Encarnación Goicoechea

Toxic Oxygenated α,β-Unsaturated Aldehydes and their Study in Foods: A Review

The oxidation of lipids containing polyunsaturated omega-3 or omega-6 acyl groups, such as docosahexenoic, eicosapentenoic, linolenic, arachidonic, or linoleic groups, and of the corresponding fatty acids, generates among other compounds α,β -unsaturated aldehydes supporting different functional groups containing oxygen, which can be named oxygenated α,β -unsaturated aldehydes (Oα β UAs). These compounds can be produced in cells and tissues of living organisms or in foods during processing or storage, and from these latter can be absorbed through the diet. In the last few years, Oα β UAs are receiving a great deal of attention because they are being considered as possible causal agents of numerous diseases, such as chronic inflammation, neurodegenerative diseases, adult respiratory distress syndrome, atherogenesis, diabetes, and different types of cancer. This review deals with the nature of the different kinds of Oα β UAs detected until now, their reactivity and consequent biological activity; the several pathways proposed for their formation; the current knowledge about the influence of both oxidative conditions and lipids nature in the rate of formation and yield of each kind of Oα β UAs in edible oils; the methods described until now to determine the presence in foods of some of these compounds, such as 4-hydroxy-trans-2-nonenal, 4-hydroxy-trans-2-octenal, 4-hydroxy-trans-2-hexenal and 4-oxo-trans-2-hexenal; and finally, the levels found of some of them in several foods.

Analysis of Hydroperoxides, Aldehydes and Epoxides by 1H Nuclear Magnetic Resonance in Sunflower Oil Oxidized at 70 and 100 °C

A global study of sunflower oil oxidation at two different temperatures (100 and 70 degrees C) with aeration was carried out by (1)H nuclear magnetic resonance ((1)H NMR), paying attention not only to the degradation of the main components but also to the formation of others. The functional groups of compounds monitored simultaneously during the oxidation were, in addition to acyl groups, the following: hydroperoxides, conjugated dienic systems of hydroperoxy acyl groups, aldehydes including the genotoxic and cytotoxic oxygenated alpha,beta-unsaturated aldehydes, and mono- and diepoxides. Chemical shifts of protons of all mentioned groups were given. Differences between both oxidation processes were found from the initial stages to the end. The formation of diepoxides in the oxidation process of edible oils was shown for the first time, as was the influence of temperature on the formation of epoxy acyl groups, some of these latter related to leukotoxin and isoleukotoxin.

Quality of farmed and wild sea bass lipids studied by 1H NMR: Usefulness of this technique for differentiation on a qualitative and a quantitative basis

Proton Nuclear Magnetic Resonance ((1)H NMR) spectroscopy was employed to study the lipids of farmed and wild European sea bass (Dicentrarchus labrax). This technique provided a great deal of detailed information on sea bass lipids composition and, once the spectra signals have been assigned, the simple observation of the spectra enables one to distinguish cultured from wild fish. This distinction is possible due to the presence in the former of high proportions of diunsaturated acyl groups, mainly linoleic. Furthermore, new approaches for quantitative characterization of sea bass lipids were developed. For the same season wild sea bass lipids contain not only higher molar percentages of omega-3 and of docosahexaenoic (DHA), but also higher concentrations of phosphatidylcholine and of cholesterol than farmed ones. However, in general, the absolute content of DHA and of eicosapentaenoic plus arachidonic (EPA+ARA) acyl groups as well as of phosphatidylcholine and cholesterol, were reasonably higher in farmed than in wild sea bass fillets, due to the higher total lipid content of the former. The presence of omega-1 acyl groups in all farmed samples and in some of the wild specimens was shown. As far as we know, this is the first time that omega-1 acyl groups have been identified and quantified in fish lipids by (1)H NMR.

Characterization of Cod Liver Oil by Spectroscopic Techniques. New Approaches for the Determination of Compositional Parameters, Acyl Groups, and Cholesterol from 1H Nuclear Magnetic Resonance and Fourier Transform Infrared Spectral Data

Six samples of cod liver oil were studied using Fourier transform infrared (FTIR) spectroscopy and (1)H nuclear magnetic resonance ((1)H NMR). These techniques provide information simply and rapidly about the global features of the cod liver oil main components, showing their potential as routine techniques for evaluating certain parameters of the quality of the cod liver oil. FTIR spectroscopy provides information about the molar percentage of polyunsaturated acyl groups in the sample and also about the ratio between unsaturated and saturated structures. (1)H NMR provides information about the proportions or concentrations of certain acyl groups and also of some minor compounds such as cholesterol. Both techniques are simple and fast. New approaches are presented to evaluate the molar proportions or concentrations of some acyl groups such as the molar percentages of omega-3, docosahexaenoic, and eicosapentaenoic acyl groups; furthermore, some novel approaches for evaluating the molar percentages of unsaturated and saturated acyl groups are also given. Results obtained from both spectroscopic techniques are in total agreement.

