Endogenous n-Alkanes in Vegetable Oils: Validation of a Rapid Offline SPE-GC-FID Method, Comparison with Online LC-GC-FID and Potential for Olive Oil Quality Control.

The potential of endogenous n-alkane profiling for the assessment of extra virgin olive oils (EVOO) adulteration (blends with cheaper vegetable oils) has been studied by relatively few authors. Analytical methods used for this purpose often involve tedious and solvent-intensive sample preparation prior to analytical determination, making them unattractive. A rapid and solvent-sparing offline solid phase extraction (SPE) gas chromatography (GC) flame ionization detection (FID) method for the determination of endogenous n-alkanes in vegetable oils was, therefore, optimized and validated. The optimized method demonstrated good performance characteristics in terms of linearity (R2 > 0.999), recovery (on average 94%), and repeatability (residual standard deviation, RSD < 11.9%). The results were comparable to those obtained with online high-performance liquid chromatography (HPLC)-GC- FID ( RSD < 5.1%). As an example of an application to prove the potentiality of endogenous n-alkanes in revealing frauds, the data set obtained from 16 EVOO, 9 avocado oils (AVO), and 13 sunflower oils (SFO), purchased from the market, was subjected to statistical analysis and principal component analysis. Two powerful indices, namely (n-C29 + n-C31)/(n-C25 + n-C26) and n-C29/n-C25, were found to reveal the addition of 2% SFO in EVOO and 5% AVO in EVOO, respectively. Further studies are needed to confirm the validity of these promising indices.

Pressurized Liquid Extraction: A Powerful Tool to Implement Extraction and Purification of Food Contaminants.

Pressurized liquid extraction (PLE) is considered an advanced extraction technique developed in the mid-1990s with the aim of saving time and reducing solvent with respect to traditional extraction processes. It is commonly used with solid and semi-solid samples and employs solvent extraction at elevated temperatures and pressures, always below the respective critical points, to maintain the solvent in a liquid state throughout the extraction procedure. The use of these particular pressure and temperature conditions changes the physicochemical properties of the extraction solvent, allowing easier and deeper penetration into the matrix to be extracted. Furthermore, the possibility to combine the extraction and clean-up steps by including a layer of an adsorbent retaining interfering compounds directly in the PLE extraction cells makes this technique extremely versatile and selective. After providing a background on the PLE technique and parameters to be optimized, the present review focuses on recent applications (published in the past 10 years) in the field of food contaminants. In particular, applications related to the extraction of environmental and processing contaminants, pesticides, residues of veterinary drugs, mycotoxins, parabens, ethyl carbamate, and fatty acid esters of 3-monochloro-1,2-propanediol and 2-monochloro-1,3-propanediol from different food matrices were considered.

The Determination of Triacylglycerols and Tocopherols Using UHPLC-CAD/FLD Methods for Assessing the Authenticity of Coffee Beans.

The authenticity of coffee beans was addressed in this study using an analytical method with minimal sample preparation to achieve simple oil extraction and through the implementation of cost-effective equipment. For this purpose, methods using UHPLC with CAD and FLD detectors were applied to detect triglycerides and tocopherols in coffee, respectively. The coffee samples included two main varieties: Arabica from Brazil, Colombia, Ethiopia, and Uganda, as well as the Robusta variety from Cambodia, Guatemala, India, and Vietnam. The samples were either in their green state or subjected to different roasting levels. The used methods successfully distinguished the Arabica and Robusta variants targeted in this study based on their tocopherols and TAG profiles, with the latter being particularly effective for discriminating the origins of the Arabica coffee, while tocopherols excelled at differentiating the origin of the Robusta coffee. TAGs and tocopherols were not affected by the type of roasting, from medium to very dark, suggesting it is possible to distinguish between coffee varieties independently from their degree of roasting. The obtained results hold valuable implications for future research regarding coffee fraud and authenticity.

