Instrumental and multivariate statistical analyses for the characterisation of the geographical origin of Apulian virgin olive oils

In this paper, virgin olive oils (VOOs) coming from three different geographic origins of Apulia, were analysed for free acidity, peroxide value, spectrophotometric indexes, chlorophyll content, sterol, fatty acid, and triacylglycerol compositions. In order to predict the geographical origin of VOOs, different multivariate approaches were applied. By performing principal component analysis (PCA) a modest natural grouping of the VOOs was observed on the basis of their origin, and consequently three supervised techniques, i.e., general discriminant analysis (GDA), partial least squares-discriminant analysis (PLS-DA) and soft independent modelling of class analogy (SIMCA) were used and the results were compared. In particular, the best prediction ability was produced by applying GDA (average prediction ability of 82.5%), even if interesting results were obtained also by applying the other two classification techniques, i.e., 77.2% and 75.5% for PLS-DA and SIMCA, respectively.     

Geographical classification of Greek virgin olive oil by non-parametric multivariate evaluation of fatty acid composition

Fatty acid compositional data for Greek virgin olive oils from 24 years of harvest and various regions and cultivars were evaluated using chemometric methods. Non-parametric discriminant analysis after proper transformation of the data seems to be a suitable approach to characterise the oils according to the geographical origin and may produce a scientific basis for the assignment of an ‘appellation d'origine’ trade mark.     

Multivariate analysis of NMR fingerprint of the unsaponifiable fraction of virgin olive oils for authentication purposes

A new approach to the geographical characterisation of virgin olive oils (VOOs) based on the ¹H NMR fingerprint of the unsaponifiable matter is presented. The ¹H NMR spectra of the unsaponifiable fraction of virgin olive oils from Spain, Italy, Greece, Tunisia, Turkey, and Syria were analysed by several pattern recognition techniques (LDA, PLS-DA, SIMCA, and CART). PLS-DA (PLS-1 approach) obtained the best classification results for all classes. Moreover, ¹H NMR spectra of the bulk oil, and its corresponding unsaponifiable fraction, as well as the subfractions of the unsaponifiable fraction (alcohol, sterol, hydrocarbon, and tocopherol fractions) were studied in the search for the markers that multivariate techniques revealed to be related to the geographical origin of olive oils. Additionally, a preliminary study regarding ¹H NMR data of the bulk oil and the corresponding unsaponifiable fraction of VOOs suggested that these spectral data contained complementary information for the geographical characterisation of VOOs.     

Study of oils from Calabrian olive cultivars by chemometric methods

A study of characterisation of a typical Italian food is performed by chemometrics. The olive oils from some cultivars of Calabria have been characterised according to their origin and olive genotype using the chemical information mainly provided by 14 chemical parameters of virgin olive oils. In particular, the models of three cultivars of Calabria (Carolea, Cassanese, Dolce di Rossano) were deeply studied. The microclimate of this region has a lower influence on fatty acid composition than the genotype. Using simple and relatively inexpensive analytical parameters as fatty acids and chemometric techniques it has been possible to characterise and classify the olive oils (60–94% prediction rate).     

Stability of virgin olive oil and behaviour of its natural antioxidants under medium temperature accelerated storage conditions

The aim of the present research is to advance the study of virgin olive oil stability, in particular the behaviour of its natural antioxidants during long term storage. The changes undergone by tocopherols and complex and simple phenolic compounds and their antioxidant activity were studied under medium temperature (50 °C) accelerated oxidation conditions over a storage period of 8 months. Different oxygen availability (open and closed bottles, OB and CB) and four different monovarietal Tunisian virgin olive oils (VOO; Chemlali, Chétoui, El Hor and Oueslati) which vary in their fatty acid profile and content of natural antioxidants were employed. Oueslati VOO showed the lowest initial oxidation rate (4.6 PV/week) and as a consequence the highest induction period (IP; 16 days) as compared to the other VOOs studied (initial rate from 7.7 to 12.1 PV/week and IP from 9 to 12 days for El Hor and Chemlali, respectively, in both cases). In all cases the complete depletion of α-tocopherol corresponded with the observed induction period. Since the oxidation rate in the varieties studied is not sufficiently different to explain the great differences in the initial degradation rate observed in the phenolic compounds (mainly between Chétoui VOO, 163 μmol/kg/week, as compared to the other VOO oils, e.g. 6.1 and 2.5 μmol/kg/week, respectively, in Chemlali and El Hor VOOs), a concentration dependent degradation rate of complex phenols is suggested.     

