Oxidative Stability of Whey Protein-stabilized Oil-in-water Emulsions at pH 3: Potential ω-3 Fatty Acid Delivery Systems (Part B)

ABSTRACT: Consumption of omega-3 (ω-3) fatty acids is beneficial for human health. Incorporation of ω-3 fatty acids into functional foods is limited by their high susceptibility to oxidative degradation. Oil-in-water emulsions may be a more effective method to deliver ω-3 fatty acids into functional foods. Protein-stabilized oil-in-water emulsions at pH values below the isoelectric point of the protein produce cationic emulsion droplets that decrease the oxidation of lipids by decreasing iron-lipid interactions. This research showed that whey protein isolate (WPI)-stabilized algal oil emulsions at pH 3.0 had good physical and oxidative stability after pasteurization. Addition of ethylenediaminetetraacetic acid     

Influence of heat processing and calcium ions on the ability of EDTA to inhibit lipid oxidation in oil-in-water emulsions containing omega-3 fatty acids

The nutritional benefits of ω-3 fatty acids make them excellent candidates as functional food ingredients if problems with oxidative rancidity can be overcome. Oil-in-water emulsions were prepared with 2% salmon oil, stabilized by 0.2% Brij 35 at pH 7. To determine the effects of heating (50–90 °C), ethylenediaminetetraacetic acid (EDTA), and calcium on the oxidative and physical stability of salmon oil-in-water emulsions, particle size, thiobarbituric acid reactive substances (TBARS), and lipid hydroperoxides were measured. The heat-processed emulsions showed no significant difference, in particle size, TBARS or hydroperoxides during storage, from unheated emulsions. Above 2.5 μM, EDTA dramatically decreased lipid oxidation in all samples. Addition of calcium to emulsions containing 7.5 μM EDTA significantly increased both TBARS and hydroperoxide formation when calcium concentrations were 2-fold greater than EDTA concentrations. These results indicate that heat-processed salmon oil-in-water emulsions with high physical and oxidative stability could be produced in the presence of EDTA.     

Effect of antioxidants on stabilization of meat products fortified with n-3 fatty acids

The effects of an n-3 oil emulsion, with and without added antioxidants, on lipid oxidation in n-3 polyunsaturated fatty acid (PUFA)-fortified meat products were studied. An emulsion of n-3 PUFAs was prepared (25% algal oil, 2.5% whey protein isolates, 10mM sodium citrate, 0.2% potassium sorbate, 500ppm of 70% mixed tocopherols, 100μM EDTA, pH 3, pasteurized at 75°C for 30min) and incorporated into fresh ground turkey, and fresh pork sausage (20% fat) to achieve a concentration of 500mg n-3 PUFA/110g meat. An antioxidant combination containing rosemary (0.2% w/w; radical quencher), citrate (0.5% w/w; sequestrant) and erythorbate (1g/kg product; reductant) was prepared and incorporated into ground turkey patties (5cm dia, 1.5cm thick) or fresh pork sausages (5cm dia, 1.5cm thick). Meat products were stored at 4°C or -18°C and analyzed for color (L*, a*, b* values), lipid oxidation (TBARS and lipid hydroperoxides) and n-3 PUFA profile. a* Values of refrigerated ground turkey patties decreased with storage, and an antioxidant combination effect was observed after 4 days (P<0.05). For fresh pork sausages at 4°C, control+antioxidant (CON+ANTI), and n-3+antioxidant (n-3+ANTI) groups showed greater a* values than controls (CON) indicating that the antioxidant combination stabilized meat color. TBARS and lipid hydroperoxides of both n-3 PUFA-enhanced meat products increased with storage (P<0.05); there were no significant changes in TBARS or lipid hydroperoxides for treatments containing the antioxidant combination (P<0.05). The actual level of n-3 PUFA incorporation in both meat products was greater than 87%; n-3 PUFA concentrations did not change within any treatment during storage (P>0.05). These results provide support for including antioxidant protection in n-3 PUFA fortified meat products.     