Monitoring and Occurrence of Heavy PAHs in Pomace Oil Supply Chain Using a Double-Step Solid-Phase Purification and HPLC-FLD Determination.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental and processing contaminants generated by both spontaneous and anthropogenic incomplete combustion processes of organic matter. Contamination of PAHs in vegetable oils can result from several factors and processes, including environmental contamination, oil processing, and migration from food contact materials. The determination of PAHs in edible oil presents a challenge because of the complexity of the matrix. Since PAHs are present at lower levels than triglycerides, it is necessary to isolate the compounds of interest from the rest of the matrix. To this purpose, a new purification approach based on a double solid-phase extraction (SPE) step followed by high performance liquid chromatography-fluorometric detector (HPLC-FLD) analysis was developed. The method involves a first purification step by using a 5 g silica SPE cartridge, previously washed with dichloromethane (20 mL), dried completely, and then conditioned with n-hexane (20 mL). The triglycerides are retained by the silica, while the PAH-containing fraction is eluted with a mixture of n-hexane/dichloromethane (70/30, v/v). After evaporation, the residue is loaded on a 5 g amino SPE cartridge and eluted with n-hexane/toluene (70/30, v/v) before HPLC-FLD analysis. The focus was the evaluation of the contribution of the various phases of the pomace oil supply chain in terms of the heavy PAHs (PAH8) concentration. Data collected showed that pomace contamination increased (by 15 times) as storage time increased. In addition, the process of pomace drying, which is necessary to reduce its moisture content before solvent extraction of the residual oil, appeared to significantly contribute to the total heavy PAHs content, with increases in value by up to 75 times.

Evidence of the Possible Interaction between Ultrasound and Thiol Precursors.

The effect of ultrasound (20 kHz, 153 µm) on the prefermentation extraction mechanisms in Sauvignon Blanc grapes was studied, focusing on 3-mercaptohexan-1-ol (3MH) and 4-mercapto-4-methyl-pentan-2-one (4MMP) precursors linked to glutathione (GSH) and cysteine (Cys). The treatment determined a positive extraction trend between the duration (untreated, 3 and 5 min) and the conductivity or the concentration of catechins and total phenols, significantly differentiated after 5 min. Nevertheless, the concentration of the thiol precursors in grape juice not only remained undifferentiated, but that of 3-S-glutathionyl mercaptohexan-1-ol showed a negative trend with the treatment time applied (168 ± 43, 156 ± 36, and 149 ± 32 µg/L, respectively, for control, 3 and 5 min). The divergence on the effect between families of compounds suggests an interaction between the sonication treatment and thiol precursor molecules. In order to evaluate the possible degradation properly, ultrasound was applied in a model solution spiked with 3MH and 4MMP precursors, reproducing the conditions of grapes. Except for Cys-3MH, the mean concentration (n = 5) for the rest of the precursors was significantly lower in treated samples, predominantly in those linked to glutathione (~-22% and ~18% for GSH-3MH and GSH-4MMP) rather than to cysteine (~-6%~-8% for Cys-3MH and Cys-4MMP). The degradation of precursors was associated with a significant increase of 3MH and 4MMP. The formation of volatile thiols following sonication is interesting from a technological point of view, as they are key aroma compounds of wine and potentially exploitable in the wine industry through specific vinification protocols.

In-pipette solid-phase extraction prior to flow-modulation comprehensive two-dimensional gas chromatography with dual detection for the determination of minor components in vegetable oils.

The present research is based on the development of an effective, environmentally-friendly and low-cost method for investigation of minor components in vegetable oils, exploiting the advantages of a miniaturized solid-phase extraction (SPE) and the potential of flow modulation (FM) comprehensive two-dimensional gas chromatography (GC×GC), coupled to a mass spectrometer (MS) and a flame ionization detector (FID). The initial sample preparation step was carried out using a miniaturized approach characterized by a SPE process in a Pasteur pipette. Then, the isolated fraction was injected into an FM GC×GC system. Tentative identification was carried out by means of MS spectral information, while quantification was carried out by using the FID data. Four different vegetable oil samples were analyzed using the proposed method, namely an extra virgin olive oil (EVO), a hazelnut oil (HO), a borage oil, and an EVO/HO mixture.

