Lipid oxidation in emulsions: New insights from the past two decades.

Lipid oxidation constitutes the main source of degradation of lipid-rich foods, including food emulsions. The complexity of the reactions at play combined with the increased demand from consumers for less processed and more natural foods result in additional challenges in controlling this phenomenon. This review provides an overview of the insights acquired over the past two decades on the understanding of lipid oxidation in oil-in-water (O/W) emulsions. After introducing the general structure of O/W emulsions and the classical mechanisms of lipid oxidation, the contribution of less studied oxidation products and the spatiotemporal resolution of these reactions will be discussed. We then highlight the impact of emulsion formulation on the mechanisms, taking into consideration the new trends in terms of emulsifiers as well as their own sensitivity to oxidation. Finally, novel antioxidant strategies that have emerged to meet the recent consumer's demand will be detailed. In an era defined by the pursuit of healthier, more natural, and sustainable food choices, a comprehensive understanding of lipid oxidation in emulsions is not only an academic quest, but also a crucial step towards meeting the evolving expectations of consumers and ensuring the quality and stability of lipid-rich food products.

Oleofoams: The impact of formulating air-in-oil systems from a lipid oxidation perspective.

Air-in-oil foams, or oleofoams, have a great potential for food applications as they can at least partially replace animal or hydrogenated fats, without compromising on textural properties. Yet, there are some challenges to tackle before they can largely be implemented for real-life applications. One of those is the lack of data regarding their oxidative stability. This is an important point to consider, as although using oils rich in polyunsaturated fatty acids (PUFAs) is highly desirable from a nutritional perspective, these fatty acids are particularly prone to oxidation, which leads to major degradations of food quality. This work thus aimed to investigate the oxidative stability of oleofoams prepared with omega-3 PUFA-rich vegetable oils (rapeseed or flaxseed oil) and various types of high melting point lipid-based oleogelators (stearic acid, glyceryl monostearate and stearyl alcohol) when incubated at room temperature. The physical structure and stability of the oleofoams was monitored by various techniques (visual observations, microscopy, DSC, NMR, SAXS and WAXS). Lipid oxidation was assessed by combined measurements of primary (conjugated diene hydroperoxides) and secondary (thiobarbituric acid reactive substances - TBARS) products. We found that the oxidative stability of oleofoams was higher compared to that of the corresponding bulk oil. This protective effect was also found when the oil was simply mixed with the oleogelator without incorporation of air bubbles (i.e., forming an oleogel), and was somewhat modulated depending on the type of oleogelator. These results suggest that oleogelators and the structural changes that they induce limit the cascaded propagation of lipid oxidation in oil-continuous matrices, which is promising in the perspective of future applications.

Effect of some local plant extracts on fatty acid composition of fish (Alestes baremoze) during smoking and sun drying in the Far-North region of Cameroon.

The objective of this study was to assess the antioxidant activities of three plant extracts (Moringa oleifera leaves, Xylopia aethiopica fruits, and Allium cepa leaves) and to evaluate their effects on the preservation of fish polyunsaturated fatty acids (PUFAs) during smoking and sun-drying processes. PUFAs are highly prone to oxidation during fish processing. The plant extracts were analyzed for their polyphenol contents and were evaluated for their total antiradical capacity. The polyphenol components of each plant were characterized. The hydroethanolic and aqueous extracts were added to the fish at concentrations of 3, 6, 9, and 12 g/L and 10, 20, 30, and 40 g/L, respectively. Butylated hydroxytoluene (BHT) was used as a positive control at a concentration of 2 g/L to compare the antioxidant effects of the plant extracts. The treated fish was subjected to smoking or sun drying and the fatty acid composition of the fish lipid extract was assessed. The results showed that the total polyphenolic, flavonoid, and tannin contents varied significantly from one plant extract to the other (p < .05). The radical scavenging and FRAP increased significantly with the concentration of the plant extracts (p < .05). An HPLC analysis of the extracts led to the preliminary identification of four hydroxycinnamic acids in M. oleifera and X. aethiopica, one anthocyanin and one flavone glycoside in M. oleifera, and four flavan-3-ols in X. aethiopica. Moreover, eight flavonols were preliminarily identified in the three plants. Compared to the control product, these plant extracts significantly protected fish PUFAs from oxidation (p < .05). The aqueous extract of A. cepa at 40 g/L better preserved omega-3 in fish during smoking and sun drying than the control product. Incorporating the three plant extracts during smoking and sun-drying processes can effectively preserve the PUFAs in fish. Therefore, these plants are viable sources of natural antioxidants in the preservation of fish products.

