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.

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.

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.

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.

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.

Simultaneous determination of volatile and semi-volatile aromatic hydrocarbons in virgin olive oil by headspace solid-phase microextraction coupled to gas chromatography/mass spectrometry.

A reliable, simple and relatively fast method for the simultaneous determination of volatile and semi-volatile aromatic hydrocarbons in virgin olive oil was developed, based on headspace solid-phase microextraction (HS-SPME). The investigation regarded eco-contaminants such as alkylated monoaromatic hydrocarbons from C1- to C4-benzenes and light polyaromatic hydrocarbons up to four aromatic rings. Sampling and chromatographic conditions were optimized by using standard solutions in deodorized olive oil and the analytical performances of the method were determined. The proposed method was then applied to real samples of virgin olive oil were the target hydrocarbons could be identified and quantified. Several of them had not been previously quantified in virgin olive oil. Moreover, by the analysis of olive oil samples an additional number of C4-benzenes could be tentatively identified.

HPLC determination of free nitrogenous compounds of Centaurea solstitialis (Asteraceae), the cause of equine nigropallidal encephalomalacia.

Centaurea solstitialis (yellow star thistle) has been proven to cause equine nigropallidal encephalomalacia in horses. Over the last fifty years, nigropallidal encephalomalacia has been of interest to human medicine due to the possible connection with Parkinson's disease. Previous studies indicated the presence of neurotoxic nitrogenous compounds in polar extracts of the plant. In order to give a more detailed description of the nitrogen-containing fraction of C. solstitialis, various samples were collected at different development stages. Different aliquots of the same aqueous extract were directly derivatized with o-phthaldialdehyde and dansyl chloride and analyzed separately by reversed-phase HPLC. A complete profile of the free nitrogenous fraction of C. solstitialis was given and results obtained with the two derivatization procedures were compared. No particularly high level of free aspartic and glutamic acids, two potent neuroexcitotoxic amino acids, were found in polar extracts of the plant. Tyramine resulted to be the most important biologically active amine present in C. solstitialis (with a mean concentration of 2.0 mg/100 g of dry weight).

Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: modifications induced by oxidation and suitable markers of oxidative status.

Modifications of virgin olive oil subjected to accelerated storage were evaluated by HS-SPME analysis. To find a suitable marker of oxidative degradation, the volatile compounds showing variable concentration during the oxidative process have been identified and quantified by SPME coupled to GC-MS and GC-FID, respectively. The SPME analysis results were then compared with the parameters usually applied to assess the oxidative status of lipids, such as peroxide value, spectrophotometric absorbance, and loss of unsaturated fatty acids. Finally, the assessment of nonanal has been suggested as a marker of oxidative degradation. This rapid, inexpensive, and reliable method may allow screening of oils prior to testing by a panel of assessors.

Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: characterization of virgin olive oils from two distinct geographical areas of northern Italy.

SPME was employed to characterize the volatile profile of virgin olive oils produced in two geographical areas of northern Italy: the region of the Gulf of Trieste and the area near Lake Garda. There are as yet no data on the headspace composition of virgin olive oils from these regions, characterized by particular conditions of growth for Olea europaea. Using the SPME technique coupled to GC-MS and GC-FID, the volatile components of 42 industrially produced virgin olive oil samples were identified and the principal compounds quantitatively analyzed. Significant differences in the proportion of volatile constituents from oils of different varieties and geographical origins were detected. The results suggest that besides the genetic factor, environmental conditions influence the volatile formation.

Presence of phytosterol oxides in crude vegetable oils and their fate during refining.

The content of phytosterol oxidation products was determined in samples of crude vegetable oils: peanut, sunflower, maize, palm nut, and lampante olive oils that were intended for refining and not for direct consumption. The 7 alpha- and 7 beta-hydroxy derivatives of beta-sitosterol, stigmasterol, and campesterol and the 7-keto-beta-sitosterol were the principal phytosterol oxides found in almost all of the oils analyzed. In some oils, the epoxy and dihydroxy derivatives of beta-sitosterol were also found at very low levels. The highest total concentrations of phytosterol oxides, ranging from 4.5 to 67.5 and from 4.1 to 60.1 ppm, were found in sunflower and maize oils, respectively. Lower concentrations were present in the peanut oils, 2.7-9.6 ppm, and in the palm nut oil, 5.5 ppm, whereas in the lampante olive oils, only three samples of the six analyzed contained a low concentration (1.5-2.5 ppm) of oxyphytosterols. No detectable levels of phytosterol oxides were found in the samples of palm and coconut oils. Bleaching experiments were carried out on a sample of sunflower oil at 80 degrees C for 1 h with 1 and 2% of both acidic and neutral earths. The bleaching caused a reduction of the hydroxyphytosterol with partial formation of steroidal hydrocarbons with three double bonds in the ring system at the 2-, 4-, and 6-positions (steratrienes). The same sunflower oil was deodorized at 180 degrees C under vacuum for 1 h, and no dehydration products were formed with a complete recovery of the hydroxyphytosterols. A bleaching test with acidic earths was carried out also with an extra virgin olive oil fortified with 7-keto-cholesterol, dihydroxycholesterol, and alpha-epoxy-cholesterol. There was no formation of steratrienes from these compounds, but dihydroxycholesterol underwent considerable decomposition and alpha-epoxycholesterol underwent ring opening with formation of the dihydroxy derivative, whereas 7-ketocholesterol was rather stable

Analysis of virgin olive oil volatile compounds by headspace solid-phase microextraction coupled to gas chromatography with mass spectrometric and flame ionization detection.

The efficiency of headspace solid-phase microextraction (SPME) was evaluated for the qualitative and semi-quantitative analysis of virgin olive oil volatile compounds. The behaviour of four fibre coatings was compared for sensitivity, repeatability and linearity of response. A divinylbenzene-Carboxen-polydimethylsiloxane fibre coating was found to be the most suitable for the analysis of virgin olive oil volatiles. Sampling and chromatographic conditions were examined and the SPME method, coupled to GC with MS and flame ionization detection, was applied to virgin olive oil samples. More than 100 compounds were isolated and characterised. The presence of some of these compounds in virgin olive oil has not previously been reported. The main volatile compounds present in the oil samples were determined quantitatively.