Mineral oil saturated hydrocarbons (MOSH) in female Fischer 344 rats; accumulation of wax components; implications for risk assessment.

Female Fischer 344 rats were exposed to three MOSH mixtures: oils largely below and above C25 (S-C25 and L-C25) and a 1:1 mixture of L-C25 with a wax; doses of 400, 1000 and 4000mg/kg feed were administered during 120days. MOSH were determined by on-line HPLC-GC-FID in liver, spleen, adipose tissue and the carcass. The composition of the hydrocarbons accumulated in the tissues was further analyzed by comprehensive two-dimensional GC (GC×GC). MOSH in the mass range of C26-30 were more strongly accumulated than those between C20-25, which does not support the present classification of MOSH differentiating at n-C25 for risk assessment. Compared to the total of the MOSH, n-alkanes and n-alkyl monocyclic naphthenes were generally enriched in adipose tissue. In liver and spleen, n-alkanes up to C25 were eliminated, but strongly accumulated at around C30. Based on this profile, poor solubility and the melting points, it is hypothesized that crystallization protects these wax components against metabolism and elimination. In the liver, relative retention of n-alkanes decreased again beyond C30, accentuated at high exposure, suggesting reduced absorption. Compared to the animal data, accumulation of n-alkanes from food sources, such as apples, into human tissues seems low, perhaps because of low absorption due to their presence in crystalline form. A series of dominant isoalkanes, accumulated in all tissues analyzed, was characterized, though without proposing a structure. Implications on present regulation of white mineral oil products are discussed.

Accumulation of mineral oil saturated hydrocarbons (MOSH) in female Fischer 344 rats: Comparison with human data and consequences for risk assessment.

Female Fischer 344 rats were orally exposed to a mixture of mineral oil saturated hydrocarbons (MOSH) of broad molecular mass range at doses of 40, 400 and 4000mg/kg feed. Amounts and compositions of the MOSH were analyzed in liver, spleen, adipose tissue and the carcass after exposure during 30, 60, 90 and 120d as well as after 90d exposure followed by 30d depuration. At 40mg/kg in the feed, after 30d of exposure, 10.9% of the ingested MOSH were recovered from the animal body; after 90d plus 30d depuration it was 3.9%. In liver and spleen, the maximum retention in terms of molecular mass (simulated distillation) was at n-C29; in adipose tissue and carcass it was at n-C15/16. The differentiation between MOSH below and above n-C25 (Class I versus Class II and III oils), used for present regulation, is not supported by the present data on accumulation; structural characteristics seem more pertinent than molecular mass. Concentrations in the tissues increased far less than proportionally with the dose, rendering linear extrapolation to low doses questionable. No steady state was reached after 120d. In fact, comparing with the concentrations in human tissues at the estimated exposure, extrapolation from animal experiments seems to grossly underestimate human internal exposure. Comprehensive two-dimensional gas chromatography (GCxGC) was used to characterize the MOSH residues in the tissues with the aim of identifying the most strongly accumulated types. In the liver and spleen, the highly branched hydrocarbons dominated, whereas in the adipose tissue it was the n-alkanes and species with main n-alkyl moieties. Strong MOSH accumulation is not of concern per se, but the safety at the high concentrations in human tissues needs to be re-evaluated, possibly taking into account also end points other than granuloma formation.

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.

Microwave assisted saponification (MAS) followed by on-line liquid chromatography (LC)-gas chromatography (GC) for high-throughput and high-sensitivity determination of mineral oil in different cereal-based foodstuffs.

