Food lipid oxidation under gastrointestinal digestion conditions: A review.

Unravelling the relationship between food and health requires a more in-depth knowledge of the various changes occurring in the gastrointestinal tract during digestion and which may ultimately affect the nutritional quality and safety of ingested food lipids before absorption into the bloodstream. In this context, this review deals with the oxidation process of food lipids under digestive conditions and the studies carried out on this topic using different digestion models: in vitro, in vivo or ex vivo, static or dynamic, and including one, two and/or three digestive phases (oral, gastric and duodenal). These studies have contributed to clarifying the occurrence and extent of lipid degradation under such a particular environment, many of them also highlighting the factors affecting the advance or delay of the oxidation of dietary lipids during digestion, like: food lipid content, unsaturation degree and initial oxidative status; the presence in the food bolus of compounds showing antioxidant activity (polyphenols, tocopherols…) either added or naturally present; the presence in the food bolus of proteins (including iron or not); food technological or culinary processings (salting, smoking, cooking…), among others. Likewise, the methodologies employed to study lipid oxidation under digestive conditions are also summarized and future research perspectives are discussed.

Influence of different salting processes on the evolution of the volatile metabolites of vacuum-packed fillets of farmed and wild sea bass (Dicentrarchus labrax) stored under refrigeration conditions: a study by SPME-GC/MS.

BACKGROUND: Fish shelf-life extension is a topic of great interest. In this study the behaviour of salted and unsalted farmed and wild European sea bass (Dicentrarchus labrax) fillets during storage was analysed through the evolution of their volatile metabolites. Farmed and wild sea bass fillets were brine-salted for 15 or 75 min, or dry-salted, vacuum-packed and stored at 4 °C for up to 1 month, and their headspaces were studied by Solid Phase Micro extraction-Gas Chromatography/Mass Spectrometry (SPME-GC/MS). RESULTS: At the same storage time, unsalted wild fillets contained, in general, a higher number and abundance of volatile compounds coming from microbiological or endogenous enzymatic activity than unsalted farmed ones. The more intense the salting, the lower the number and abundance of microbiological spoilage metabolites, especially in wild samples. The appearance of oxidation metabolites only in dry-salted wild samples evidences that this kind of salting provokes a certain oxidation in these samples. CONCLUSIONS: The better performance of farmed than wild fillets suggests that salted farmed fillets, vacuum-packed and stored under refrigeration conditions, could be a successful alternative to diversify the presence of sea bass in the market. © 2016 Society of Chemical Industry.

Farmed and wild sea bass (Dicentrarchus labrax) volatile metabolites: a comparative study by SPME-GC/MS.

BACKGROUND: Farmed and wild European sea bass (Dicentrarchus labrax) could be distinguished by its volatile metabolites, an issue not addressed until now. The aim of this work was to study these metabolites by solid-phase microextraction followed by gas chromatography/mass spectrometry (SPME-GC/MS). RESULTS: Both farmed and wild sea bass have a great number of volatile metabolites, most of them being in low concentrations. These include alcohols, aldehydes, ketones, alkylfurans, acids, aliphatic and aromatic hydrocarbons, terpenes, sulfur and nitrogen derivatives, 2,6-di-tert-butyl-4-methylphenol and one derived compound, as well as 2,4,7,9-tetramethyl-5-decyne-4,7-diol, this latter compound presumably resulting from environmental contamination. Important differences have been detected between both types of sea bass, and also among individuals inside each group. Farmed specimens are richer in volatile metabolites than the wild counterparts; however, these latter, in general, contain a high number and abundance of metabolites resulting from microbial and enzymatic non-oxidative activity than the former. CONCLUSIONS: Clear differences in the volatile metabolites of wild and farmed sea bass have been found. A great deal of valuable information on sea bass volatile metabolites has been obtained, which can be useful in understanding certain aspects of the quality and safety of raw and processed sea bass.

2,6-Di-Tert-Butyl-Hydroxytoluene and Its Metabolites in Foods.