High performance size-exclusion chromatography analysis of polar compounds applied to refined, mild deodorized, extra virgin olive oils and their blends: An approach to their differentiation

An investigation was carried out to evaluate the use of High Performance Size-Exclusion Chromatography (HPSEC) of polar compounds of refined, mild deodorized, extra virgin olive oils as well as of their blends, in attempting to reveal significant differences in the amounts of the substance classes constituting polar compounds among these oils. Two sets of blends were prepared by mixing an extra virgin olive oil with both refined and mild deodorized olive oils in increasing amounts. The obtained data highlighted that the triacylglycerol oligopolymers were absent or present in traces in the extra virgin olive oil, while their mean amount was equal to 0.04 g/100 g and 0.72 g/100 g in mild deodorized and refined olive oils, respectively. Oxidized triacylglycerols and diacylglycerols were more abundant in mild deodorized oil and refined oil than in extra virgin olive oil. The Factorial Discriminant Analysis of the data showed that the HPSEC analysis could reveal the presence of refined/mild deodorized oils in extra virgin olive oils. In particular, the classification functions obtained allowed designation of mixtures containing at least 30 g/100 g of mild deodorized oil and all those containing refined olive oil as deodorized oil, therefore as oils subjected to at least a mild refining treatment.     

Implementation of physicochemical and sensory analysis in conjunction with multivariate analysis towards assessing olive oil authentication/adulteration

The authenticity of products labeled as olive oils, and in particular as virgin olive oils, stands for a very important issue both in terms of its health and commercial aspects. In view of the continuously increasing interest in virgin olive oil therapeutic properties, the traditional methods of characterization and physical and sensory analysis were further enriched with more advanced and sophisticated methods such as HPLC-MS, HPLC-GC/C/IRMS, RPLC-GC, DEPT, and CSIA among others. The results of both traditional and "novel" methods were treated both by means of classical multivariate analysis (cluster, principal component, correspondence, canonical, and discriminant) and artificial intelligence methods showing that nowadays the adulteration of virgin olive oil with seed oil is detectable at very low percentages, sometimes even at less than 1%. Furthermore, the detection of geographical origin of olive oil is equally feasible and much more accurate in countries like Italy and Spain where databases of physical/chemical properties exist. However, this geographical origin classification can also be accomplished in the absence of such databases provided that an adequate number of oil samples are used and the parameters studied have "discriminating power."     

Microwave heating effects on the chemical composition and the antioxidant capacity of tataouine virgin olive oils from Tunisia

Four Tunisian virgin olive oils (VOOs), derived from varieties (Chemlali Tataouine, Zarrazi Douirat, Fakhari Douirat, and Dhokar Douirat) grown in the harsh pedoclimatic conditions of the region of Tataouine, were evaluated for their responses to microwave heating. Aside from fatty acid composition, all other evaluated parameters were affected by microwave heating, and their variations depend on the genetic factor. Chemlali Tataouine VOO exhibited the slowest biophenol degradation rate and the least diminution in oxidative stability and consequently, its total fraction and both lipidic and methanolic fractions remained unchanged with an exceptional antioxidant potential. In the remaining studied VOOs, the biophenol contents, the oxidative stability, and the antioxidant potential underwent gradual decreases; nevertheless, their levels at the longer treatment time are close to some fresh VOOs. These results should be taken into consideration when Tataouine VOOs are recommended for microwave heating.     