A high throughput, high-sensitivity procedure, involving simultaneous microwave-assisted extraction (MAS) and unsaponifiable extraction, followed by on-line liquid chromatography (LC)-gas chromatography (GC), has been optimised for rapid and efficient extraction and analytical determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) in cereal-based products of different composition. MAS has the advantage of eliminating fat before LC-GC analysis, allowing an increase in the amount of sample extract injected, and hence in sensitivity. The proposed method gave practically quantitative recoveries and good repeatability. Among the different cereal-based products analysed (dry semolina and egg pasta, bread, biscuits, and cakes), egg pasta packed in direct contact with recycled paperboard had on average the highest total MOSH level (15.9 mg kg(-1)), followed by cakes (10.4 mg kg(-1)) and bread (7.5 mg kg(-1)). About 50% of the pasta and bread samples and 20% of the biscuits and cake samples had detectable MOAH amounts. The highest concentrations were found in an egg pasta in direct contact with recycled paperboard (3.6 mg kg(-1)) and in a milk bread (3.6 mg kg(-1)).

Improvement of mineral oil saturated and aromatic hydrocarbons determination in edible oil by liquid-liquid-gas chromatography with dual detection.

Mineral oils, which are mainly composed of saturated hydrocarbons and aromatic hydrocarbons, are widespread food contaminants. Liquid chromatography coupled to gas chromatography with flame ionization detection represents the method of choice to determine these two families. However, despite the high selectivity of this technique, the presence of olefins (particularly squalene and its isomers) in some samples as in olive oils, does not allow the correct quantification of the mineral oil aromatic hydrocarbons fraction, requiring additional off-line tools to eliminate them. In the present research, a novel on-line liquid chromatography coupled to gas chromatography method is described for the determination of hydrocarbon contamination in edible oils. Two different liquid chromatography columns, namely a silica one (to retain the bulk of the matrix) and a silver-ion one (which better retains the olefins), were coupled in series to obtain the mineral oil aromatic hydrocarbons hump free of interfering peaks. Furthermore, the use of a simultaneous dual detection, flame ionization detector and triple quadrupole mass spectrometer allowed us not only to quantify the mineral oil contamination, but also to evaluate the presence of specific markers (i.e. hopanes) to confirm the petrogenic origin of the contamination.

Determination of phthalate esters in vegetable oils using direct immersion solid-phase microextraction and fast gas chromatography coupled with triple quadrupole mass spectrometry.

Phthalates are a group of synthetic compounds mainly used as plasticizers, which have been classified as endocrine-disrupting chemicals and potential human-cancer causing agents. They can be found in high amounts in foods, deriving mainly from plastic packaging. The analytical determination of these compounds is very challenging since they are ubiquitous. Therefore, minimization of sample manipulation is highly desirable. The present work exploited the application of a solid-phase microextraction method for the analysis of phthalates in vegetable oil. A preliminary comparison between a polydimethylsiloxane (PDMS) and a Carbopack Z/PDMS fiber was carried out both in the headspace and direct immersion extraction modes. Before immersing the fiber, a rapid liquid-liquid extraction was performed using acetonitrile to remove the bulk of triglycerides. PDMS in the direct immersion mode showed the best performance. The method was fully validated obtaining a good linearity with a coefficient of correlation of over 0.9960 for all compounds, repeatability and accuracy values generally better than 10%, and very good limit of quantification values.

Migration of selected hydrocarbon contaminants into dry semolina and egg pasta packed in direct contact with virgin paperboard and polypropylene film.