Oxidative stability of frozen-stored raw pork chops, chill-stored pre-frozen raw pork chops, and frozen-stored pre-cooked sausages in relation to dietary CuSO4, rapeseed oil and vitamin E

 Danish Landrace× Danish Yorkshire female pigs were fed either a standard diet or a standard diet enriched with 6% rapeseed oil and supplemented with increasing amounts of vitamin E (0, 100 or 200 mg all-rac-α-tocopheryl acetate/kg feed) and copper (0, 35 or 175 mg CuSO₄/kg feed), and the effect of dietary regimen on the oxidative stability of (1) frozen-stored raw pork chops packed in atmospheric air, (2) chill-stored pre-frozen pork chops packed in atmospheric air, and (3) freezer-stored, vacuum-packed pre-cooked sausages was investigated. The addition of 6% rapeseed oil did not influence the oxidative stability of the pork chops negatively, whereas the oxidative stability of a product such as the pre-cooked sausages (Danish dinner sausages) with a higher fat content (15%) decreased as a result of rapeseed oil feeding. Inclusion of rapeseed oil in the diets increased the amount of monounsaturated fatty acids and polyunsaturated fatty acids (PUFAs) in the meat and fat for the production of sausages at the expense of the content of the saturated fatty acids, and the higher content of PUFAs readily explains the decreased oxidative stability of the pre-cooked sausages. Feeding pigs 100 mg or 200 mg all-rac-α-tocopheryl acetate/kg feed significantly increased the oxidative stability of the pork chops and the detrimental effect of rapeseed oil observed in the pre-cooked sausages was effectively neutralised by both levels of vitamin E supplementation. Supplementation with copper did not affect the oxidative stability of any of the products. The presented results show that it is possible to produce pork products with a nutritionally improved fatty acid profile by inclusion of 6% rapeseed oil, without affecting the oxidative stability of the products negatively, through the protection provided by dietary vitamin E. Received: 23 March 1998 / Revised version: 28 May 1998     

Utilisation of pectin coating to enhance spray-dry stability of pea protein-stabilised oil-in-water emulsions

In this study, development of pea (Pisum sativum) protein stabilised dry and reconstituted emulsions is presented. Dry emulsions were prepared by spray-drying liquid emulsions in a laboratory spray-dryer. The effect of drying on the physical stability of oil-in-water emulsions containing pea protein-coated and pea protein/pectin-coated oil droplets has been studied. Oil-in-water emulsions (5 wt.% Miglyol 812 N, 0.25 wt.% pea protein, 11% maltodextrin, pH 2.4) were prepared that contained 0 (primary emulsion) or 0.2 wt.% pectin (secondary emulsion). The emulsions were then subjected to spray-drying and reconstitution (pH 2.4). The stability of the emulsions to dry processing was then analysed using oil droplet size, microstructure, Zeta potential, and creaming measurements. Obtained results showed that the secondary emulsions had better stability to droplet aggregation after drying than primary emulsions. To interpret these results, we propound that pectin, an anionic polysaccharide, formed a less charged protective layer around the protein interfacial film surrounding the oil droplets that improved their stability to spray-drying mainly by increasing steric effects.     

Effect of Antioxidants and Cooking on Stability of n-3 Fatty Acids in Fortified Meat Products

ABSTRACT: This study was carried out to determine the effect of processing and cooking on n-3 polyunsaturated fatty acid (PUFA) stability and to determine the efficacy of antioxidants (ANTI) to minimize lipid oxidation in cooked n-3 PUFA-fortified meat products. An emulsion of n-3 PUFAs (25% algal oil) was incorporated into ground turkey, pork sausages, or restructured hams (500 mg n-3 PUFA/110 g meat) with or without an "antioxidant cocktail" containing citrate (0.5% w/w), erythorbate (1 g/kg product), and rosemary (0.2% w/w). Ground turkey and pork sausages were frozen 2 d, then cooked to 71°C, and stored at 4°C for 2 d. Cooked, restructured hams were sliced, vacuum packaged, and stored at 4°C, or frozen and thawed with subsequent storage at 4°C. Treatments were CON (control), n-3 (n-3 PUFAs), CON + ANTI and n-3 + ANTI. Products were analyzed for color, lipid oxidation (TBARS and peroxide values), and n-3 PUFA profile. TBARS of n-3 PUFA-fortified treatments in ground turkey and pork sausages increased with storage ( P < 0.05); there were no changes in TBARS for CON + ANTI and n-3 + ANTI groups ( P > 0.05). For restructured hams nitrite curing appeared to delay lipid oxidation such that antioxidant treatment effects were unobservable ( P > 0.05). Overall recovery of n-3 PUFAs after heat processing was 69% to 85%, and there was no effect of storage on n-3 PUFA concentration in raw or cooked products ( P > 0.05). Sensory scores for n-3 treated restructured hams were lower than controls ( P < 0.05). Results suggested that cooking resulted in some losses of n-3 PUFAs in fortified meat products and that an "antioxidant cocktail" protected against lipid oxidation during subsequent storage in non-cured meat products.     