Evaluation of the quality of olive oil using fatty acid profiles by direct infusion electrospray ionization mass spectrometry

Electrospray ionization mass spectrometry is used to predict the olive oil quality according to European Union marketing standards. Samples were 1:50 diluted in an alkaline 85:15 (v/v) propanol/methanol mixture and directly infused into the electrospray ionization source of an ion trap mass spectrometer. The establishment of ratios of the peak abundances of the free fatty acids followed by linear discriminant analysis was employed to predict the olive oil quality grade. In addition, using multiple linear regression and partial least-squares regression, the percentages of extra virgin and virgin olive oils in binary mixtures were predicted with 5–11% average prediction errors.     

Analytical evaluation of two monovarietal virgin olive oils cultivated in the south of Tunisia: Jemri‐Bouchouka and Chemlali‐Tataouin cultivars

BACKGROUND: The characterisation of virgin olive oils from two Tunisian cultivars, growing in the Tataouin zone, namely Jemri‐Bouchouka, a rare olive cultivar, and Chemlali‐Tataouin, was carried out. Several analytical parameters were evaluated; these include quality index, fatty acids, phenolic, chlorophyll, carotenoid, squalene, α‐tocopherol compositions and oxidative stability. RESULTS: Jemri‐Bouchouka olive oil had the highest value of oleic acid (74.50%) while Chemlali‐Tataouin was characterised by a high percentage of palmitic acid (14.75%), which makes this oil freeze at a low temperature. On the other hand, Jemri‐Bouchouka oil was characterised by a low phenolic and α‐tocopherol content (267.72 mg GAE kg^?1 and 278.34 mg kg^?1, respectively). Ten phenolic compounds were identified. The main phenols found in the two olive oils were oleuropein aglycon and pinoresinol. All phenolic compounds showed significant correlations with oxidative stability. CONCLUSION: The analytical parameters of virgin olive oil that were determined in this study were greatly influenced by cultivar. © 2012 Society of Chemical Industry     

FTIR Spectroscopy and PLS-DA Classification and Prediction of Four Commercial Grade Virgin Olive Oils from Morocco

Fourier transform infrared (FTIR) coupled to chemometrics was shown to be a useful method to classify and predict the quality of four commercial grade virgin olive oils (VOO). FTIR and physicochemical data were collected using a set of 70 samples representing extra virgin (EV), virgin (V), ordinary virgin (OV), and lampante (L) commercial grade olive oils collected in Beni Mellal region (central Morocco). Two partial least squares discriminant analysis (PLS-DA) models using physicochemical data and FTIR data were established and compared. The PLS-DA model using only physicochemical data was not accurate enough to distinguish satisfactorily among OV, V, and EV olive oil grades. On the contrary, the PLS-DA model on FTIR data was better in the calibration, able to describe 98 % of the spectral information and predicting 93 % of the VOO grades. In the external validation, this PLS-DA model accurately classified VOO commercial grades with prediction accuracy of 100 %. The proposed procedure is fast, nondestructive, simple, and easy to operate, and it is recommended for the quick monitoring of olive oil’s quality.     

MALDI-TOF mass spectrometry detection of extra-virgin olive oil adulteration with hazelnut oil by analysis of phospholipids using an ionic liquid as matrix and extraction solvent