Migration of mineral oil saturated hydrocarbons (MOSH), polyolefin oligomeric saturated hydrocarbons (POSH), and polyalphaolefins (PAO from hot melts) into dry semolina and egg pasta packed in direct contact with virgin paperboard or polypropylene (PP) flexible film was studied. Migration was monitored during shelf life (up to 24 months), through storage in a real supermarket (packs kept on shelves), conditions preventing exchange with the surrounding environment (packs wrapped in aluminium foil), and storage in a warehouse (packs inside of the transport box of corrugated board). Semolina pasta packed in virgin paperboard (without hot melts) had a MOSH content lower than 1.0 mg kg(-1). An increasing contamination with PAO belonging to the adhesives used to close the boxes was detected in egg pasta, wrapped in aluminium (1.5 and 5 mg kg(-1) after 3 and 24 months, respectively). An environmental contribution to total hydrocarbon contamination was observed in egg pasta kept on shelves that, after 3 and 24 months, showed levels of PAO/MOSH < C25 around 3 and 10 mg kg(-1), respectively. The migration of POSH from PP film into egg pasta wrapped in aluminium was around 0.6 mg kg(-1) after 3 months of contact and reached 1.7 mg kg(-1) after 24 months of contact. After 9 months of contact, semolina pasta packed in PP film and stored in the transport box showed that some MOSH migrated into the pasta from the board of the transport box (through the plastic film).

Comparison of different injection modes in edible oil minor components analysis.

Waxes and fatty acid alkyl esters are minor components used as official parameters to control the authenticity and quality of a high-value olive oil product. A poor measurement can lead to a misleading classification of the oil. The official method requires their analysis together by capillary gas chromatography equipped with a flame ionization detector and an on-column injector to avoid discrimination and thermal degradation. The degradation can occur to a different extent if different (and not properly optimized) injectors are used. However, other injection techniques, such as programmed-temperature vaporizer, are much more versatile and more widespread. The aim of the present work was to compare the performance of a programmed-temperature vaporizer injector, in on-column and splitless mode, with the on-column injector to analyze alkyl esters and waxes. Discrimination among high-boiling compounds was evaluated, as well as the occurrence of thermal degradation, especially of sterols and diterpene alcohol (phytyl and geranylgeraniol) esters. A proper optimization of a programmed-temperature vaporizer injection, with particular attention to the liner selection, was proven to provide comparable results to the traditional on-column injection. A performance comparison was carried out both on standard mixtures and on real oil samples.

Migration of selected hydrocarbon contaminants into dry pasta packaged in direct contact with recycled paperboard.

This paper deals with the migration of selected hydrocarbon contaminants, namely mineral oil hydrocarbons (MOH), diisopropyl naphthalenes (DIPN) and polyalphaolefins (PAO) from adhesives into dry semolina and egg pasta packaged in direct contact with recycled paperboard. Migration was monitored during its shelf life (for up to two years) simulating storage in a supermarket (packs on shelves) and conditions preventing exchange with the surrounding environment (packs wrapped in aluminium foil). Migration from the secondary packaging (transport boxes of corrugated board) was also studied for semolina pasta. After 24 months of exposure, semolina pasta stored on shelves reached 3.2 and 0.6 mg kg(-1) of MOSH and MOAH, respectively, Migration from the adhesives used to close the boxes and from the transport boxes contributed about 30% and 25% of the total contamination, respectively. The highest contamination levels (14.5 and 2.0 mg kg(-1) of MOSH and MOAH, respectively, after 24 months) were found in egg pasta stored on shelves (no adhesives), and seemed due to the highest contribution from the external environment.

Mineral oil in human tissues, part II: characterization of the accumulated hydrocarbons by comprehensive two-dimensional gas chromatography.

Mineral oil hydrocarbons are by far the largest contaminant in the human body. Their composition differs from that in the mineral oils humans are exposed to, and varies also between different tissues of the same individual. Using the presently best technique for characterizing the composition of mineral oil hydrocarbons, comprehensive two-dimensional gas chromatography (GC×GC), the hydrocarbons in human tissues were compared to those of various mineral oils. This provided information about the strongly accumulated species and might give hints on the flow path through the human body. The selectivity of accumulation is probably also of interest for the risk assessment of synthetic hydrocarbons (polyolefins). GC×GC grouped the MOSH into classes of n-alkanes, paraffins with a low degree of branching, multibranched paraffins and naphthenes (alkylated cyclic hydrocarbons) with 1-4 rings. Metabolic elimination was observed for constituents of all these classes, but was selective within each class. The MOSH in the subcutaneous abdominal fat tissues and the mesenteric lymph nodes (MLN) had almost the same composition and included the distinct signals observed in mineral oil, though in reduced amounts relative to the cloud of unresolved hydrocarbons. The MOSH in the liver and the spleen were different from those in the MLN and fat tissue, but again with largely identical composition for a given individual. Virtually all constituents forming distinct signals were eliminated, leaving an unresolved residue of highly isomerized hydrocarbons.