In vitro digestion of high-lipid emulsions: towards a critical interpretation of lipolysis.

Investigating the gastrointestinal fate of food emulsions is critical to unveil their nutritional relevance. To this end, the protocol standardized by COST INFOGEST 2.0 is meaningful for guiding in vitro digestion experiments. In contrast with studies addressing emulsions with low dispersed phase volume fraction (φ 0.05-0.1), we presently raise some points for a proper interpretation of the digestibility of emulsions with high lipid content using the pH-stat method. Oil-in-water high internal phase emulsions (HIPEs) were submitted to gastric pre-lipolysis with the addition of rabbit gastric lipase (RGE). Commercial mayonnaise (φ 0.76) was systematically diluted (φ 0.025, 0.05, 0.1, 0.15, 0.25, 0.4, and 0.76) to cover a wide range of enzyme-to-lipid ratios (8.5-0.3 U per µmol for RGE and 565.1-18.6 U per µmol for pancreatin, in the gastric and intestinal phases, respectively). Lipolysis was tracked either by fatty acid titration (NaOH titration) or completed by analysis of lipid classes and fatty acid composition. Gastric lipase resulted in substantial lipid hydrolysis, reaching 20 wt% at low lipid fractions (φ 0.025 and 0.05). Likewise, the kinetics and extent of lipolysis during intestinal digestion were modulated by the enzyme-to-substrate ratio. A logarithmic relationship between lipid hydrolysis and lipid concentration was observed, with a very limited extent at the highest lipid content (φ 0.76). A holistic interpretation relying on FFA titration and further evaluation of all lipolytic products appears of great relevance to capture the complexity of the effects involved. Overall, this work contributes to rationally and critically evaluating the outcomes of static in vitro experiments of lipid digestion.

Dietary oxidized n-3 PUFA induce oxidative stress and inflammation: role of intestinal absorption of 4-HHE and reactivity in intestinal cells.

Dietary intake of long-chain n-3 PUFA is now widely advised for public health and in medical practice. However, PUFA are highly prone to oxidation, producing potentially deleterious 4-hydroxy-2-alkenals. Even so, the impact of consuming oxidized n-3 PUFA on metabolic oxidative stress and inflammation is poorly described. We therefore studied such effects and hypothesized the involvement of the intestinal absorption of 4-hydroxy-2-hexenal (4-HHE), an oxidized n-3 PUFA end-product. In vivo, four groups of mice were fed for 8 weeks high-fat diets containing moderately oxidized or unoxidized n-3 PUFA. Other mice were orally administered 4-HHE and euthanized postprandially versus baseline mice. In vitro, human intestinal Caco-2/TC7 cells were incubated with 4-hydroxy-2-alkenals. Oxidized diets increased 4-HHE plasma levels in mice (up to 5-fold, P < 0.01) compared with unoxidized diets. Oxidized diets enhanced plasma inflammatory markers and activation of nuclear factor kappaB (NF-κB) in the small intestine along with decreasing Paneth cell number (up to -19% in the duodenum). Both in vivo and in vitro, intestinal absorption of 4-HHE was associated with formation of 4-HHE-protein adducts and increased expression of glutathione peroxidase 2 (GPx2) and glucose-regulated protein 78 (GRP78). Consumption of oxidized n-3 PUFA results in 4-HHE accumulation in blood after its intestinal absorption and triggers oxidative stress and inflammation in the upper intestine.

n-3 PUFA prevent metabolic disturbances associated with obesity and improve endothelial function in golden Syrian hamsters fed with a high-fat diet.