2,6-Di-tert-butyl-hydroxytoluene (BHT, E-321) is a synthetic phenolic antioxidant which has been widely used as an additive in the food, cosmetic, and plastic industries for the last 70 y. Although it is considered safe for human health at authorized levels, its ubiquitous presence and the controversial toxicological data reported are of great concern for consumers. In recent years, special attention has been paid to these 14 metabolites or degradation products: BHT-CH2 OH, BHT-CHO, BHT-COOH, BHT-Q, BHT-QM, DBP, BHT-OH, BHT-OOH, TBP, BHQ, BHT-OH(t), BHT-OH(t)QM, 2-BHT, and 2-BHT-QM. These derived compounds could pose a human health risk from a food safety point of view, but they have been little studied. In this context, this review deals with the occurrence, origin, and fate of BHT in foodstuffs, its biotransformation into metabolites, their toxicological implications, their antioxidant and prooxidant properties, the analytical determination of metabolites in foods, and human dietary exposure. Moreover, noncontrolled additional sources of exposure to BHT and its metabolites are highlighted. These include their carryover from feed to fish, poultry and eggs, their presence in smoke flavorings, their migration from plastic pipelines and packaging to water and food, and their presence in natural environments, from which they can reach the food chain.

Fourier transform infrared spectroscopy as a tool to study farmed and wild sea bass lipid composition.

BACKGROUND: The lipids of 16 farmed and wild European sea bass (Dicentrarchus labrax) samples were studied by Fourier transform infrared (FTIR) spectroscopy. The spectroscopic parameters which would be useful when distinguishing between both fish origins were analysed. RESULTS: It was shown, for the first time, that the frequency and the ratio between the absorbance of certain bands are efficient and reliable authentication tools for the origin of sea bass. Furthermore, relationships between infrared data and fish lipids composition referring to the molar percentage or concentration of certain acyl groups were also studied. It was proved that some infrared spectroscopic data (the frequency of certain bands or the ratio of the absorbance of others), are very closely related to the composition of sea bass lipids. It was shown for the first time that certain infrared spectroscopic data could predict, with a certain degree of approximation, the molar percentage, or concentration, of omega-3, docosahexaenoic (DHA) and di-unsaturated omega-6 (linoleic) in sea bass lipids. CONCLUSION: The consistency of the results confirms the usefulness of FTIR spectroscopy to detect frauds regarding sea bass origin, and to provide important compositional data about sea bass lipids from the nutritional and technological point of view.

Volatile components of several virgin and refined oils differing in their botanical origin.

BACKGROUND: The volatile components of virgin and refined oils whose compositions were very different in terms of oleic, linoleic and linolenic acyl groups were studied by solid phase microextraction (SPME) followed by gas chromatography/mass spectrometry (GC/MS), and their composition of acyl groups was determined by proton nuclear magnetic resonance (1) H NMR. The virgin oils studied were olive, linseed and walnut oils, while the refined oils were sunflower and rapeseed oils. RESULTS: Large differences were found among the headspace compositions of the virgin oils. These were due in part to compounds that could be considered as markers of different vegetable origins and manufacturing processes, but also to compounds resulting from degradative oxidation of the main acyl group components. By virtue of their being refined, fewer components were present in the refined oils, although differences in their headspaces were found owing to the presence of compounds resulting from oxidation of the determined acyl groups. CONCLUSION: The studied oils show specific characteristics regarding not only their main components, which are well known, but also their volatile components. The latter provide information on some of the processes to which oils have been submitted, on their oxidation level, on their botanical origin and on the compounds responsible for their odour.

Different Effects of Vitamin C-Based Supplements on the Advance of Linseed Oil Component Oxidation and Lipolysis during In Vitro Gastrointestinal Digestion.

