Health and sensory properties of virgin olive oil hydrophilic phenols: agronomic and technological aspects of production that affect their occurrence in the oil

Hydrophilic phenols are the most abundant natural antioxidants of virgin olive oil (VOO), in which, however, tocopherols and carotenes are also present. The prevalent classes of hydrophilic phenols found in VOO are phenolic alcohols, phenolic acids, flavonoids, lignans and secoiridoids. Secoiridoids, that include aglycon derivatives of oleuropein, demethyloleuropein and ligstroside, that are present in olive fruit, are the most abundant phenolic antioxidants of VOO. The sensory and healthy proprieties of VOO hydrophilic phenols as well as the agronomic and technological parameters that affect their concentration in the oil are discussed in this paper.     

Comparative elucidation of phenolic compounds in Albanian olive oils using LC-DAD-ESI-MS/MS

Abstract

Olive oils may provide health benefits, including the prevention of coronary heart diseases, cancers, and the modification of immune and inflammatory responses. These benefits mainly originate from the phenolic compounds found in olive oil. There has been no study on the advanced characterization of Albanian olive oils from various cultivars regarding phenolic compounds. Hence, a comprehensive characterization of phenolic compounds is carried out in Albanian monocultivar virgin olive oils from five different cultivars, including Kalinjot, Bardhi Tirana, Ulliri-i-Zi Tirana, Krips Kruja, and Bardhi Kruja for the first time. Liquid chromatography coupled to diode array detection and electrospray ?onization tandem mass spectrometry (LC-DAD-ESI-MS/MS) is employed for the determination of phenolic compounds. In total, 18 compounds were identified in all samples, including phenolic alcohols, phenolic acids, secoiridoids, flavonoids, and phenolic aldehydes. Significant quantitative differences were detected among the cultivars, with the highest concentrations detected in virgin olive oil (VOO) from cv. Ulli-i-Zi. Secoiridoids were found in abundance, in general, followed by phenolic alcohols, and in this group, 3,4-DHPEA-EDA and p-HPEA-EDA stood out as dominant compounds, especially in Kalinjot virgin olive oils. Regarding phenolic alcohols, 3,4-DHPEA-AC was determined as the main phenolic compound. Phenolic profiles were found to be significantly different among the olive oil samples of different cultivars. Principal component analyses (PCA) displayed the differentiation of samples in terms of phenolic compounds.     

Analytical determination of polyphenols in olive oils

The increasing popularity of olive oil is mainly attributed to its high content of oleic acid, which may affect the plasma lipid/lipoprotein profiles, and its richness in phenolic compounds, which act as natural antioxidants and may contribute to the prevention of human disease. An overview of analytical methods for the measurement of polyphenols in olive oil is presented. In principle, the analytical procedure for the determination of individual phenolic compounds in virgin olive oil involves three basic steps: extraction from the oil sample, analytical separation, and quantification. A great number of procedures for the isolation of the polar phenolic fraction of virgin olive oil, utilizing two basic extraction techniques, LLE or SPE, have been included. The reviewed techniques are those based on spectrophotometric methods, as well as analytical separation (gas chromatography (GC), high-performance liquid chromatography (HPLC), and capillary electrophoresis (CE)). Many reports in the literature determine the total amount of phenolic compounds in olive oils by spectrophometric analysis and characterize their phenolic patterns by capillary gas chromatography (CGC) and, mainly, by reverse phase high-performance liquid chromatography (RP-HPLC); however, CE has recently been applied to the analysis of phenolic compound of olive oil and has opened up great expectations, especially because of the higher resolution, reduced sample volume, and analysis duration. CE might represent a good compromise between analysis time and satisfactory characterization for some classes of phenolic compounds of virgin olive oils.     

Retention effects of oxidized polyphenols during analytical extraction of phenolic compounds of virgin olive oil

The hydrophilic extract of virgin olive oil contains several phenolic compounds such as simple phenols, lignans, and secoiridoids that have been widely studied in recent years. Interest in the hydrophilic extract has also been extended to the fraction of oxidized phenols that form during storage as a consequence of oxidative stress. The present investigation compares the two most commonly used extraction methods, namely liquid-liquid extraction and SPE, on fresh virgin olive oil and that kept at different temperatures in the presence of oxygen to promote the formation of oxidative products. The selective retention of these natural and oxidized phenolic compounds in relation to the extraction method was assessed. Quantification of eight identified phenolic molecules and 11 unknown peaks was performed by HPLC-DAD/MSD.     