Fate in digestion in vitro of several food components, including some toxic compounds coming from omega-3 and omega-6 lipids

In this study it was proved the formation of oxygenated alpha,beta-unsaturated aldehydes (OαβUAs) of 6, 7, 9 and 10 carbon atoms during the thermal treatment (190°C with aeration) of a commercial vegetable oil rich in omega-3 and omega-6 acyl groups, which also contained small amounts of added proteins and carbohydrates to produce barbecue aroma when heated. The OαβUAs detected by Solid Phase Microextraction (SPME) followed by Gas Chromatography/Mass Spectrometry (GC/MS) were: 4-hydroxy-2-hexenal, 4-oxo-2-hexenal and 4,5-epoxy-2-heptenals, coming from omega-3 acyl groups; and 4-hydroxy-2-nonenal, 4-oxo-2-nonenal and 4,5-epoxy-2-decenals, coming from omega-6 acyl groups. Mixtures of this oil, either thermodegraded or not, with standard food were submitted to an in vitro digestion model. The study of the digestion products obtained revealed that OαβUAs remained unaltered, being bioaccessible in the gastrointestinal tract and so able to reach the systemic circulation. Besides, it was evidenced that during digestion Maillard, esterification and oxidation reactions take place.

A Review of Thermo-Oxidative Degradation of Food Lipids Studied by 1H NMR Spectroscopy: Influence of Degradative Conditions and Food Lipid Nature

This review summarizes present-day knowledge provided by proton nuclear magnetic resonance (1H NMR) concerning food lipid thermo-oxidative degradation. The food lipids considered include edible oils and fats of animal and vegetable origin. The thermo-oxidation processes of food lipids of very different composition, occurring at low, intermediate, or high temperatures, with different food lipid surfaces exposed to oxygen, are reviewed. Mention is made of the influence of both food lipid nature and degradative conditions on the thermo-oxidation process. Interest is focused not only on the evolution of the compounds that degrade, but also on the intermediate or primary oxidation compounds formed, as well as on the secondary ones, from both qualitative and quantitative points of view. Very valuable qualitative and quantitative information is provided by 1H NMR, which can be useful for metabolomic and lipidomic studies. The chemical shift assignments of spectral signals of protons of primary (hydroperoxides and hydroxides associated with conjugated dienes) and secondary, or further (aldehydes, epoxides, among which 9,10-epoxy-12-octadecenoate [leukotoxin] can be cited, alcohols, ketones) oxidation compounds is summarized. It is worth noting the ability of 1H NMR to detect toxic oxygenated α,β-unsaturated aldehydes, like 4-hydroperoxy-, 4,5-epoxy-, and 4-hydroxy-2-alkenals, which can be generated in the degradation of food lipids having omega-3 and omega-6 polyunsaturated groups in both biological systems and foodstuffs. They are considered as genotoxic and cytotoxic, and are potential causative agents of cancer, atherosclerosis, and Parkinsons and Alzheimers diseases.

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.

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.

Use of an in Vitro Digestion Model To Study the Bioaccessibility of 4-Hydroxy-2-nonenal and Related Aldehydes Present in Oxidized Oils Rich in Omega-6 Acyl Groups

Mixtures of either sunflower oil or thermodegraded sunflower oil and a standard meal were submitted to an in vitro digestion model. The same experiment was carried out with fluid deep-frying fat and thermodegraded fluid deep-frying fat. The thermodegradation of the oil and fat was provoked by submitting them to 190 degrees C with aeration in a convection oven, and the presence in the headspace of the thermodegraded oil and fat of oxygenated alpha,beta-unsaturated aldehydes (OalphabetaUAs), such as 4-hydroxy-2-nonenal (HNE), 4-oxo-2-nonenal (ONE), and 4,5-epoxy-2-decenal (EDE), was monitored by solid phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS). The digestion products were separated by centrifugation in a lipidic phase, an aqueous phase, and a pellet phase. The headspace of these three phases was also studied by SPME/GC-MS to check if the toxic and very reactive OalphabetaUAs above-mentioned remained unaltered after the in vitro digestion process or if they had reacted with the various compounds present in the digestion products, so disappearing from the samples. With the same aim the extract in ethyl acetate of the aqueous and pellet phases, and of the lipidic phase after dilution, were studied by GC-MS. All results obtained showed that a certain proportion of the toxic OalphabetaUAs remains unaltered after digestion, dispersed in the three phases above-mentioned, and thus are bioaccessible in the gastrointestinal tract and so could reach the systemic circulation. Compounds that may originate in Maillard type reactions (2-pentylpyridine) are found among digestion products, proving that these reactions are possible in this process if adequate substrates are present. In addition, it has been shown that toxic metabolites from the synthetic antioxidant BHT, present in fat before digestion, remain unaltered after this process and could reach the systemic circulation.

Usefulness of 1H NMR in assessing the extent of lipid digestion

Proton Nuclear Magnetic Resonance ((1)H NMR) is proved to be, for the first time, a very useful technique in monitoring the extent of lipid hydrolysis in digestion processes. Sunflower oil and minced fish flesh, as model foods, were subjected to different in vitro digestion experiments and the lipolysis levels reached were evaluated using (1)H NMR spectral data. Simple observation of the spectra gives very valuable information about the extent of the lipolysis and enables a rapid discrimination among samples having different hydrolysis degree. Equations were developed to quantify all the lipolytic products, and either referred to acyl groups plus fatty acids, or to glyceryl structures. The main hydrolysis products were 1,2-diglycerides, 2-monoglycerides, glycerol and fatty acids, although small proportions of 1,3-diglycerides and of 1-monoglycerides were also found. With this methodology, determination of the extent of lipid digestion in its different definitions can be made. It has been shown that these definitions are not equivalent, which is evidence for the need for a consensus in this regard.