Production of n − 3 polyunsaturated fatty acid concentrate from sardine oil by lipase-catalyzed hydrolysis

The production of n − 3 polyunsaturated fatty acids (n − 3 PUFAs) concentrate from oil extracted from Pacific sardines (Sardinops sagax) was studied using lipase-catalyzed hydrolysis. Commercially available microbial lipases, from Candida Rugosa (CR), Candida cylindracea (CC), Mucor javanicus (MJ), and Aspergillus niger (AN) were used for enzymatic hydrolyses with extracted sardine oil, run at 37 °C with constant stirring for 1.5, 3, 6, and 9 h. Fatty acid composition analysis by gas chromatography showed that the refined unhydrolyzed oil contained 26.86% of eicosapentaenoic acid (EPA) and 13.62% of docosahexaenoic acid (DHA) (wt/wt%). CR lipase was the most effective in concentrating n − 3 PUFA. Hydrolysis with 250 U CR lipase increased EPA concentration to a relatively constant level of 33.74% after 1.5 h. DHA levels were also significantly increased from 13.62% to 29.94% with 500 U after 9 h. Compared to CR and CC lipases, MJ and AN lipases resulted in low n − 3 PUFA concentration. Triacylglycerol levels decreased significantly as reaction time progressed.     

Nutritional and sensory aspects of light lamb meat enriched in n−3 fatty acids during refrigerated storage

Fatty acid content and sensory characteristics of meat from light lambs fed three diets supplemented with different sources of n−3 fatty acids (fish oil, extruded linseed and extruded linseed plus microalgae) and a control diet during refrigerated storage have been studied. The meat from lambs fed linseed diets had the highest levels of C18:3 n−3, while animals fed fish oil had the highest long-chain n−3 polyunsaturated fatty acids (PUFA). Thus, 100 g of meat from lamb fed the fish oil diet provided 183 mg of long-chain n−3 PUFA, representing 40% of the daily recommended intake. The levels of n−3, n−6 and long-chain n−3 PUFA decreased during a 7-day storage period. The meat from lambs fed the fish oil diet had high scores of fish odour and flavour and rancid odour and flavour and the lowest overall liking. Rancid odour and flavour increased after storage, mainly in supplemented groups.     

Evaluation of lipid oxidation in spaghetti pasta enriched with long chain n−3 polyunsaturated fatty acids under different storage conditions

This study investigated lipid oxidation in spaghetti enriched in long chain n−3 polyunsaturated fatty acids (LC n−3 PUFA) by the addition to semolina of an integrator containing eicosapentaenoic acid and docosahexaenoic acid. Two oxidative parameters were evaluated: peroxide value (PV), to assess primary oxidation and oxidised fatty acids to quantify secondary oxidation products. Functional spaghetti had a shelf life comparable to control pasta. LC n−3 PUFA were not significantly implicated in the onset of oxidation in spaghetti stored under daylight and accelerated oxidation in a laboratory heater. Storage decidedly affected shelf life: PV in functional spaghetti increased from 7.1 to 43.4 meq O₂/kg of fat under light exposure over 12 months, and from 7.1 to 16.2 meq O₂/kg under accelerated ageing at 55 °C for 27 days, reproducing about 18 months at room temperature. Oxidised fatty acids increased in fortified spaghetti from 4.8 to 13.8 g/100 g of fat under light exposure over 12 months and from 4.8 to 7.8 g/100 g of fat at 55 °C in 27 days. The high sensitivity of spectrophotometric and chromatographic analytical methods permitted the evaluation of primary and secondary oxidative derivatives in small amounts of fat.     

