Temperature Dependence of Oxidation Kinetics of Extra Virgin Olive Oil (EVOO) and Shelf-Life Prediction.

Producers have to guarantee the extra virgin olive oil (EVOO) quality characteristics reported in the Regulation (CEE) 2568/91 throughout the product shelf-life (SL). Unfortunately, due to the development of oxidative reactions, some quality indices change during storage leading to a progressive deterioration of EVOO quality. To avoid the risk of product downgrading in the virgin oil category, the development of effective shelf-life prediction models is extremely important for the olive oil industry. In this research, the accelerated shelf-life testing (ASLT) protocol was applied to evaluate the temperature dependence of selected oxidation indexes as well as to develop a shelf-life predictive model. The evolution of conventional (peroxide value, K232, K270, polyphenols, tocopherols and hexanal) and unconventional parameters (conjugated trienes and pyropheophytin a) was monitored in bottled EVOO stored in the dark at increasing temperature (25, 40, 50 and 60 °C). Accordingly, for well-packed products with reduced oxygen in headspace, the best shelf-life index allowing the ability to predict EVOO SL turned out to be K270. In addition, pyropheophytin a (%) has been shown to be more sensitive to temperature changes than the secondary oxidation indices, thus suggesting its use as a freshness indicator for storage temperatures higher than 25 °C.

Direct determination of 3-chloropropanol esters in edible vegetable oils using high resolution mass spectrometry (HRMS-Orbitrap).

A series of refined edible oils derived from mixed seeds, peanuts, corn, sunflower and palm obtained from the local supermarket were analyzed for their content of 3-MCPD esters. A direct analytical method for the determination of 3-monochloropropane-1,2-diol esters (3-MCPD esters) was applied to investigate the major MCPD esters found in common edible oils; in particular seven types of monoesters and eleven types of diesters were detected. The limits of detection (LODs) for monoesters and diesters of 3-MCPD were in the range of 0.079-12.678 µg kg-1 and 0.033-18.610 µg kg-1 in edible oils, and the ranges of limits of quantitation (LOQs) were 0.979-38.035 µg kg-1 and 0.100-55 µg kg-1, respectively. The recoveries of 3-MCPD esters from oil samples were in the range of 80-100%, with RSD ranging between 1.9 and 11.8%. The concentration levels of total 3-MCPD diesters in vegetable oil samples were in the range from 0.106 up to 3.444 µg g-1 whereas total monoesters ranged from 0.005 up to 1.606 µg g-1.

Characterisation of minor components in vegetable oil by comprehensive gas chromatography with dual detection.

The profile of minor compounds, such as alcohols, sterols, free and alkyl fatty acids, waxes, etc., was investigated in different vegetable oils by a comprehensive gas chromatographic system, coupled with a simultaneous dual detection (flame ionisation detector and mass spectrometer) for quantitative and qualitative purposes. Such a system generated a unique two-dimensional chromatogram to be used as a chemical fingerprint. Multi-level information, due not only to a more "comprehensive" preparation technique, but also thanks to the exploitation of a more powerful and sensitive analytical determination allowed the extrapolation of diagnostic information from the minor components profile of different vegetable oils, along with their characteristic profile. Furthermore, an admixture of an extra virgin olive oil with a low amount of sunflower and palm oils was evaluated, attesting to the powerful diagnostic information provided by the proposed approach.

Rapid direct analysis to discriminate geographic origin of extra virgin olive oils by flash gas chromatography electronic nose and chemometrics.

At present, the geographical origin of extra virgin olive oils can be ensured by documented traceability, although chemical analysis may add information that is useful for possible confirmation. This preliminary study investigated the effectiveness of flash gas chromatography electronic nose and multivariate data analysis to perform rapid screening of commercial extra virgin olive oils characterized by a different geographical origin declared in the label. A comparison with solid phase micro extraction coupled to gas chromatography mass spectrometry was also performed. The new method is suitable to verify the geographic origin of extra virgin olive oils based on principal components analysis and discriminant analysis applied to the volatile profile of the headspace as a fingerprint. The selected variables were suitable in discriminating between "100% Italian" and "non-100% Italian" oils. Partial least squares discriminant analysis also allowed prediction of the degree of membership of unknown samples to the classes examined.

Reliability of the ΔECN42 limit and global method for extra virgin olive oil purity assessment using different analytical approaches.