Quality characteristics of Dutch-style fermented sausages manufactured with partial replacement of pork back-fat with pure, pre-emulsified or encapsulated fish oil

Dutch-style fermented sausages were manufactured with 15% and 30% pork back-fat substitution by pure or commercial encapsulated fish oil, either added as such or as pre-emulsified mixture with soy protein isolate. Adding commercial encapsulated fish oil was the most important factor influencing the chemical composition. The fat content was not significantly different between products (p > 0.05). The n−6/n−3 ratio decreased from 8.49 in controls to 0.90-2.47 in modified products. Lipid oxidation parameters (propanal and hexanal) showed much higher values for sausages with pure fish oil than for products with encapsulated oil. For the latter, lipid oxidation was similar to controls. Products with encapsulated or pre-emulsified oil were significantly firmer than products from other treatments in physical and sensory analysis (p < 0.05). Overall, it is technologically feasible to enrich dry fermented sausages with n−3 fatty acids from fish oil and the application of commercial encapsulated fish oil seems to be the best in retaining overall quality.     

Chemical and physical stability of protein and gum arabic-stabilized oil-in-water emulsions containing limonene

ABSTRACT:  An important flavor component of citrus oils is limonene. Since limonene is lipid soluble, it is often added to foods as an oil-in-water emulsion. However, limonene-containing oil-in-water emulsions are susceptible to both physical instability and oxidative degradation, leading to loss of aroma and formation of off-flavors. Proteins have been found to produce both oxidatively and physically stable emulsions containing triacylglycerols. The objective of this research was to determine if whey protein isolate (WPI) could protect limonene in oil-in-water emulsion droplets more effectively than gum arabic (GA). Limonene degradation and formation of the limonene oxidation products, limonene oxide and carvone, were less in the WPI- than GA-stabilized emulsions at both pHs 3.0 and 7.0. These data suggest that WPI was able to inhibit the oxidative deterioration of limonene in oil-in-water emulsions. The ability of WPI to decrease oxidative reactions could be due to the formation of a cationic emulsion droplet interface at pH 3.0, which can repel prooxidative metals, and/or the ability of amino acids in WPI to scavenge free radical and chelate prooxidative metals.     

Stability of linseed oil and antioxidants containing dry fermented sausages: A study of the lipid fraction during different storage conditions

Different packaging conditions (aerobic, vacuum and modified atmosphere) were evaluated in order to study the stability of the lipid fraction of dry fermented sausages manufactured with a partial substitution of pork backfat by linseed oil and antioxidants. After 5 months of storage, α-linolenic acid was better preserved by vacuum and MAP (7.32 and 7.74g/100g fatty acids, respectively) than in aerobic conditions (6.15g/100g fatty acids), without significant differences to values obtained after 2 months of storage for this acid. At the end of the storage, (n-6)/(n-3) fraction in sausages with linseed oil was in all cases lower than 3, in contrast to values obtained for control products that were all higher than 15. Better PUFA/SFA ratios were also observed in modified sausages (0.6-0.7g/100g fatty acids) than control ones (0.3-0.4g/100g fatty acids). No signs of lipid oxidation measured by TBARs and peroxides were detected for modified sausages regardless the packaging system used (TBARs values lower than 0.25ppm and peroxides lower than 4meq O(2)/kg), pointing at a high effectiveness of the antioxidants. Furthermore, vacuum and MAP prevented 2,4-decadienal formation. Nutritional benefits of linseed oil and antioxidants containing products were maintained after 5 months of storage.     