Although widely consumed, dietary supplements based on Vitamin C contain high doses of this compound, whose impact on lipid oxidation during digestion needs to be addressed. Therefore, the effect of seven commercial supplements and of pure l-ascorbic acid and ascorbyl palmitate on linseed oil during in vitro gastrointestinal digestion was tackled. The advance of lipid oxidation was studied through the generation of oxidation compounds, the degradation of polyunsaturated fatty acyl chains and of gamma-tocopherol, by employing Proton Nuclear Magnetic Resonance. Supplements containing exclusively l-ascorbic acid enhanced the advance of linseed oil oxidation during digestion. This was evidenced by increased formation of linolenic-derived conjugated hydroxy-dienes and alkanals and by the generation of conjugated keto-dienes and reactive alpha,beta-unsaturated aldehydes, such as 4,5-epoxy-2-alkenals; moreover, gamma-tocopherol was completely degraded. Conversely, supplements composed of mixtures of ascorbic acid/salt with citric acid and carotenes, and of ascorbyl palmitate, protected linseed oil against oxidation and reduced gamma-tocopherol degradation. The study through Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry of the volatile compounds of the digests corroborated these findings. Furthermore, a decreased lipid bioaccessibility was noticed in the presence of the highest dose of l-ascorbic acid. Both the chemical form of Vitamin C and the presence of other ingredients in dietary supplements have shown to be of great relevance regarding oxidation and hydrolysis reactions occurring during lipid digestion.

Oxidative stability of extra-virgin olive oil enriched or not with lycopene. Importance of the initial quality of the oil for its performance during in vitro gastrointestinal digestion.

The performance of commercial non-enriched and lycopene-enriched extra-virgin olive oils (EVOO) during in vitro gastrointestinal digestion was studied in order to elucidate potential benefits of lycopene addition. Samples were analyzed before and after digestion by Proton Nuclear Magnetic Resonance (1H NMR) and Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry (SPME-GC/MS). EVOO samples differed in both main (oleic and linoleic acyl groups) and minor components (phenolic and oxidation compounds). Regardless of the presence of lycopene, all the samples reached a high degree of lipolysis and showed high stability towards oxidation under digestion conditions. Rather than oxidation reactions, the hydroperoxides initially present in the oil were reduced to more stable hydroxides. Likewise, hydroxy-diene isomerization from cis,trans to trans,trans occurred. Hence, the presumed antioxidant effect of lycopene was not noticed during in vitro digestion of EVOO. Similar experiments carried out with a more polyunsaturated oil (sunflower oil) indicated that lycopene slowed down the advance of oxidation slightly. However, in the case of EVOO, its initial quality prevailed over the slight antioxidant effect exerted by lycopene at the concentration present in commercial samples, determining the oxidation compound profile of the digests.

A study of the toxic effect of oxidized sunflower oil containing 4-hydroperoxy-2-nonenal and 4-hydroxy-2-nonenal on cortical TrkA receptor expression in rats.

Sunflower oil at a specific oxidation stage (when several oxygenated alpha,beta-unsaturated aldehydes are generated, mainly 4-hydroperoxy-trans-2-alkenals and 4-hydroxy-trans-2-alkenals), caused at 70 degrees C with aeration for 7 days, was administered intraperitoneally to rats. This oil was studied by means of solid phase micro-extraction followed by gas chromatography/mass spectrometry (SPME-GC/MS) and by proton nuclear magnetic resonance ((1)H-NMR). Oxidized sunflower oil (3 ml/kg/day) was administered to male Sprague-Dawley rats for 21 days. The control group was administered non-oxidized sunflower oil in the same volume and for the same duration as the experimental group. A significant decrease in the number of neural cells positively immunostained for TrkA receptor was detected in the frontal cortex of the experimental group, with respect to controls, suggesting both neuronal damage as well as a deficit in neuronal survival signalling at this level. This could lead to apoptosis of cholinergic neurons, which play a key role in memory and attention function. These results indicate that toxic substances present in the oxidized sunflower oil, among them 4-hydroxy-trans-2-nonenal (HNE) and 4-hydroperoxy-trans-2-nonenal (HPNE), could disrupt survival signalling of frontal cortex cholinergic neurons, which could lead to apoptosis and neurodegenerative diseases. In the case of humans, this fact reinforces the necessity of avoiding the re-utilization of oxidized sunflower oil, in order to contribute to long-term neurodegenerative diseases prevention.

Effect of adding alpha-tocopherol on the oxidation advance during in vitro gastrointestinal digestion of sunflower and flaxseed oils.