The adulteration of extra virgin olive oil (EVOO) with hazelnut oil (HO) is frequent and constitutes a serious concern both for oil suppliers and consumers. The high degree of similarity between the two oils as regards triacylglycerol, total sterol and fatty acid profile, complicates the detection of low percentages of HO in EVOO. However, phospholipids (PLs) are usually present in seed oils at a concentration range of 10–20 g/kg, while the amounts of PLs in VOOs are 300–400 times lower. Thus, in this work a sample pretreatment procedure focused towards the selective PLs extraction was developed; the Bligh–Dyer extraction procedure was modified introducing the ionic liquid resulting from the combination of TBA (tributylamine) and CHCA (α-Cyano-4-hydroxycinnamic acid) as extraction solvent. The selective extraction and enrichment of phospholipids from EVOO and HO samples was then achieved. The relevant extracts were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) using the same ionic liquid TBA-CHCA as MALDI matrix, that was found to be very suitable for PLs analysis. In fact, a remarkable increase of the phospholipids signals, with a simultaneous decrease of those relevant to triacylglycerols and diacylglycerols, was observed in the relevant mass spectra. The applicability of the whole method to the individuation of the presence of HO in EVOO was demonstrated by the analysis of EVOO samples progressively adulterated with variable quantities of HO, that was still detectable at a 1% contamination level.     

Pigments profile in monovarietal virgin olive oils from various Italian olive varieties

This paper presents the investigation of the chlorophyll and carotenoid pigments composition in monovarietal virgin olive oils produced from the five main olive varieties (Minuta, Ottobratica, Calabrese, Ogliarola, Baddarica) cultivated in Sicily (southern Italy), from four main olive varieties (DolceAgogia, Moraiolo, Leccino, Frantoio) cultivated in Umbria (central Italy), and from three main olive varieties (Leccino, Oliva Nera di Collecorto, Noccioluta) cultivated in Molise (central Italy). Reversed-phase high performance liquid chromatography using a C-30 column with photodiode array detection was used for pigments analyses. In all, 19 compounds were identified and quantified in 60 olive oils samples. The qualitative pigments pattern was similar among the varieties investigated, whereas quantitative differences were found among the different cultivars; among the varieties investigated in this work, the oils from Umbria showed the highest pigment content (34.19 ppm in average), followed by the oils from Molise (18.61 ppm in average) and the oils from Sicily which showed the lowest pigment contents (13.38 ppm in average). In general, pheophytin a was the major component (range 0.49–19.42 ppm), followed by β-carotene (range 1.27–9.30 ppm) and lutein (range 0.44–5.12 ppm). Those differences may be due to genetic factors and/or geographical differences. Moreover, auroxanthin was detected for the first time in olive oils and was detected only in olive oils from Umbria and Molise regions. The ratio between the two isochromic pigment fractions, namely the ratio between the chlorophyll and carotenoid fractions showed an average value close to unity. The lutein/β-carotene ratio was less than one in the majority of the cases. These parameters, along with other analytical parameters, could be used as indicators of typicality in olive oils. The presence of a specific pigment profile in olive oils could infact be used to guarantee the genuineness of the product, since the quality control of food requires a precise knowledge of the pigments composition of the original products.     

Evaluating the potential of phenolic profiles as discriminant features among extra virgin olive oils from Moroccan controlled designations of origin

Herewith, the potential of an approach based on the combination of the determination of phenolic compounds and the use of chemometric treatments has been evaluated to establish, for the first time, promising models to authenticate the provenance of Moroccan monovarietal olive oils produced under different geographical origin indication systems. To achieve this purpose, 136 commercial extra virgin olive oil samples from three diverse production areas (Meknès territory; the Protected Geographical Indication Ouazzane; and the Protected Designation of Origin Tyout-Chiadma) were collected over two consecutive crop seasons (2012/2013 and 2013/2014). Their phenolic fraction composition was investigated by using high performance liquid chromatography coupled to mass spectrometry (HPLC-ESI-IT MS). The results showed that geographical provenance and harvest season had a marked influence on the content of identified phenolic compounds. Principal components analysis (PCA) and linear discriminant analysis (LDA) were applied to test the potential of the determined compounds as geographical discriminant features, achieving a noticeable discrimination among the three evaluated regions. The contribution of each analyte to the statistic model has been evaluated in depth.     