Glucose intolerance and dyslipidaemia are independent risk factors for endothelium dysfunction and CVD. The aim of the present study was to analyse the preventive effect of n-3 PUFA (EPA and DHA) on lipid and carbohydrate disturbances and endothelial dysfunction. Three groups of adult hamsters were studied for 20 weeks: (1) control diet (Control); (2) high-fat diet (HF); (3) high-fat diet enriched with n-3 PUFA (HFn-3) groups. The increase in body weight and fat mass in the HF compared to the Control group (P < 0.05) was not found in the HFn-3 group. Muscle TAG content was similar in the Control and HF groups, but significantly lower in the HFn-3 group (P = 0.008). Glucose tolerance was impaired in the HF compared to the Control group, but this impairment was prevented by n-3 PUFA in the HFn-3 group (P < 0.001). Plasma TAG and cholesterol were higher in the HF group compared to the Control group (P < 0.001), but lower in the HFn-3 group compared to the HF group (P < 0.001). HDL-cholesterol was lower in the HFn-3 group compared to the Control and HF groups (P < 0.0005). Hepatic secretion of TAG was lower in the HFn-3 group compared to the HF group (P < 0.005), but did not differ from the Control group. Hepatic gene expression of sterol regulatory element-binding protein-1c, diacylglycerol O-acyltransferase 2 and stearyl CoA desaturase 1 was lower in the HFn-3 group, whereas carnitine palmitoyl transferase 1 and scavenger receptor class B type 1 expression was higher (P < 0.05). In adipocytes and adipose macrophages, PPARγ and TNFα expression was higher in the HF and HFn-3 groups compared to the Control group. Endothelium relaxation was higher in the HFn-3 (P < 0.001) than in the HF and Control groups, and was correlated with glucose intolerance (P = 0.03) and cholesterol (P = 0.0003). In conclusion, n-3 PUFA prevent some metabolic disturbances induced by high-fat diet and improve endothelial function in hamsters.

Rapid assessment of fatty acyls chains of phospholipids and plasmalogens by atmospheric pressure chemical ionization in positive mode and high-resolution mass spectrometry using in-source generated monoacylglycerol like fragments intensities.

We recently published a new concept using monoacylglycerol-like fragments [MG+H-H2O]+ (ions B) produced in-source by atmospheric pressure photoionization in positive mode and high-resolution mass spectrometry for the determination of the fatty acyl (FA) composition of triacylglycerols (TGs) from plant oils. This study extends the concept to the phospholipids (PLs) category and shows that the APCI+ source can also be used. Moreover, the coupling with NP-LC allows to simultaneously analyze different PLs classes in the same sample. We compared the relative intensities of the ions B produced in-source to the % composition of FAs determined by GC-FID. In the case of PLs from natural extracts composed exclusively of diacyl-PLs, the relative intensities of ions B are close to the % of the FAs obtained by GC-FID. This approach is not directly useable for extracts containing plasmalogens (P-PLs). For these PLs, acidic hydrolysis by HCl fumes allows hydrolyzing selectively vinyl ether functions to form lyso-PLs. The analysis of hydrolyzed extracts makes it possible to obtain the composition of P-PLs FAs thanks to the lyso-PLs thus formed, while the diacyl-PLs composition remains unchanged. Unlike GC-FID FAs determination, this approach allows a distinction between the diacyl-PLs and P-PLs FAs composition. We also found that the ion B intensities were consistent among the PL classes (PG, PE, PA, PI, CL, PS and PC) and lyso- forms (LPE and LPC). In the case of the diacyl-PLs extracts analyzed, no statistically significant differences were found between the PLs FAs compositions calculated from ion B intensities and the corresponding GC-FID data. A weighting coefficient was applied to correct ion B intensities issued from polyunsaturated FAs with three or more double bonds. The fatty alkenyls composition of P-PLs could also be calculated from the % intensities of specific ions.