Two data elaboration approaches for evaluating olive oils authenticity were compared: (I) determination of the difference between the theoretical and actual amounts of triacylglycerols with partition number 42 (ΔECN42 ⩽ |0.2|); and (II) the global method, which considers also partition numbers 44 and 46 (returning a "correct"/"not correct" result). Analysis of 31 genuine extra virgin olive oil samples was performed using different analytical methods, namely liquid chromatography (LC) coupled with a refractive index detector (RID) and LC coupled with a mass spectrometry (MS), and the results compared. Several false positives were highlighted using the ΔECN42 limit with both instrumental approaches. The global method algorithm returned "correct" results for all the samples analysed (except two that gave no results) with LC-MS; on the other hand, 10 false positives were obtained elaborating data deriving from NARP-LC-RID analysis.

Fingerprinting of vegetable oil minor components by multidimensional comprehensive gas chromatography with dual detection.

The potentiality of a multidimensional comprehensive gas chromatographic (GC × GC) method, employing a simultaneous dual detection (FID and mass spectrometer), to generate peculiar two-dimensional chromatograms to be used as a chemical fingerprint, was investigated to characterize minor compounds in edible oil, particularly olive oil. The best column combination for this application was investigated comparing two column sets (orthogonal or reverse-type), equivalent in terms of theoretical plate number, but differing in stationary phase combination. The apolar × mid-polar set gave a superior separation power, thus was used for further characterization. Different levels of information were extrapolated from the two-dimensional chromatogram. Using the FID, reliable quantification of the alkyl esters fatty acids and waxes was obtained, comparable to the results obtained using the official method, as required by the European legislation. However, thanks to a slight modification of the sample preparation method, the increased separation power obtained using the GC × GC method, and the support of the mass spectrometer detector, further diagnostic information was extrapolated considering the free sterol and tocopherol fractions. In particular, the profiles of extra virgin olive oil samples were compared with a hazelnut oil sample, highlighting that the latter was characterized by a larger number of compounds, completely absent in the extra virgin olive oil samples, which can be used to detect illegal admixtures.

Toward a definition of blueprint of virgin olive oil by comprehensive two-dimensional gas chromatography.

This study investigates the applicability of an iterative approach aimed at defining a chemical blueprint of virgin olive oil volatiles to be correlated to the product sensory quality. The investigation strategy proposed allows to fully exploit the informative content of a comprehensive multidimensional gas chromatography (GC×GC) coupled to a mass spectrometry (MS) data set. Olive oil samples (19), including 5 reference standards, obtained from the International Olive Oil Council, and commercial samples, were submitted to a sensory evaluation by a Panel test, before being analyzed in two laboratories using different instrumentation, column set, and software elaboration packages in view of a cross-validation of the entire methodology. A first classification of samples based on untargeted peak features information, was obtained on raw data from two different column combinations (apolar×polar and polar×apolar) by applying unsupervised multivariate analysis (i.e., principal component analysis-PCA). However, to improve effectiveness and specificity of this classification, peak features were reliably identified (261 compounds), on the basis of the MS spectrum and linear retention index matching, and subjected to successive pair-wise comparisons based on 2D patterns, which revealed peculiar distribution of chemicals correlated with samples sensory classification. The most informative compounds were thus identified and collected in a "blueprint" of specific defects (or combination of defects) successively adopted to discriminate Extra Virgin from defected oils (i.e., lampante oil) with the aid of a supervised approach, i.e., partial least squares-discriminant analysis (PLS-DA). In this last step, the principles of sensomics, which assigns higher information potential to analytes with lower odor threshold proved to be successful, and a much more powerful discrimination of samples was obtained in view of a sensory quality assessment.

Rapid screening of fatty acid alkyl esters in olive oils by time domain reflectometry.

The main aim of the present research is to assess the possibility of quickly screening fatty acid alkyl esters (FAAE) in olive oils using time domain reflectometry (TDR) and partial least-squares (PLS) multivariate statistical analysis. Eighteen virgin olive oil samples with fatty acid alkyl ester contents and fatty acid ethyl ester/methyl ester ratios (FAEE/FAME) ranging from 3 to 100 mg kg(-1) and from 0.3 to 2.6, respectively, were submitted to tests with time domain resolution of 1 ps. The results obtained in test set validation demonstrated that this new and fast analytical approach is able to predict FAME, FAEE, and FAME + FAEE contents with R(2) values of 0.905, 0.923, and 0.927, respectively. Further measurements on mixtures between olive oil and FAAE standards confirmed that the prediction is based on a direct influence of fatty acid alkyl esters on the TDR signal. The suggested technique appeared potentially suitable for monitoring one of the most important quality attribute of the olive oil in the extraction process.