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.

Mineral oil in human tissues, Part I: concentrations and molecular mass distributions.

Of 37 subjects aged 25-91 y (mean 67 y), mineral oil hydrocarbons were measured in subcutaneous abdominal fat tissue, mesenteric lymph nodes (MLN), spleen, liver and lung, for some of them also in kidney, heart and brain. No mineral oil aromatic hydrocarbons (MOAH) were detected. The mean concentration of mineral oil saturated hydrocarbons (MOSH) in the mesenteric lymph nodes was 223 mg/kg, in liver 131 mg/kg, in fat tissue 130 mg/kg, in spleen 93 mg/kg and in lung 12 mg/kg. They were clearly lower in kidney, heart and brain. The maxima, found in MLN and spleen, were 1390 and 1400 mg/kg, respectively. For a quarter of the subjects a total amount of MOSH in the body above 5 g was calculated. The MOSH composition in the fat tissue and the MLN appeared virtually identical and varied little between the subjects. It was centered on the n-alkanes C23-C24, ranged from C16 to C35 and included hydrocarbons of plant origin. The MOSH in spleen and liver had almost the same composition for a given subject, but varied somewhat between subjects. They were centered between C25 and C27, ranged from C18 to beyond C45 and were without hydrocarbons of plant origin. Part of the MOSH seem to be strongly accumulated, resulting in far higher concentrations in human tissues related to exposure than observed in shorter term animal experiments. The composition of the accumulated MOSH does not support that Class I mineral oils, sometimes termed "food grade", are less accumulated in the human body than Class II and III oils, which questions the present classification.

Optimisation of pressurised liquid extraction (PLE) for rapid and efficient extraction of superficial and total mineral oil contamination from dry foods.

Pressurised liquid extraction (PLE) represents a powerful technique which can be conveniently used for rapid extraction of mineral oil saturated (MOSH) and aromatic hydrocarbons (MOAH) from dry foods with a low fat content, such as semolina pasta, rice, and other cereals. Two different PLE methods, one for rapid determination of superficial contamination mainly from the packaging, the other for efficient extraction of total contamination from different sources, have been developed and optimised. The two methods presented good performance characteristics in terms of repeatability (relative standard deviation lower than 5%) and recoveries (higher than 95%). To show their potentiality, the two methods have been applied in combination on semolina pasta and rice packaged in direct contact with recycled cardboard. In the case of semolina pasta it was possible to discriminate between superficial contamination coming from the packaging, and pre-existing contamination (firmly enclosed into the matrix).

Optimization of pressurized liquid extraction (PLE) for rapid determination of mineral oil saturated (MOSH) and aromatic hydrocarbons (MOAH) in cardboard and paper intended for food contact.

Packaging can represent a primary source of food contamination with mineral oil saturated hydrocarbons (MOSH) and aromatic hydrocarbons (MOAH), especially when recycled cardboard or mineral oil based printing inks are used. A pressurized liquid extraction (PLE) method, followed by on-line LC-GC analysis, has been optimized for rapid mineral oil determination in cardboard and paper samples. The proposed method involves extraction with hexane (2 cycles) at 60°C for 5 min, and allows for the processing of up to 6 samples in parallel with minimal sample manipulation and solvent consumption. It gave good repeatability (coefficient of variation lower than 5%) and practically quantitative extraction yield (less than 2% of the total contamination found in a third separate cycle). The method was applied to different cardboards and paper materials intended for food contact. Results obtained were similar to those obtained by applying classical solvent extraction with hexane/ethanol 1:1 (v/v) as described by Lorenzini et al. [20].