Few in vitro studies have tackled the effect of alpha-tocopherol on lipid oxidation during digestion, and discrepant results have been reported. As a result, the aim of this study was to elucidate whether the addition of alpha-tocopherol enhances or slows down the advance of oxidation that occurs during in vitro gastrointestinal digestion of polyunsaturated lipids. For this purpose, commercial sunflower and flaxseed oils (as models of omega-6 and omega-3 rich lipid systems, respectively) were in vitro digested in the absence or in the presence of this tocol at different concentrations (0.02%, 0.2% and 2%). Proton Nuclear Magnetic Resonance (1H NMR) and Solid Phase Microextraction followed by Gas Chromatography/Mass Spectrometry (SPME-GC/MS) were used to investigate in detail potential differences among the digests regarding lipolysis and oxidation level. Alpha-tocopherol addition did not affect the advance of lipolysis, whereas lipid oxidation was enhanced in a dose-dependent manner. In this regard, the increased degradation of polyunsaturated lipids and greater generation of primary and secondary oxidation products observed at higher concentrations of alpha-tocopherol confirmed this observation. Among the formed oxidation products, hydroperoxy-, hydroxy- and keto-dienes, as well as oxygenated alpha,beta-unsaturated aldehydes are worth mentioning. The in vitro bioaccessibility of added tocopherol was estimated to be very low, suggesting a notable transformation under the assayed conditions. Further in vivo studies are necessary to confirm this prooxidant activity of alpha-tocopherol during gastrointestinal digestion.

Evidence of the formation of light polycyclic aromatic hydrocarbons during the oxidation of edible oils in closed containers at room temperature.

Solid phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS) of the headspace composition of two sunflower oil samples was carried out; both samples were taken from the same original oil, stored for a prolonged time (112 months) in closed containers at room temperature under different air/oil volume ratios. Great differences in the headspace compositions of both samples were found due to the different oxidation levels reached. One of the most significant findings is that both contain monocyclic and light polycyclic aromatic hydrocarbons, the proportions of which are in line with the oxidation level of the sample. The determination of polycyclic aromatic compounds in the oil liquid matrix of both oil samples, carried out by means of a classical scheme of isolation, cleanup, separation, and quantification, showed that the concentrations of these compounds in the oil liquid phase also follow the oxidation degree reached by each sample, proving that this oxidation process at room temperature leads to the formation of these compounds.

Effects of different cooking methods on the lipids and volatile components of farmed and wild European sea bass (Dicentrarchus labrax).

European sea bass is very popular in the Mediterranean area, although very little is known about the possible different behaviours of farmed and wild samples during cooking. This study addresses the effect of microwave cooking, salt-crusted and conventional oven baking on the lipids and volatile profile of farmed and wild sea bass. Proton Nuclear Magnetic Resonance did not detect that hydrolysis or oxidation of lipidic components had taken place. However, Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry evidenced that polyunsaturated acyl group oxidation and Maillard-type reactions occurred to a very slight extent, yielding a wide variety of volatile odour-active compounds. Conventional baking enriched fish volatile profile to a higher extent than the other two techniques assayed. In fact, 15 Maillard reaction-derived compounds (pyrroles, alkylpyrazines, alkylthiophenes and 2-ethylpyridine) were only detected in oven-baked samples. Regardless of the cooking method applied, farmed sea bass showed a much richer aromatic profile than did wild samples, having 6-fold higher lipid content than the latter.

Detection of primary and secondary oxidation products by Fourier transform infrared spectroscopy (FTIR) and 1H nuclear magnetic resonance (NMR) in sunflower oil during storage.

The oxidation of sunflower oil, stored in closed receptacles at room temperature for a period of 10 years, was monitored using Fourier transform infrared spectroscopy (FTIR) and 1H nuclear magnetic resonance (NMR). The objective was to understand the evolution of the oxidation process in sunflower oil under the conditions above mentioned. These techniques provide information about the oxidative status of several oil samples and the primary and some of the secondary oxidation products formed in the oxidation process. The results obtained show that, under these conditions, sunflower oxidation takes place in a different way to that at higher temperatures with aeration. The 1H NMR spectra show that in the first oxidation stages of the process only hydroperoxides supporting cis, trans-conjugated double bonds are formed and that at more advanced stages hydroperoxides having trans, trans-conjugated double bonds are generated, with the latter always being in a smaller proportion than the former. In addition, the presence of hydroxy derivatives supporting cis, trans-conjugated double bonds among the primary oxidation compounds is shown for the first time. Also, from early oxidation stages onward and unlike the process at 70 degrees C with aeration, it is noticeable that 4-hydroxy- trans-2-alkenals are formed in much higher proportions than 4-hydroperoxy- trans-2-alkenals. This fact could be associated with the presence of hydroxy derivatives with cis, trans-conjugated double bonds among the primary oxidation products and the limited concentration of oxygen during the oxidation. Furthermore, relationships between some oxidation conditions and the oxidation level of the samples were statistically analyzed.