Monitoring oxidation during the storage of pressure-treated cooked ham and impact on technological attributes.

High-pressure processing is a post-processing preservation method commonly used on meat products. However, it can affect the structural properties and the physico-chemical properties of the meat. The aim of this study was to compare the physical properties, lipid and protein oxidation of control and treated (500 MPa, 20 °C, 5 min) cooked ham during subsequent storage (21 days at 4 °C). High pressure processing induced increase of hardness and syneresis after 7 days of storage. The redness (a*) was slightly affected by the high pressure treatment but not the lightness (L*) and the yellowness (b*). However, the fluctuation of color was not clearly visible. Evaluation of primary (conjugated dienes) and secondary (malondialdehyde MDA and thiobarbituric reactive substances TBA-RS) lipid oxidation products showed that pressure increases oxidation of lipids. Whereas, high pressure processing had no immediate effect on MDA and TBA-RS content, higher amount compared to control were observed during the refrigerated storage. This lipid oxidation could be due to the release of prooxidant iron from hemoproteins after the high pressure treatment. Finally, the determination of free and accessible thiols showed that the high pressure treatment leads to a protein oxidation.

Modeling the fragmentation patterns of triacylglycerides in mass spectrometry allows the quantification of the regioisomers with a minimal number of standards.

Mass spectrometry allows the relative quantification of the regioisomers of triacylglycerides by the calibration of their fragmentation patterns. However, due to the plethora of regioisomers of triacylglycerides, calibration with every standard is not feasible. An analytical challenge in the field is the prediction of the fragmentation patterns of triacylglycerides to quantify their regioisomers. Thus, we aimed to model these fragmentation patterns to quantify the regioisomeric composition, even for those without commercially available standards. In a first step, we modeled the fragmentation patterns of the regioisomers of triacylglycerides obtained from different published datasets. We found the same qualitative trends of fragmentation beyond differences in the type of adduct in these datasets (both [M+NH4]+ and [M+H]+), and the type of instrument (orbitrap, Q-ToF, ion-trap, single quadrupole, and triple quadrupole). However, the quantitative trends of fragmentation were adduct and instrument specific. From these observations, we modeled quantitatively the common trends of fragmentation of triacylglycerides in every dataset. In a second step, we applied this methodology on a Synapt G2S Q-ToF to quantify the regioisomers of triacylglycerides in sunflower and olive oils. The results of our quantification were in good agreement with previous published quantifications of triacylglycerides, even for regioisomers that were not present in the training dataset. The species with more than two highly unsaturated fatty acids (arachidonic, eicosapentaenoic, and docosahexaenoic acids) showed a complex behavior and lower predictability of their fragmentation patterns. However, this framework presents the capacity to model this behavior when more data are available. It would be also applicable to standardize the quantification of the regioisomers of triacylglycerides in an inter-laboratory ring study.

Modifications of protein-related compounds of beef minced meat treated by high pressure.

In this study, we evaluated the modifications of the protein-related compounds of minced beef treated with high pressure, including refrigeration after treatment. The free amino acid content, protein carbonyls and free thiol groups were assessed. High pressure up to 200 MPa had a significant effect. Above 300 MPa, irreversible structural changes occurred, with an increase in the protein oxidation products and a modification of the amounts of amino acids after 14 days of storage. Protein carbonyls and the free thiols were correlated with the free amino acids. These results showed that protein modifications under pressure result from both conformational and chemical changes, possibly associated with lipid changes under high-pressure treatment.

The initial characteristics of marine oil emulsions and the composition of the media inflect lipid oxidation during in vitro gastrointestinal digestion.