Study of the phenolic composition of spanish and italian monocultivar extra virgin olive oils: Distribution of lignans, secoiridoidic, simple phenols and flavonoids

The aim of the present study was to compare the phenolic content in selected monocultivar extra virgin olive oils. Analyses were carried out by HPLC/DAD/MS on Picual, Picuda, Arbequina and Hojiblanca oils from Spain and Seggianese and Taggiasca oils from Italy. Oils from cultivar Picual showed similar characteristics to those of Seggianese oils, with total amounts of secoiridoids of 498.7 and 619.2 mg/L, respectively. The phenolic composition of Arbequina oils is close to that of the Taggiasca variety with lignans among the main compounds. The determination of free and linked OH-Tyr, by way of an acid hydrolysis, represents a rapid and suitable method, especially when standards are not available, to determine antioxidant potentialities in terms of MPC, particularly for fresh extra virgin olive oils rich in secoiridoidic derivatives.     

Applicability of an In-Vitro Digestion Model to Assess the Bioaccessibility of Phenolic Compounds from Olive-Related Products

The Mediterranean diet includes virgin olive oil (VOO) as the main fat and olives as snacks. In addition to providing nutritional and organoleptic properties, VOO and the fruits (olives) contain an extensive number of bioactive compounds, mainly phenolic compounds, which are considered to be powerful antioxidants. Furthermore, olive byproducts, such as olive leaves, olive pomace, and olive mill wastewater, considered also as rich sources of phenolic compounds, are now valorized due to being mainly applied in the pharmaceutical and nutraceutical industries. The digestive system must physically and chemically break down these ingested olive-related products to release their phenolic compounds, which will be further metabolized to be used by the human organism. The first purpose of this review is to provide an overview of the current status of in-vitro static digestion models for olive-related products. In this sense, the in-vitro gastrointestinal digestion methods are widely used with the following aims: (i) to study how phenolic compounds are released from their matrices and to identify structural changes of phenolic compounds after the digestion of olive fruits and oils and (ii) to support the functional value of olive leaves and byproducts generated in the olive industry by assessing their health properties before and after the gastrointestinal process. The second purpose of this review is to survey and discuss all the results available to date.     

Effect of storage on quality parameters and phenolic content of Italian extra-virgin olive oils

Abstract

The quality of extra virgin olive oils is affected mainly by hydrolytic and oxidative reactions. The present paper investigated the changes of major and minor components and oxidation indices of three monovarietal extra virgin olive oils after 18?months of storage at room temperature and in dark glass bottles conditions. After storage, the basic quality parameters such as free acidity, peroxide values, extinction coefficients, fatty acids composition, chlorophyll and carotenoid content, did not exceed the upper limits set by European Community Regulations for extra-virgin olive oils. Given the importance of the phenolic fraction, UHPLC-HESI-MS metodology was used. A decrease in 3,4-DHPEA-EDA (oleacin) and p-HPEA-EDA (oleochantal) was detected whereas, an increase of tyrosol and hydroxytyrosol was measured as a consequence of degradation of ligstroside and oleuropein derivatives. Based on the results it is possible to observe the high nutritional value of the studied oils even after 18?months of conservation.     

Assessment of conventional and microwave heating effects on the variation of the bioactive compounds of Chétoui VOO using HPLC-DAD-ESI-TOF-MS

The goal of this work was to monitor the hydrophilic profile of the virgin olive oil (VOO) of the second main Tunisian variety, “Chétoui”, in order to achieve a better understanding of the behavior of the bioactive phenolic compounds during 0, 2, 5, 10, and 15 min of microwave heating at medium power 800 W, and 0, 2.5, and 5 h of conventional heating at 180 °C. The extent of the oxidative and hydrolytic degradation of the different phenolic subclasses was evaluated using high-performance liquid chromatography (HPLC) coupled to electrospray time-of-flight mass spectrometry (TOF-MS) method. During heating process, the most represented component in Chétoui VOO was found to be isomer 1 and 2 of deacetoxy oleuropein aglycone, and hydroxy decarboxy oleuropein aglycon. These compounds may be considered as direct markers for the degree of transformation of secoiridoids during heating process. Among the studied phenolic compounds, hydroxytyrosol, tyrosol, luteolin and apigenin displayed the highest heating resistance in the whole time range of microwave and conventional applications. However, the main secoiridoids quantified in the fresh VOO, isomer 2 of oleuropein aglycone, and ligstroside aglycone, decreased in concentration with the thermal treatment and this decrease was drastic under conventional heating.     

