectively. Growth, substrate assimilation, and lipid accumulation-degradation also depended on the presence of oil in the substrate.     

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.     

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.     

Determination of proteins in refined and nonrefined oils

Five methods using aqueous/organic solvents for the separation of proteins from oils were compared. The extraction with acetone-hexane followed by amino acid analysis was found to be the most suitable method for isolation and quantification of proteins from oils. The detection limit of the method was 0.18 mg protein/kg oil, and the quantification limit was 0.6 mg protein/kg. The relative repeatability limit for samples containing 1-5 mg protein/kg sample was 27%. The protein recovery ranged between 68 and 133%. Using this method, the protein content of 14 refined and nonrefined oils was determined. In none of the refined oils were proteins detected, whereas the protein content of the unrefined oils ranged between undetectable in extra virgin olive oil to 11 mg/kg in rapeseed oil. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with silver staining, many protein bands were visible in the unrefined soy, olive, peanut, and rapeseed oil samples. Proteins bands were not obtained from the refined fish oil. In the other refined oil samples, a few proteins bands could be visualized. Two protein bands with apparent molecular molecular masses of 58 and 64 kDa were always observed in these oils.     

Interpretation of Fourier transform Raman spectra of the unsaponifiable matter in a selection of edible oils.

The unsaponifiable matter of edible oils is a source of information for their characterization and authentication. FT-Raman spectroscopy has been applied with success to the determination of the spectra of the unsaponifiable matter of varietal olive oils as well as other refined and crude edible oils. The spectra of the major unsaponifiable series of compounds (squalene, sterolic, and terpenic alcoholic fractions), together with beta-carotene and lutein, have been used to explain the most prominent bands found in the spectra of the unsaponifiable matter of 15 edible oil samples. The order of the scattering intensities of the varietal olive oils agrees with the results obtained by chromatography. An unsupervised multivariate statistical analysis of selected bands points out differences between olive oils and the other seed oils and also among varietal virgin olive oils.     

Polycyclic aromatic hydrocarbons and olive pomace oil

The occurrence of polycyclic aromatic hydrocarbons (PAHs) in five samples of olive pomace oil has been studied to determine the contamination degree of this type of oil and to evaluate if specific purification steps must be introduced during its manufacture. The PAHs present have been determined by gas chromatography-mass spectrometry. A high number of PAHs, with a wide range of molecular weights and in very high concentrations, have been found in four of the samples studied. A very high number of alkyl derivatives and, in many cases, in higher concentrations than their respective parent PAHs, have also been identified. One of the samples, however, presents a more reduced number of PAHs and in significantly lower concentrations than the others. These findings reveal that it is necessary to introduce adequate cleanup steps in the manufacturing process of olive pomace oil, which can give rise to oils with a relatively low content of PAHs. Some carcinogenic PAHs have also been identified, both unalkylated and alkylated.     

Determination of the diglyceride content in greek virgin olive oils and some commercial olive oils by employing (31)P NMR spectroscopy

In this study, the diglyceride contents of 96 samples of virgin olive oils from the regions of Crete, Lesvos, Messinia, Pilion, Zakynthos, Halkidiki, and Ilia, 15 samples of commercial extra virgin and pure olive oils, and 3 samples each of refined olive oils and pomace oils were determined by a facile method introduced in a previous publication. This method is based on the phosphitylation of the free hydroxyls of the diglycerides with 2-chloro-4,4,5,5-tetramethyldioxaphospholane and the integration of the appropriate peaks in the (31)P NMR spectra. This preliminary study showed interesting trends in the diglyceride content of the virgin olive oils from the various regions of Greece that can be used as simple criteria to assess the olive oil characteristics. Analysis of variance has been carried out for the diglyceride content of each region in an attempt to detect possible differences in the diglyceride levels among the various regions. Finally, the relationship between the ratio of 1,2-diglycerides to the total amount of diglycerides and the total amount of diglycerides has been used to monitor the quality of virgin olive oils, commercial olive oils, refined olive oils, and pomace oils.     