Effect of the presence of protein on lipolysis and lipid oxidation occurring during in vitro digestion of highly unsaturated oils.

The effect of the presence of ovalbumin and soy protein isolate on lipolysis and oxidation taking place during in vitro gastrointestinal digestion of slightly oxidized sunflower and flaxseed oils was addressed. The extent of lipolysis, the molar proportions of acyl groups/fatty acids after digestion, and the oxidation products formed were studied by Proton Nuclear Magnetic Resonance. The presence of proteins provoked a higher hydrolysis in triglycerides, a lower decrease of polyunsaturated chains, and a lower generation of oxidation compounds (conjugated dienes in chains having also hydroperoxy/hydroxy groups, epoxides and aldehydes); the formation of hydroxides was clearly favoured over that of hydroperoxides. Study of headspace composition by Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry confirmed that oxidation advanced to a lesser extent in the presence of protein. Thus, amino acids/peptides released during digestion may show antioxidant properties, affecting not only the extent of lipid oxidation, but also reactions pathways.

Behaviour of non-oxidized and oxidized flaxseed oils, as models of omega-3 rich lipids, during in vitro digestion. Occurrence of epoxidation reactions.

Fresh and partially oxidized flaxseed oil, as models of omega-3 rich lipids, were submitted to in vitro gastrointestinal digestion. Hydrolysis level, lipid composition and oxidative status of the samples before and after digestion were studied by Proton Nuclear Magnetic Resonance (1H NMR) and Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry (SPME-GC/MS). Although a great degree of lipolysis was reached in both kinds of samples after digestion, it was somewhat lower in the digests of oxidized flaxseed oil. The occurrence of lipid oxidation during digestion was evidenced by decreased unsaturated lipids and increased primary and secondary oxidation products, especially in oxidized samples. In these latter, linolenic-derived monoepoxy-octadecadienoates were the main oxidation products generated. SPME-GC/MS study showed the highest abundances of highly reactive alkadienals (C5-C10), alkatrienals (C9-C10) and linolenic-derived 4,5-epoxy-2-heptenals in the headspace of oxidized flaxseed oil digests. Volatile markers of Maillard-type reactions were also detected.

Effect of liquid smoking on lipid hydrolysis and oxidation reactions during in vitro gastrointestinal digestion of European sea bass.

The effect of smoking using liquid smoke flavourings on the hydrolysis and oxidation of European sea bass lipids during in vitro digestion was investigated. The techniques used were 1H NMR and SPME-GC/MS. The former proved that liquid smoking does not influence the extent of lipolysis, but prevents those lipid oxidation reactions that occur during in vitro digestion of unsmoked samples, giving rise to cis,trans-conjugated dienes associated with hydroperoxy/hydroxy groups. SPME-GC/MS corroborated the results obtained by 1H NMR in relation to the antioxidant effect of smoking under gastrointestinal conditions. Smoked sea bass digests showed lower abundances of volatile oxidation markers derived from omega-3 and omega-6 lipids than unsmoked ones. Moreover, the lowest values were found in the digests of sea bass samples smoked with the flavouring showing the highest phenolic content. For the first time, the bioaccessibility of smoke flavouring components was evidenced, some of them well-known for displaying antioxidant activity.

Polyunsaturated lipids and vitamin A oxidation during cod liver oil in vitro gastrointestinal digestion. Antioxidant effect of added BHT.