Volatile compounds in virgin olive oil: occurrence and their relationship with the quality

The stimulation of the human sensory receptors by volatile compounds present in virgin olive oils gives rise to the sensory attributes that describe its delicate and fragrant aroma. The composition of the volatile compounds and their biogenesis is briefly illustrated. Analytical methodologies for evaluating the volatile fraction and the sensory properties of virgin olive oils are elucidated. Compounds responsible for typical flavours are examined and the influence of the main factors on the composition of volatile compounds is discussed. The origin of off-flavours are also described and the consequent changes of volatile composition and of sensory characteristics are analysed. The relationships between volatile compounds and sensory attributes are discussed.     

Evaluation of the oxidative status of virgin olive oils with different phenolic content by direct infusion atmospheric pressure chemical ionization mass spectrometry

Atmospheric pressure chemical ionization mass spectrometry was used to predict the oxidative status of virgin olive oils (VOO) during their storage. VOO samples, with and without phenolic compounds, were stored in the dark at 60 °C up to 7 weeks. The VOO samples were diluted in an alkaline propanol/methanol mixture and directly infused into an ion-trap mass spectrometer. The abundances of the [M−H]⁻ peaks of free fatty acids, oxidized fatty acids, tocopherols and phenolic compounds, jointly with their oxidized forms, were measured and used as predictors. Two linear discriminant analysis (LDA) models were constructed in order to classify samples according to their oxidative levels. The first model was constructed using both VOO samples (with and without phenols), considering as predictors only fatty acids and their oxidized products. The second LDA model was constructed with the VOO sample with phenolic compounds considering as predictors all the peaks measured. In both models, the samples divided in the eight different storage times were correctly classified (100%) by leave-one-out cross-validation with an excellent resolution among all the category pairs (for the first model Wilks’ lambda, λ _w = 0.229 and for the second λ _w = 0.928). This method is a very fast tool for on-line monitoring of VOO oxidation status.     

Effects of conventional heating on the stability of major olive oil phenolic compounds by tandem mass spectrometry and isotope dilution assay

The quality of olive oils is sensorially tested by accurate and well established methods. It enables the classification of the pressed oils into the classes of extra virgin oil, virgin oil and lampant oil. Nonetheless, it would be convenient to have analytical methods for screening oils or supporting sensorial analysis using a reliable independent approach based on exploitation of mass spectrometric methodologies. A number of methods have been proposed to evaluate deficiencies of extra virgin olive oils resulting from inappropriate technological treatments, such as high or low temperature deodoration, and home cooking processes. The quality and nutraceutical value of extra virgin olive oil (EVOO) can be related to the antioxidant property of its phenolic compounds. Olive oil is a source of at least 30 phenolic compounds, such as oleuropein, oleocanthal, hydroxytyrosol, and tyrosol, all acting as strong antioxidants, radical scavengers and NSAI-like drugs. We now report the efficacy of MRM tandem mass spectrometry, assisted by the isotope dilution assay, in the evaluation of the thermal stability of selected active principles of extra virgin olive oil.     

CE- and HPLC-TOF-MS for the characterization of phenolic compounds in olive oil

We present an easy and rapid method for the analysis of phenolic compounds in extra-virgin olive oil by CZE coupled with ESI-TOF-MS. Optimum electrophoretic separation was obtained using a basic carbonate electrolyte. We thus achieved the determination of several important families (phenyl alcohols, phenyl acids, lignans, flavonoids, and secoiridoids) of the polar fraction of the olive oil. Furthermore, other "unknown" compounds were also identified. In addition to the CZE method, HPLC analyses were made, separating compounds belonging to the main families present in this polyphenolic fraction, as well as other new compounds. We compared the results obtained with both techniques and found it was possible to determine more than 45 compounds with both methods. The sensitivity, together with mass accuracy and true isotopic pattern of the TOF-MS, allowed the identification of a broad series of known and so far not described phenolic compounds present in extra-virgin olive oil.     

Maximum-quantum (MaxQ) NMR for the speciation of mixtures of phenolic molecules

The spectroscopic determination of phenolic molecules by means of multiple-quantum (MQ) NMR is demonstrated. Several classes of molecules (simple phenols, flavonols, secoiridoids and lignans) were unambiguously characterized in one pot analysis in an extract of extra virgin olive oil (EVOO).     