Sterolic composition of Chétoui virgin olive oil: Influence of geographical origin

The sterol profile of Tunisian virgin olive oils produced from Chétoui cultivar, the second main variety cultivated in the north of the country, grown under different environmental conditions, was established by gas chromatography using a flame ionisation detector. More than ten compounds were identified and characterised. As expected for virgin olive oil, the main sterols found in all Chétoui olive oils were β-sitosterol, Δ5-avenasterol, campesterol and stigmasterol. Cholesterol, 24-methylenecholesterol, clerosterol, campestanol, sitostanol, Δ7-stigmastenol, Δ5,24-stigmastadienol, and Δ7-avenasterol were also found in all samples, but in lower amounts. Most of these compounds are significantly affected by the geographical origin. The majority of the Chétoui virgin olive oils analysed respected EC Regulation No. 2568, and in all cases total sterol amounts were higher than the minimum limit set by legislation, ranging from 1017 to 1522 mg/kg.     

Pigments composition in monovarietal virgin olive oils from various sicilian olive varieties

This paper presents the first investigation of the chlorophyll and carotenoid pigments composition in sicilian monovarietal virgin olive oils from the three (Cerasuola, Nocellara, Biancolilla) main olive varieties cultivated in Sicily (Italy). In all, 19 compounds were identified and quantified in 24 olive oil samples. The application of reversed-phase liquid chromatography with photodiode array detection using a C-30 column in the simultaneous qualitative–quantitative analysis of virgin olive oils pigments, has been shown. The qualitative pigment pattern was similar among the varieties investigated, whereas quantitative differences were found among the different cultivars, which can all be considered as having an high pigment content. Pheophytin a, was the major component (19.36–25.04 ppm), followed by β-carotene (8.06–16.27 ppm). Pheophytin a′ (2.92–4.17 ppm), lutein (2.28–4.49 ppm) and neoxanthin (1.54–2.11 ppm) were also well represented. The presence of carotenoid esters was also detected. The neoxanthin and β-carotene contents were higher compared to reports present in the literature for other olive oil varieties. This may be due to genetic factors and/or geographical differences. The ratio between the two isochromic pigment fractions, namely the chlorophyll and the carotenoid fractions, was around one in all varieties, showing that they were in balance. The lutein/β-carotene ratio was less than one in all cases. These parameters, along with other analytical parameters, could be used as indicators of typicality in olive oils. The presence of a specific pigment profile in olive oils could in fact be used to guarantee the genuineness of the product, since the quality control of food requires a precise knowledge of the pigments composition of the original products.     

A SNP-based PCR–RFLP capillary electrophoresis analysis for the identification of the varietal origin of olive oils

Authenticity and traceability of high quality monovarietal extra virgin olive oils is a major concern for markets and consumers. Although analytical chemistry techniques are widely used to satisfy these needs recently developed DNA-based methods can serve as complementary approaches. A SNP database comprising 10 Greek olive varieties was constructed and five SNPs, residing in restriction sites, were selected for the development of a PCR–RFLP capillary electrophoresis method to discriminate these varieties using leaf DNA as template. An identification key was constructed indicating that five SNPs were adequate to discriminate nine out of the 10 varieties. As a proof of principle the assay was applied on DNA extracted from five of their corresponding monovarietal olive oils. Three SNPs were able to identify the varietal origin of these olive oils confirming the validity of this approach.     

Classification of different quality virgin olive oils by metal-oxide sensors

The European Union classifies virgin olive oils into three categories, extra virgin, virgin and lampante; lampante being the only oil that cannot be consumed before undergoing a refining process. A mathematical model based on two metal-oxide sensors has been designed and checked in order to detect on-line lampante oils inside the production systems. The model was developed using 114 samples and was successfully tested with an external test set of 55 samples taken from different single varietal olive oils and geographical origins. The model was able to detect 100% of non-lampante virgin olive oils and 89.5% of lampante virgin olive oils.