Impact of free fatty acid concentration and structure on lipid oxidation in oil-in-water emulsions

Free fatty acids are strong prooxidants in both bulk and emulsified oils. Addition of oleic acid to an oil-in-water emulsions increased lipid hydroperoxide and hexanal formation at free fatty acid concentrations as low as 0.1% of the lipid. The prooxidant effect of free fatty acids was dependent on fatty acid type with lipid oxidation rates being in the order of linolenic < linoleic < oleic. There were no significant differences in lipid oxidation rates when free fatty acid isomers with cis or trans double bonds were compared. The prooxidant activity of the free fatty acids was postulated to be due to their ability to attract prooxidant metals as well as co-oxidise the triacylglycerol in the oil. Overall, these results show that the oxidative stability of oil-in-water emulsions is strongly linked to both the concentration and type of free fatty acids present.     

Changes of fatty acid composition during the processing of fish

Since the 70's a special attention has been paid to fish oil for its beneficial nutritive effects [1]. Fish products are an important source of polyunsaturated fatty acids (PUFAs) especially n-3 (ω-3) fatty acids - eicosapentaenoic (20:5, n-3) and docosahexaenoic (22:6, n-3) acid which are believed to cause the lowering of the incidence of thrombosis and atherosclerosis [2]. Due to the chemical properties all PUFAs undergo oxidative changes yielding products without any positive nutrition effects. Therefore, the processed fish - preserved, marinated or smoked - differ in the n-3 PUFAs content from the fresh fish meat. The present work aims to compare the effect of smoking and marinating on the n-3 fatty acids content of mackerel meat.     

Enriching breakfast sausages by feeding pigs with CLA supplemented diets

Approaches for improving the profile of functional unsaturated fatty acids in pork products include dietary supplementation of pigs with functional oils. Little information is available to indicate the benefit of this approach in a processed and cooked pork product such as breakfast sausages. Therefore, the aim of the present study is to examine the fatty acid profile and oxidation level in cooked pork sausages, produced following dietary supplementation with CLA compared to sunflower oil (SFO). Fat and moisture percentages, total fatty acid profiles and TBARS were analysed. Fatty acid profiles were altered in the sausages following all treatments. While a stronger effect was seen for CLA treatments, addition of SFO in the diet also resulted in linear increases of CLA in the sausages. CLA supplementation resulted in increased saturated fatty acid content; however, all treatments were within the recommended polyunsaturated/saturated fatty acid ratio of above 0.4. Improved oxidative stability was observed in sausages from CLA supplemented diets.     

LIPID OXIDATION AND SENSORY CHARACTERISTICS OF FRESH AND CURED SAUSAGES FROM α-LINOLENIC ACID ENRICHED PORK

Forty-eight female piglets were raised with four levels of α-linolenic acid (0, 1.5, 2.5, 3.5%) to determine the effect of dietary fat on the fatty acid composition, sensory and storage stability of pork products. The degree of polyunsaturation in neutral lipids and phospholipids were increased significantly by the dietary LNA. The thiobarbituric acid reactive substances (TBARS) vaues of freshly prepared breakfast sausages with vacuum packaging remained low, but those of the loosely packaged, i. e., nonvacuumized, sausages increased more than 3 times during the 48 h storage period. The TBARS values of breakfast sausages frozen-stored for 3 months showed extensive lipid oxidation. The dietary LNA treatments and packaging had significant effects on the susceptibility of breakfast sausages to lipid oxidation. However, the oxidative stability of cured products was not affected by the dietary treatments. The results indicated that the pork enriched with polyunsaturated fatty acids could be used in cured products or uncured meat products protected by ‘hot vacuum packaging’without any sensory or quality deterioration due to lipid oxidation.     