A capillary electrophoresis-tandem mass spectrometry methodology for the determination of non-protein amino acids in vegetable oils as novel markers for the detection of adulterations in olive oils

A new analytical methodology based on capillary electrophoresis-mass spectrometry (CE-MS(2)) is presented in this work, enabling the identification and determination of six non-protein amino acids (ornithine, β-alanine, GABA, alloisoleucine, citrulline and pyroglutamic acid) in vegetable oils. This methodology is based on a previous derivatization with butanol and subsequent separation using acidic conditions followed by on-line coupling to an ion trap analyzer for MS(2) detection established through an electrospray-coaxial sheath flow interface. The electrophoretic and interface parameters were optimized obtaining the separation of all compounds in less than 15 min and with resolutions higher than 5. The proposed method was validated by assessing its accuracy, precision (RSD<7% for corrected peak areas), LODs and LOQs (between 0.04-0.19 ng/g and 0.06-0.31 ng/g, respectively) and linearity range (R(2)>0.99), and it was used in order to identify the selected non-protein amino acids in soybean oils, sunflower oils, corn oils and extra virgin olive oils. MS(2) experiments performed the fingerprint fragmentation of these compounds allowing to corroborate ornithine and alloisoleucine in seed oils but not in olive oils. The method was applied to identify and quantify olive oil adulterations with soybean oil detecting in a single run the amino acids in mixtures up to 2% (w/w). The results showed a high potential in using these compounds as novel markers for the detection of adulterations of extra virgin olive oils with seed oils. Thus, the developed method could be considered a simple, rapid and reliable method for the quality evaluation of extra virgin olive oil permitting its authentication.     

Characterization of the Phenolic Fingerprint of Kolovi Extra Virgin Olive Oils from Lesvos with Regard to Altitude and Farming System Analyzed by UHPLC-QTOF-MS

Extra virgin olive oil (EVOO) is recognized for its nutritional virtues and the beneficial health effects deriving from its hydrophilic fraction (phenolic acids, phenolic alcohols, flavonoids, and secoiridoids). The phenolic compounds of EVOOs possess multiple biological properties such as antioxidant, antimicrobial, anticarcinogenic, and anti-inflammatory properties, among others. Considering that EVOOs produced in Greece are recognized as high-quality products due to their rich phenolic content, it is imperative to characterize Greek monovarietal EVOOs and ensure that their uniqueness is closely linked to their botanical and territorial origin. In this work, an ultra-high-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS) analytical method combined with target and suspect screening was used to characterize monovarietal EVOOs of the Kolovi variety from Lesvos, and thereby establish their phenolic fingerprint. Overall, 25 phenols were determined, and the total quantification and semi-quantification results ranged between 251 and 1230 mg/kg, highlighting the high phenolic content of the Kolovi variety from the island of Lesvos in the North Aegean.     

Simple and fast determination of phenolic compounds from different varieties of olive oil by nonaqueous capillary electrophoresis with UV‐visible and fluorescence detection

In this article, we proposed very simple procedures to analyze important phenolic compounds in olive oil samples from different olive varieties. A nonaqueous CE method has been employed. The main phenolic alcohols in virgin olive oil (tyrosol and hydroxytyrosol) and some among the most abundant secoiridoid aglycone derivatives (dialdehydic form of decarboxymethyl elenoic acid linked to hydroxytyrosol, an isomer of oleuropein aglycone and the dialdehydic form of decarboxymethyl elenoic acid linked to tyrosol) were determined by a direct injection into the capillary of the olive oil dissolved in 1‐propanol 1:1 v/v. For the determination of compounds present at lower concentrations, a very simple liquid–liquid extraction method with ethanol has been proposed. The extraction was performed using a relationship 5:1 w/v olive oil/ethanol to achieve the necessary preconcentration of the analytes and the ethanolic extracts were directly injected into the capillary to obtain a very important time reduction. Good recoveries were obtained with both the procedures, using an internal standard. Finally, these procedures were applied to the analysis of these compounds in extra virgin olive oil samples from different varieties of olive.     