Authentication of vegetable oils by bulk and molecular carbon isotope analyses with emphasis on olive oil and pumpkin seed oil

The authenticity of vegetable oils consumed in Slovenia and Croatia was investigated by carbon isotope analysis of the individual fatty acids by the use of gas chromatography-combustion-isotope ratio mass spectrometry (GC/C/IRMS), and through carbon isotope analysis of the bulk oil. The fatty acids from samples of olive, pumpkin, sunflower, maize, rape, soybean, and sesame oils were separated by alkaline hydrolysis and derivatized to methyl esters for chemical characterization by capillary gas chromatography/mass spectrometry (GC/MS) prior to isotopic analysis. Enrichment in heavy carbon isotope ((13)C) of the bulk oil and of the individual fatty acids are related to (1) a thermally induced degradation during processing (deodorization, steam washing, or bleaching), (2) hydrolytic rancidity (lipolysis) and oxidative rancidity of the vegetable oils during storage, and (3) the potential blend with refined oil or other vegetable oils. The impurity or admixture of different oils may be assessed from the delta(13)C(16:0) vs. delta(13)C(18:1) covariations. The fatty acid compositions of Slovenian and Croatian olive oils are compared with those from the most important Mediterranean producer countries (Spain, Italy, Greece, and France).     

Multiclass pesticide determination in olives and their processing factors in olive oil: comparison of different olive oil extraction systems

The processing factors (pesticide concentration found in olive oil/pesticide concentration found in olives) of azinphos methyl, chlorpyrifos, lambda-cyhalothrin, deltamethrin, diazinon, dimethoate, endosulfan, and fenthion were determined in olive oil production process in various laboratory-scale olive oil extractions based on three- or two-phase centrifugation systems in comparison with samples collected during olive oil extractions in conventional olive mills located at different olive oil production areas in Greece. Pesticide analyses were performed using a multiresidue method developed in our laboratory for the determination of different insecticides and herbicides in olive oil by solid-phase extraction techniques coupled to gas chromatography detection (electron capture detection and nitrogen phosphorus detection), optimized, and validated for olive fruits sample preparation. Processing factors were found to vary among the different pesticides studied. Water addition in the oil extraction procedure (as in a three-phase centrifugation system) was found to decrease the processing factors of dimethoate, alpha-endosulfan, diazinon, and chlorpyrifos, whereas those of fenthion, azinphos methyl, beta-endosulfan, lambda-cyhalothrin, and deltamethrin residues were not affected. The water content of olives processed was found to proportionally affect pesticide processing factors. Fenthion sulfoxide and endosulfan sulfate were the major metabolites of fenthion and endosulfan, respectively, that were detected in laboratory-produced olive oils, but only the concentration of fenthion sulfoxide was found to increase with the increase of water addition in the olive oil extraction process.     

Identification of new steroidal hydrocarbons in refined oils and the role of hydroxy sterols as possible precursors

The dehydration of sterols during the refining process of vegetable oils results in the formation of steroidal hydrocarbons (sterenes or steradienes) with two double bonds in the ring system. Other steroidal hydrocarbons whose structures were in agreement with the presence of three double bonds in the ring system were detected in the sterene fractions of refined vegetable oils. The 5alpha-, 7alpha-, and 7beta-hydroxy derivatives of cholesterol and phytosterols have been dehydrated in n-butanol/H(3)PO(4) to form steroidal hydrocarbons with three double bonds at the 2, 4, and 6 positions in the ring system. These hydrocarbons had the same relative retention time and mass spectra as those present in the sterene fractions of refined oils. The dehydration of the hydroxy sterols dissolved in extra virgin olive oil and in the presence of 1% bleaching earths at 80 degrees C for 1 h results in the formation of the same steroidal hydrocarbons found in the refined oils.     

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.     

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.