Direct-immersion solid-phase microextraction coupled to fast gas chromatography mass spectrometry as a purification step for polycyclic aromatic hydrocarbons determination in olive oil.

The aim of the present work was to optimize a preparation step for polycyclic aromatic hydrocarbons in a fatty extract. Solid-phase microextraction is an easy preparation technique, which allows to minimize solvent consumption and reduce sample manipulation. A Carbopack Z/polydimethylsiloxane fiber, particularly suitable for extraction of planar compounds, was employed to extract polycyclic aromatic hydrocarbons from a hexane solution obtained after a previous extraction with acetonitrile from oil, followed by a liquid-liquid partition between acetonitrile and hexane. The proposed method was a rapid and sensitive solution to reduce the interference of triglycerides saving the column life and avoiding frequent cleaning of the mass spectrometer ion source. Despite the non-quantitative extraction of polycyclic aromatic hydrocarbons from oil using acetonitrile, the signal-to-noise ratio was significantly improved obtaining a limit of detection largely below the performance criteria required by the European Union legislation.

Influence of phenols mass fraction in olive (Olea europaea L.) paste on volatile compounds in Buza cultivar virgin olive oil.

The influence of the phenolic content in olive paste of cv. Buza increased by the addition of an aqueous solution of phenolic extract of freeze-dried olive pulp (cv. Istarska bjelica) on the final products of the lipoxygenase pathway in oil was studied. Increases by 12, 38, and 56% for ripe fruits (maturity index = 4.0) and by 38% for unripe fruits (maturity index = 1.2) were examined. Phenols in the olive paste were determined according to the HPLC method, whereas volatiles in oil were determined according to SPME-GC-MS. A significant negative effect on Z-3-hexenal and E-2-hexen-1-ol (Tukey's test, p < 0.05) was found for ripe fruits (average decreases of 55 and 60%, respectively), but not for the unripe sample. Positive effects in both ripening levels were found for Z-3-hexenyl acetate (average increase of 68% for ripe and a double increase for unripe fruits) and total C5 compounds (average increase of 32% for ripe and an increase of 30% for unripe fruits).

Detailed elucidation of hydrocarbon contamination in food products by using solid-phase extraction and comprehensive gas chromatography with dual detection.

The present work is focused on the development/optimization of a comprehensive two-dimensional gas chromatography method, with dual detection [flame ionization (FID) and mass spectrometric], for the simultaneous identification and quantification of mineral-oil contaminants in a variety of food products. The two main classes of contaminants, namely saturated and aromatic hydrocarbons, were previously fractionated on a manually-packed silver silica solid-phase extraction (SPE) cartridge. The quantitative results were compared with those obtained by performing a large volume injection, in a GC-FID system, after the same SPE process and by an on-line liquid-gas chromatography method, with very similar results observed. The presence of a series of unknown compounds, that appeared when using the off-line methods, was investigated using the mass spectrometric data, and were tentatively-identified as esterified fatty acids, most probably derived from vegetable oil based ink.

Microwave-assisted extraction of edible Cicerbita alpina shoots and its LC-MS phenolic profile.

BACKGROUND: Crude extracts obtained from the edible shoots of Cicerbita alpina using microwave-assisted extraction have been qualitatively profiled by liquid chromatography coupled with an ion trap mass spectrometry detector and an electrospray ionization interface (LC/ESI-MS(3) ) for their phenolic content. The main challenge of the present investigation was to create a working strategy designed to obtain a rich phenolic profile despite the limited amount of starting plant material and phytochemical data available. RESULTS: The best extraction conditions (temperature 90 °C; time 5 min; solvent methanol:water 50:50; sample weight 3 g) were achieved using a full factorial 2(4) experimental design. Fifteen compounds, including flavonoid conjugates and phenolic acid derivatives, were detected and tentatively identified. The total phenolic content varied from 93.58 mg g(-1) gallic acid equivalents (GAE), for the cultivated plant to 10.54 mg g(-1) GAE for the wild one, whereas the total flavonoid content varied from 145.00 mg g(-1) rutin for the cultivated plant to 25.22 mg g(-1) rutin for the wild one. CONCLUSION: A total of 11 compounds are herein reported, for the first time, as coming from this plant source.