Structural characterization of the ligstroside aglycone isoforms in virgin olive oils by liquid chromatography–high‐resolution Fourier‐transform mass spectrometry and H/Dexchange

A systematic structural characterization of the isomeric forms related to ligstroside aglycone (LA), one of the most relevant secoiridoids contained in virgin olive oils, was performed using reverse phase liquid chromatography with electrospray ionization Fourier‐transform single and tandem mass spectrometry, operated in negative ion mode (RPLC‐ESI(?)‐FTMS and FTMS/MS). The high mass resolution and accuracy provided by the adopted orbital trap mass analyzer enabled the recognition of more than 10 different isomeric forms of LA in virgin olive oil extracts. They were related to four different types of molecular structure, two of which including a dihydropyranic ring bearing one or two aldehydic groups, whereas the others corresponded to dialdehydic open‐structure forms, differing just for the position of a C═C bond. The contemporary presence of enolic or dienolic tautomers associated to most of these compounds, stable at room temperature (23°C), was also assessed through RPLC‐ESI‐FTMS analyses operated under H/D exchange conditions, ie, by using D₂O instead of H₂O as co‐solvent of acetonitrile in the RPLC mobile phase. As discussed in the paper, the results obtained for LA indicated a remarkable structural similarity with oleuropein aglycone (OA), the most abundant secoiridoid of olive oil, whose isoforms had been previously characterized using the same analytical approach.     

LC‐DAD‐ESI‐MS/MS–based phenolic profiling and antioxidant activity in Turkish cv. Nizip Yaglik olive oils from different maturity olives

The current study was designed to find out how olive maturity indices (2.5, 3.5, and 4.5) affect the individual phenolic compounds and antioxidant potencies of olive oils produced from cv. Nizip Yaglik olives. Liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry in multiple reaction monitoring mode was utilized for the determination of phenolic composition qualitatively and quantitatively. Findings asserted a quite similar phenolic profile (14 phenols) depending on the various phenolic groups in all oils, while the concentration of total and individual phenolic compounds revealed significant differences between the samples statistically (p < 0.05). Among the individual phenolic classes in all samples, secoiridoids were the most prevailing group and their total content showed a clear significant decline as the olive fruits get ripened. Antioxidant potency values showed a clear diminution attitude during the maturation of the olives. The principal component analysis revealed that oils were discriminated from each other according to phenolic compounds and antioxidant potencies. Moreover, oils obtained from the unripe and medium‐ripe fruits possessed a very good quality marked by their elevated phenolic levels.     

A 2‐D‐HPLC‐CE platform coupled to ESI‐TOF‐MS to characterize the phenolic fraction in olive oil

A 2‐D‐HPLC/CE method was developed to separate and characterize more in depth the phenolic fraction of olive oil samples. The method involves the use of semi‐preparative HPLC (C18 column 250×10 mm, 5 μm) as a first dimension of separation to isolate phenolic fractions from commercial extra‐virgin olive oils and CE coupled to TOF‐MS (CE‐TOF‐MS) as a second dimension, to analyze the composition of the isolated fractions. Using this method, a large number of compounds were tentatively identified, some of them by first time, based on the information concerning high mass accuracy and the isotopic pattern provided by TOF‐MS analyzer together with the chemical knowledge and the behavior of the compounds in HPLC and CE. From these results it can be concluded that 2‐D‐HPLC‐CE‐MS provides enough resolving power to separate hundreds of compounds from highly complex samples, such as olive oil. Furthermore, in this paper, the isolated phenolic fractions have been used for two specific applications: quantification of some components of extra‐virgin olive oil samples in terms of pure fractions, and in vitro studies of its anti‐carcinogenic capacity.     

Colloidal structures in natural oils

Natural oils are in general mixtures of triglycerides of various fatty acids containing also small quantities of a multitude of other molecules, the minor components. Their composition depends on the type of their sources, either vegetable or animal, and on agronomic and climatic conditions. Extraction process highly influences the final composition of the natural oils. The bulk triglyceride phase presents some structural order due to the internal polarity of these molecules. The presence of the minor components, which are either amphiphilic or even hydrophilic together with some remaining moisture, induces colloidal association within the lipophilic triglyceride phase. These local organizates host proteins and antioxidants that preserve their activity influencing the quality and stability of the edible oils. Of specific interest is the case of the veiled virgin olive oil, the untreated juice of the olives. Veiled virgin olive oil is unique among vegetable oils that is consumed without any refining process and consequently possesses colloidal structure. It is remarkable that this unfiltered oil presents higher resistance to oxidation than the filtered ones, indicating the significance of internal structure to the final quality of the product.