The interplay between diverse oil body extracts and exogenous biopolymers or surfactants

Hazelnuts, sesame seeds and soybeans were selected as three diverse sources of oil bodies. Application of aqueous extraction and centrifugation steps resulted in concentrated oil body creams that were studied for their physical stability after dilution to a series of 5.0 wt.% oil-in-water emulsions incorporating sodium caseinate (1.0 wt.%), Tween 80 (1.0 wt.%) or xanthan gum (0.1 wt.%). In terms of aggregation/coalescence and creaming, the stability of the oil body based emulsions was ruled to a large extent by the initial natural oil droplet size and the presence of co-extracted exogenous proteins and secondarily by the added biopolymers and the surfactant. More specifically, soybean oil bodies exhibited the highest physical stability, even though incorporation of Tween 80 into all three oil body emulsions improved the stability against aggregation/coalescence, while xanthan gum was an effective stabilizer against creaming.     

Healthy reduced-fat Bologna sausages enriched in ALA and DHA and stabilized with Melissa officinalis extract

Reduced-energy and reduced-fat Bologna products enriched with ω-3 polyunsaturated fatty acids were formulated by replacing the pork back-fat by an oil-in-water emulsion containing a mixture of linseed-algae oil stabilized with a lyophilized Melissa officinalis extract. Healthier composition and lipid profile was obtained: 85 kcal/100 g, 3.6% fat, 0.6 g ALA and 0.44 g DHA per 100 g of product and ω-6/ω-3 ratio of 0.4. Technological and sensory problems were not detected in the new formulations. Reformulation did not cause oxidation problems during 32 days of storage under refrigeration. The results suggest that it is possible to obtain reduced-fat Bologna-type sausages rich in ALA and DHA and stabilized with natural antioxidants, applying the appropriate technology without significant effects on the sensory quality, yielding interesting products from a nutritional point of view.     

Influence of interfacial composition on oxidative stability of oil-in-water emulsions stabilized by biopolymer emulsifiers

The objective of this research was to evaluate the influence of storage pH (3 and 7) and biopolymer emulsifier type (Whey protein isolate (WPI), Modified starch (MS) and Gum arabic (GA)) on the physical and oxidative stability of rice bran oil-in-water emulsions. All three emulsifiers formed small emulsion droplets (d₃₂ < 0.5 μm) when used at sufficiently high levels: 0.45%, 1% and 10% for WPI, MS and GA, respectively. The droplets were relatively stable to droplet growth throughout storage (d₃₂ < 0.6 μm after 20 days), although there was some evidence of droplet aggregation particularly in the MS-stabilized emulsions. The electrical charge on the biopolymer-coated lipid droplets depended on pH and biopolymer type: −13 and −27 mV at pH 3 and 7 for GA; −2 and −3 mV at pH 3 and 7 for MS; +37 and −38 mV at pH 3 and 7 for WPI. The oxidative stability of the emulsions was monitored by measuring peroxide (primary products) and hexanal (secondary products) formation during storage at 37 °C, for up to 20 days, in the presence of a pro-oxidant (iron/EDTA). Rice bran oil emulsions containing MS- and WPI-coated lipid droplets were relatively stable to lipid oxidation, but those containing GA-coated droplets were highly unstable to oxidation at both pH 3 and 7. The results are interpreted in terms of the impact of the electrical characteristics of the biopolymers on the ability of cationic iron ions to interact with emulsified lipids. These results have important implications for utilizing rice bran oil, and other oxidatively unstable oils, in commercial food and beverage products.     

Effect of dietary fish oil on fatty acid composition, lipid oxidation and sensory property of chicken frankfurters during storage

Broilers fed with three levels of fish oil (0, 2 and 4%) for 6 weeks were used in the manufacture of chicken frankfurters. The meat samples were vacuum-packed and stored at 0°C for 0, 10, 20 and 30 days. Cooking yield, proximate and fatty acid composition, lipid oxidation, microbial and sensory properties of the meat products were measured. Cooking yield, moisture, fat, protein, ash and cholesterol contents of frankfurters were not affected (P>0.05) by levels of dietary fish oil. Frankfurters processed from chickens fed diets enriched with 2 and 4% of supplemental fish oil had a higher (P<0.05) level of n-3 fatty acids (eicosapentaenoic acid and docosahexaenoic acid), but had a lower level of n-6 fatty acids (P<0.05) than the controls (0% fish oil). TBA values and pH values of meat samples were not affected (P>0.05) by levels of supplemental fish oil in the diets, however, pH values of the vacuum-packaged chicken frankfurters decreased (P<0.05) as the storage time increased. No differences (P>0.05) in total anaerobic plate counts were observed among the fish oil treatments during storage. Supplementation of 2 and 4% fish oil in the diets did not increase (P>0.05) the intensity of fishy flavor in chicken frankfurters.     