A rapid multidimensional liquid-gas chromatography method for the analysis of mineral oil saturated hydrocarbons in vegetable oils.

The present paper describes an investigation directed toward the development of a rapid heart-cutting LC-GC method for the analysis of mineral oil saturated hydrocarbons contained in vegetable oils. The automated LC-GC experiments were carried out by using a system equipped with a syringe-type interface, capable of both heart-cutting and comprehensive (LC × GC) two-dimensional analysis. The first dimension separation was achieved on a 100 mm × 3 mm ID × 5 µm d(p) silica column, operated under isocratic conditions (hexane). A single 30-s cut, corresponding to a 175 µL volume, was transferred to a programmed temperature vaporizer. After the large volume injection, the target analytes were separated in a rapid manner (~9 min) using a 15 m × 0.1mm ID × 0.1 µm micro-bore GC capillary. The overall LC-GC run time enables the analysis of ca. 4 samples/hour. Quantification was performed by using external calibration, in the 1-200 mg/kg range. The method was validated in terms of linearity, precision, limits of detection and quantification, and accuracy. A series of commercial samples were subjected to analysis. Various degrees of contamination were found in all samples, in the 7.6-180.6 mg/kg range.

Visible and near-infrared absorption spectroscopy by an integrating sphere and optical fibers for quantifying and discriminating the adulteration of extra virgin olive oil from Tuscany.

Because of its high price, extra virgin olive oil is frequently targeted for adulteration with lower quality oils. This paper presents an innovative optical technique capable of quantifying and discriminating the adulteration of extra virgin olive oil caused by lower-grade olive oils. An original set-up for diffuse-light absorption spectroscopy in the wide 400-1,700 nm spectral range was experimented. It made use of an integrating sphere containing the oil sample and of optical fibers for illumination and detection; it provided intrinsically scattering-free absorption spectroscopy measurements. This set-up was used to collect spectroscopic fingerprints of authentic extra virgin olive oils from the Italian Tuscany region, adulterated by different concentrations of olive-pomace oil, refined olive oil, deodorized olive oil, and refined olive-pomace oil. Then, a straightforward multivariate processing of spectroscopic data based on principal component analysis and linear discriminant analysis was applied which was successfully capable of predicting the fraction of adulterant in the mixture, and of discriminating its type. The results achieved by means of optical spectroscopy were compared with the analysis of fatty acids, which was carried out by standard gas chromatography.

Analysis of fresh and aged tea tree essential oils by using GCxGC-qMS.

The present research is focused on the qualitative elucidation of the chemical profile of fresh tea tree essential oil and an oxidized, aged (circa 1984) counterpart by using the most powerful analytical tool available today for volatile analysis, namely comprehensive two-dimensional gas chromatography (GCxGC) in combination with a mass spectrometer (MS). The rapid-scanning quadrupole (q) MS system employed generated a sufficient number of spectra/s (20/s) for the reliable identification of the high-speed GCxGC peaks. The total ion current GCxGC-qMS chromatogram of the fresh product was characterized by the presence of approximately 130 unknowns. Among these, 61 peaks were assigned with spectral similarities > or = 90%, while 28 components presented MS library matches in the 80-89% range. With regards to the oxidized essential oil, about 180 volatiles were visible on the 2D plane with 63 of these characterized by library "hits" > or = 90% and 45 presenting similarities within the 80-89% range. The use of the enhanced-resolution 3D methodology enabled the full separation of the samples analyzed and, hence, a clear distinction between the essential oils.

Characterization of the yerba mate (Ilex paraguariensis) volatile fraction using solid-phase microextraction-comprehensive 2-D GC-MS.

The present research is focused on the use of a solid-phase microextraction-comprehensive 2-D GC methodology, in the analysis of the volatile fraction of yerba mate. Yerba mate is used for the generation of a tea-like beverage, widely consumed in South America. A rapid-scanning quadrupole mass spectrometer (qMS), employed as a detection system and operated at a 25 Hz scanning frequency, supplied high-quality mass spectra. The effectiveness of the 3-D comprehensive 2-D GC-qMS experiment was compared to that of GC-qMS analysis on the same sample. Peak identification, in both applications, was achieved through MS library matching, with the support of linear retention index data. Apart from a great increase in the number of analytes separated (approx. by a factor of 5) and identified (approx. by a factor of 3.5), the comprehensive 2-D GC-qMS approach enabled the determination of a high number of hazardous contaminants (aliphatic hydrocarbons, polycyclic aromatic hydrocarbons, and plasticizers), barely visible in the GC-qMS analysis.