The extent of cod liver oil hydrolysis and oxidation during in vitro gastrointestinal digestion was investigated by Proton Nuclear Magnetic Resonance (1H NMR) and Solid Phase Microextraction-Gas Chromatography/Mass Spectrometry (SPME-GC/MS). These techniques evidenced the degradation of polyunsaturated ω-3 and ω-6 lipids and, for the first time, that of vitamin A, naturally present in cod liver oil. Cis,trans-conjugated dienes associated with hydroperoxides, as well as monoepoxides, cis,trans-2,4-alkadienals, 4-hydroperoxy- and 4-hydroxy-2-alkenals, and several vitamin A derived metabolites were generated. Moreover, the effect of the addition of the synthetic antioxidant 2,6-di-tert-butyl-hydroxytoluene (BHT) at 20 and 800ppm was tackled. Both techniques showed BHT to be efficient in limiting oxidation reactions during digestion, almost inhibiting them at 800ppm. Therefore, the simultaneous intake of antioxidants with cod liver oil should be considered, in order to increase polyunsaturated lipid and vitamin A bioaccessibility and avoid formation of toxic oxidation compounds like oxygenated alpha,beta-unsaturated aldehydes.

Direct study of minor extra-virgin olive oil components without any sample modification. 1H NMR multisupression experiment: A powerful tool.

Proton Nuclear Magnetic Resonance (1H NMR) was employed to study monovarietal commercial Spanish extra-virgin olive oils (EVOO) (Arbequina, Arroniz, Cornicabra, Hojiblanca and Picual). Each sample was analyzed by a standard pulse and by an experiment suppressing the main lipid signals, enabling the detection of signals of minor components. The aim was to determine the possibilities of both 1H NMR approaches to characterize EVOO composition, focusing on acyl groups, squalene, sterols, triterpene acids/esters, fatty alcohols, wax esters and phenols (lignans, tyrosol, hydroxytyrosol, oleocanthal, oleacein, oleokoronal, oleomissional, ligstrodials and oleuropeindials), and to determine hydrolysis and oxidation levels. The signal assignments (in deuterated chloroform) are thoroughly described, identifying for the first time those of the protons of esters of phytol and of geranylgeraniol. Correct signal assignment is fundamental for obtaining sound results when interpreting statistical data from metabolomic studies of EVOO composition and adulteration, making it possible to differentiate and classify oils.

Changes provoked by boiling, steaming and sous-vide cooking in the lipid and volatile profile of European sea bass.

This study aims to shed light on the changes provoked by boiling, steaming and sous-vide cooking on the lipids and volatile profile of farmed and wild European sea bass meat. None of the cooking techniques provoked changes due to hydrolytic or oxidation processes detectable by 1H NMR on sea bass lipids. The lipid profile of main and minor lipidic components was maintained after cooking. However, study by SPME-GC/MS evidenced that steaming and sous-vide cooking modified the volatile profile of sea bass meat, especially in farmed specimens. The compounds generated came from the occurrence, to a very small extent, of lipid and protein degradation. By contrast, boiling scarcely modified the initial characteristics of raw sea bass. Thus, from a sensory point of view and considering the odour-active compounds generated, steaming and sous-vide cooking provoked more noticeable changes than boiling, especially in farmed sea bass meat.

Fish in Vitro Digestion: Influence of Fish Salting on the Extent of Lipolysis, Oxidation, and Other Reactions.

A study of the various chemical reactions which take place during fish in vitro digestion and the potential effect of fish salting on their extent is addressed for the first time. Farmed European sea bass fillets, raw, brine-salted or dry-salted, were digested using a gastrointestinal in vitro model. Fish lipid extracts before and after digestion were analyzed by 1H NMR, and the headspace composition of the digestates was investigated by SPME-GC/MS. During digestion, not only lipolysis, but also fish lipid oxidation took place. This latter was evidenced by the generation of conjugated dienes supported on chains having also hydroperoxy- and hydroxy-groups (primary oxidation compounds), by the increase of volatile secondary oxidation products, and by the decrease of the antioxidant 2,6-di-tert-butyl-hydroxytoluene (BHT). Likewise, esterification and Maillard-type reactions also occurred. Salting, and especially dry-salting, enhanced all these reactions, except for lipolysis, during digestion.