This work aimed at evaluating the effect of simulated digestive fluids, interface and lipid droplet sizes on the oxidation of oil-in-water emulsions containing long chain n-3 fatty acyls. Emulsions stabilised by a protein or by phosphatidyl-choline/Tween 80 were submitted to gastro-intestinal in vitro conditions in presence of metmyoglobin. The gastric phase was characterised by a decrease of tocopherol amounts and moderate O2 uptake and aldehyde formation. Oxidation developed further during the intestinal phase, with tocopherols tending to zero, oxygen uptake and production of aldehydes at potentially toxic concentrations. The simulated digestive fluids reduced oxygen uptake and MDA formation only during the intestinal step of the phospholipid-stabilised emulsion. Quantitative losses of PUFA (e.g. EPA, DHA) were less than 10% even significant at the end of the digestion.

Maternal supplementation with n-3 long chain polyunsaturated fatty acids during perinatal period alleviates the metabolic syndrome disturbances in adult hamster pups fed a high-fat diet after weaning.

Perinatal nutrition is thought to affect the long-term risk of the adult to develop metabolic syndrome. We hypothesized that maternal supplementation with eicosapentaenoic acid and docosahexaenoic acid during pregnancy and lactation would protect offspring fed a high-fat diet from developing metabolic disturbances. Thus, two groups of female hamsters were fed a low-fat control diet, either alone (LC) or enriched with n-3 long chain polyunsaturated fatty acids (LC-PUFA) (LO), through the gestational and lactation periods. After weaning, male pups were randomized to separate groups that received either a control low-fat diet (LC) or a high-fat diet (HC) for 16 weeks. Four groups of pups were defined (LC-LC, LC-HC, LO-LC and LO-HC), based on the combinations of maternal and weaned diets. Maternal n-3 LC-PUFA supplementation was associated with reduced levels of basal plasma glucose, hepatic triglycerides secretion and postprandial lipemia in the LO-HC group compared to the LC-HC group. Respiratory parameters were not affected by maternal supplementation. In contrast, n-3 LC-PUFA supplementation significantly enhanced the activities of citrate synthase, isocitrate dehydrogenase and α-ketoglutarate dehydrogenase compared to the offspring of unsupplemented mothers. Sterol regulatory element binding protein-1c, diacylglycerol O-acyltransferase 2, fatty acid synthase, stearoyl CoA desaturase 1 and tumor necrosis factor α expression levels were not affected by n-3 LC-PUFA supplementation. These results provide evidence for a beneficial effect of n-3 LC-PUFA maternal supplementation in hamsters on the subsequent risk of metabolic syndrome. Underlying mechanisms may include improved lipid metabolism and activation of the mitochondrial oxidative pathway.

In vitro digestion of fish oils rich in n-3 polyunsaturated fatty acids studied in emulsion and at the oil-water interface.

The in vitro digestion of ß-lactoglobulin stabilized emulsions rich in the n-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), was studied using several physicochemical techniques. Artificial media for the mouth, stomach and small intestine were used in a sequential static in vitro digestion method. Different sizing techniques were compared to follow the droplet size during the digestion steps, including diffusing wave spectroscopy (DWS) which allowed direct measurements on undiluted emulsions. Titration of fatty acids confirmed that the digestion of such emulsified fish oils is partial. The study of the digestion at the oil-water interface using tensiometry revealed specific affinities between lipids and proteins. These could explain the emulsion and the single droplet lipolysis. Nevertheless, by comparing our results to a previous study on fish oil lipolysis, we identified two other important factors. Those were the aqueous solubility and the rate of hydrolysis of the individual fatty acids, the emulsion with the most soluble and hydrolysable ones being digested more quickly.

Odour quality of spray-dried hens' egg powders: the influence of composition, processing and storage conditions.