Development of combinations of chemically modified vegetable oils as pork backfat substitutes in sausages formulation

Today’s consumers look for foods which provide nutrition and pleasure, while safeguarding their health, the result of which is that they increasingly avoid foods containing cholesterol and saturated and trans fatty acids. Chemically modified vegetable oils can help tailor meat products to meet this growing need and at the same time fulfil the technological needs of the meat processing industry. In this study, 16 backfat samples were characterised for their solid fat content (SFC) and melting point and these characteristics were used to design a mixture of chemically modified vegetable oils for use as a pork fat substitute for elaborating sausages. The mixtures were prepared with different vegetable oils bearing in mind with stearic acid content due to its close correlation with the SFC. The backfat was characterised as a function of its SFC and some modified vegetable oil mixtures were proposed, which led to a 10–20% diminution in saturated fatty acids and with a melting point similar to those observed in the backfat. The fatty acid profile pointed to a polyunsaturated/saturated fatty acids ratio higher than 0.4, and an n − 6/n − 3 fatty acid ratio of less than 4 in both modified vegetable oil mixtures proposed.     

富含n-3LC-PUFA调和油的家庭烹饪氧化稳定性研究

通过对DHA和EPA质量分数分别为0.23%、0.45%、0.90%、1.80%的调和油进行家庭炒菜试验的研究,考察了富含n-3长碳链多不饱和脂肪酸(Long chain polyunsaturated fatty acid,LC-PUFA)的调和油的家庭烹饪氧化稳定性。结果表明:炒土豆丝后,上述油样中DHA和EPA的损失分别为1.2%、2.9%、2.4%、1.4%;炒青椒肉片后其损失分别为:3.0%、2.2%、2.4%、4.5%。炒菜前后调和油的各项指标包括羰基价、总氧化值(TOTOX)、硫代巴比妥酸值(TBA)、酸价和聚合物含量与大豆油(对照样1)以及不含DHA和EPA的调和油(对照样2)相比,油样的氧化程度以及油脂品质与两个对照样接近。不同DHA和EPA含量的调和油炒菜之后其多不饱和脂肪酸保留率在95%以上,氧化稳定性和豆油接近,适合家庭烹饪。     

Crosslinking of interfacial layers in multilayered oil-in-water emulsions using laccase: Characterization and pH-stability

The enzymatic crosslinking of polymer layers adsorbed at the interface of oil-in-water emulsions was investigated. A sequential two step process, based on the electrostatic deposition of pectin onto a fish gelatin interfacial membrane was used to prepare emulsions containing oil droplets stabilized by fish gelatin-beet pectin membranes (citrate buffer, 10 mM, pH 3.5). First, a fine dispersed primary emulsion (5% soybean oil (w/v), 1% (w/w) gelatin solution) (citrate buffer, 10 mM, pH 3.5) was produced using a high pressure homogenizer. Second, a series of secondary emulsions were formed by diluting the primary emulsion into pectin solutions (0 - 0.4% (w/w)) to coat the droplets. Oil droplets of stable emulsions with different oil droplet concentrations (0.1%, 0.5%, 1.0% (w/v)) were subjected to enzymatic crosslinking. Laccase was added to the fish gelatin-beet pectin emulsions and emulsions were incubated for 15 min at room temperature. The pH- and storage stability of primary, secondary and secondary, laccase-treated emulsions was determined. Results indicated that crosslinking occurred exclusively in the layers and not between droplets, since no aggregates were formed. Droplet size increased from 350 to 400 nm regardless of oil droplet concentrations within a matter of minutes after addition of laccase suggesting formation of covalent bonds between pectin adsorbed at interfaces and pectin in the aqueous phase in the vicinity of droplets. During storage, size of enzymatically treated emulsions decreased, which was found to be due to enzymatic hydrolysis. Results suggest that biopolymer-crosslinking enzymes could be used to enhance stability of multilayered emulsions.