Enhanced resolution comprehensive two-dimensional gas chromatography applied to the analysis of roasted coffee volatiles.

The present research is based on the full exploitation of the separation power of a 0.05 mm internal diameter (ID) capillary, as a comprehensive two-dimensional (2D) GC (GC x GC) secondary column, with the objective of attaining very high-resolution second dimension separations. The aim was achieved by using a split-flow system developed in previous research [P.Q. Tranchida, A. Casilli, P. Dugo, G. Dugo, L. Mondello, Anal. Chem. 79 (2007) 2266], and a dual-oven GC x GC instrument. The column combination employed consisted of a polar 30 m x 0.25 mm ID column connected, by means of a T union, to a detector-linked high-resolution 1.1 m x 0.05 mm ID apolar analytical column and to a 0.33 m x 0.05 mm ID retention gap; the latter was connected to a manually operated split valve. As previously demonstrated, the use of a split valve enables the regulation of gas flows through both analytical columns, generating the most appropriate gas linear velocities. Comprehensive 2D GC experiments were carried out on Arabica roasted coffee volatiles (previously extracted by means of solid-phase microextraction) with the split-valve closed (equal to what can be defined as conventional GC x GC) and with the split-valve opened at various degrees. The reasons why it is absolutely not effective to use a 0.05 mm ID column as second dimension in a conventional GC x GC instrument will be discussed and demonstrated. On the contrary, the use of a 0.05 mm ID column as second dimension, under ideal conditions in a split-flow, twin-oven system, will also be illustrated and discussed.

Rapid SPE-HPLC determination of the 16 European priority polycyclic aromatic hydrocarbons in olive oils.

Polycyclic aromatic hydrocarbons (PAHs) are a large class of organic compounds. It has been established that the main source of exposure to these compounds for human beings is through food, particularly fats and oils, due to the lipophilic nature of these polycyclic compounds. The aim of this work was to optimise and validate a method involving SPE and HPLC for rapid determination of the 16 European Union (EU) priority PAHs (required by the Recommendation 2005/108/EC) in vegetable oils. Two spectrofluorometric detectors and a UV-Visible detector in series were used to identify and quantify the target compounds. Linearity, recoveries, LOD, and LOQ were found to be in agreement with the performance criteria for benzo[a]pyrene (BaP) analysis as required by the Commission Directive 2005/10/EC, and satisfactory for all the compounds of interest, except for cyclopenta[c,d]pyrene, which presented a very low signal in the UV. Optimised chromatographic conditions for the separation of 25 PAHs, comprising both EPA and EU priority PAHs plus benzo[e]pyrene and benzo[b]chrysene, have been also proposed.

Rapid validated method for the analysis of benzo[a]pyrene in vegetable oils by using solid-phase microextraction-gas chromatography-mass spectrometry.

A solid-phase microextraction method coupled with comprehensive gas chromatography and time-of-flight mass spectrometry for the determination of polycyclic aromatic hydrocarbons in vegetable oils has recently been reported. The present paper tested the possibility to use the solid-phase microextraction method coupled with one-dimensional gas chromatography-mass spectrometry of the only benzo[a]pyrene. Furthermore, an in-house validation for benzo[a]pyrene, used as a marker, as requested by the European regulation no. 208/2005, was carried out. Statistical tests were performed to elaborate the data. Linearity was satisfactory (r(2)=0.999), between about 0.5 and 15 microg/kg. Detection limit and quantification limit were 0.17 and 0.46 microg/kg, respectively. In-day and inter-day repeatability were less than 6% in both cases.

Determination of polycyclic aromatic hydrocarbons in vegetable oils using solid-phase microextraction-comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry.

A simple and fast solid-phase microextraction method coupled with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry was developed for analysis of polycyclic aromatic hydrocarbons in edible oil, performed directly in a hexane solution of the oil. Sampling conditions (solvent used, extraction time, extraction temperature and fiber rinsing time) were optimized by using a sample of oil fortified with a standard solution of polycyclic aromatic hydrocarbons. The method was validated by calculating linear range, correlation coefficient, accuracy, repeatability, detection limit and quantification limit. The method was applied to several oils collected from the market and directly from an olive pomace extraction plant.