This study aimed to determine whether compositional or processing parameters have an influence on the odour quality of egg powders. The parameters tested were: whole egg vs. egg yolk, polyunsaturated fatty acid (PUFA) enrichment, spray-drying temperature (160°C vs. 180°C), production scale (industrial vs. pilot plant), storage temperature (15°C vs. 30°C) and time (1, 2, 4 and 8 months). The quality of egg powders was evaluated by sensory analysis using free sorting, and by gas chromatography coupled to mass spectrometry and olfactometry. PUFA enrichment and spray-drying temperature do not affect the odour of egg yolk powders. There are significant differences between the odour of whole-egg and egg-yolk powders as well as between powders produced on an industrial scale or in a pilot plant. An increase in the odour intensity of egg powders was observed during storage, while unpleasant odours were perceived when the egg powders were stored at 30°C.

n-3 PUFA added to high-fat diets affect differently adiposity and inflammation when carried by phospholipids or triacylglycerols in mice.

BACKGROUND: Dietary intake of n-3 polyunsaturated fatty acids (PUFA) is primarily recognized to protect against cardiovascular diseases, cognitive dysfunctions and the onset of obesity and associated metabolic disorders. However, some of their properties such as bioavailability can depend on their chemical carriers. The objective of our study was to test the hypothesis that the nature of n-3 PUFA carrier results in different metabolic effects related to adiposity, oxidative stress and inflammation. METHODS: 4 groups of C57BL/6 mice were fed for 8 weeks low fat (LF) diet or high-fat (HF, 20%) diets. Two groups of high-fat diets were supplemented with long-chain n-3 PUFA either incorporated in the form of phospholipids (HF-ω3PL) or triacylglycerols (HF-ω3TG). RESULTS: Both HF-ω3PL and HF-ω3TG diets reduced the plasma concentrations of (i) inflammatory markers such as monocyte chemoattractant protein-1 (MCP-1) and interleukin 6 (IL-6), (ii) leptin and (iii) 4-hydroxy-2-nonenal (4-HNE), a marker of n-6 PUFA-derived oxidative stress compared with the control HF diet. Moreover, in both HF-ω3PL and HF-ω3TG groups, MCP-1 and IL-6 gene expressions were decreased in epididymal adipose tissue and the mRNA level of gastrointestinal glutathione peroxidase GPx2, an antioxidant enzyme, was decreased in the jejunum compared with the control HF diet. The type of n-3 PUFA carrier affected other outcomes. The phospholipid form of n-3 PUFA increased the level of tocopherols in epididymal adipose tissue compared with HF-ω3TG and resulted in smaller adipocytes than the two others HF groups. Adipocytes in the HF-ω3PL and LF groups were similar in size distribution. CONCLUSION: Supplementation of mice diet with long-chain n-3 PUFA during long-term consumption of high-fat diets had the same lowering effects on inflammation regardless of triacyglycerol or phospholipid carrier, whereas the location of these fatty acids on a PL carrier had a major effect on decreasing the size of adipocytes that was not observed with the triacyglycerol carrier. Altogether, these results would support the development functional foods containing LC n-3 PUFA in the form of PL in order to prevent some deleterious outcomes associated with the development of obesity.

Gastric conditions control both the evolution of the organization of protein-stabilized emulsions and the kinetic of lipolysis during in vitro digestion.

During digestion, lipids undergo modifications of their colloidal and molecular structures, which depend on the digestive conditions and the composition of the digestive juices. The aim of this work was to evaluate whether gastric pH and pepsin modulate the colloidal evolution and the bioacessibility of fatty acids of an oil-in-water emulsion stabilized by a protein during in vitro digestion. The fate of BSA-stabilized rapeseed oil-in-water emulsion during gastric phase at pH 2.5 or 4.0 with or without pepsin and its consequences on intestinal lipolysis was measured in the simulated gastric and duodenal conditions. The pH had limited impact but pepsin favoured flocculation and coalescence of the droplets, modulating the early stage of lipolysis but not its final extent.