Fatty acids and sensory characteristics of Spanish dry-cured loin enriched in acid α-linolenic and α-tocopherol

The effects of using α-linolenic and α-tocopherol acid-enriched pork on the fatty acids and sensory characteristics of Spanish dry-cured loins were investigated. For the study, five batches of Spanish dry-cured loins were manufactured using pork loin enriched in polyunsaturated n − 3 fatty acids and α-tocopherol. Tissues were obtained from pigs fed on diets with the same ingredients, except for the oil source which corresponded to: [sunflower (C), linseed (L) and linseed and olive (1/1, w/w) (LO)] and two different amounts of α-tocopheryl acetate [20 (C, L and LO) or 200 (LOE and LE) mg/kg diet]. Dry-cured loins with polyunsaturated fatty acid n6/n3 ratios below 4 were obtained from linseed and linseed/olive oil-enriched batches. Dry-cured loin manufactured with pork from animals fed on diets enriched only with linseed oil presented the worst sensory characteristics and higher TBAR values than did dry-cured loins from animals fed on diets enriched with linseed and olive oil and linseed oil plus tocopheryl acetate.     

Physicochemical changes in ω − 3-enhanced farmed rainbow trout (Oncorhynchus mykiss) muscle during refrigerated storage

Physicochemical changes of ω − 3-enhanced farmed rainbow trout (Oncorhynchus mykiss) fillets developed by dietary modification with flaxseed oil and α-tocopheryl acetate (α-TA) were determined during storage at 2 °C. Trout were fed experimental diets for 120 days followed by processing to obtain boneless skinless fillets. The dietary modification increased concentration of total ω − 3 fatty acids in the fillets, which enhanced chances for lipid oxidation during storage. The fillets were vacuum or non-vacuum packed and stored at 2 °C for 10 or 12 days. Dietary α-TA resulted in higher (P < 0.05) concentration of α-tocopherol in fillets during storage; however, it did not retard (P > 0.05) lipid oxidation. Vacuum packaging resulted in much lower (P < 0.05) TBARS and higher (P < 0.05) retention of α-tocopherol during storage than non-vacuum packaging. However, α-tocopherol unlike vacuum packaging better protected ω − 3 FA in the fillets during storage.     

Enrichment of eggs in n-3 polyunsaturated fatty acids by feeding hens with different amount of linseed oil in diet

The production of n-3 polyunsaturated fatty acid enriched eggs by addition of linseed oil to the laying hens' diet has been evaluated in terms of production parameters and n-6/n-3 ratio. A total of 150 18weeks old Lohmann Brown laying hens were housed in cages and fed with basal diet and four experimental diets containing 1%, 2%, 3% or 4% of linseed oil added to the basal diet. The effect of the altered level of linseed oil on hens laying performance, fatty acid content and composition and cholesterol content in egg yolk has been evaluated during 13weeks of experiment. Egg weight, yolk fat content, yolk weight, yolk percentage and shape index were not influenced by dietary treatment. The ratio between n-6 and n-3 polyunsaturated fatty acids in eggs decreased in first 5weeks and then remained stable until the end of the experiment for all experimental groups. Different contents of linseed oil in feed highly influenced the n-6/n-3 ratio (P<0.0001). Addition of linseed oil did not influence the cholesterol content in yolks (P=0.5200) while the only factor affecting the cholesterol content was the hens age (P<0.0001).     

Healthier oils stabilized in konjac matrix as fat replacers in n − 3 PUFA enriched frankfurters

Nutritional, sensory and technological properties of frankfurters as affected by reformulation processes designed to reduce fat content and improve fatty acid profile were investigated. Healthier oils stabilized in oil in water emulsion or in konjac matrix gel were used as fat replacers. Results showed that improved fat content by the replacement of pork backfat with konjac gel and by the addition of healthier oils stabilized by various different systems, both resulted in products with very similar characteristics. From a nutritional standpoint, reformulated frankfurters with konjac gel and/or added a healthier oil combination may claim “reduced fat content” and/or “high omega 3 fatty acid content” according to European Regulation, since they could contain less than 30% of the fat in the reference product and more than 0.6 g of ALA/100 g and more than 80 mg of the sum of EPA plus DHA per 100 g, respectively. Chill storage over 40 days generally had little effect on the technological characteristics of frankfurters.     

Seasonal variation in the fatty acid composition of three Mediterranean fish – sardine (Sardina pilchardus), anchovy (Engraulis encrasicholus) and picarel (Spicara smaris)

The total fat contents and the fatty acid compositions of three common Mediterranean fish, namely sardine (Sardina pilchardus), anchovy (Engraulis encrasicholus) and picarel (Spicara smaris) were determined at bimonthly intervals for a one-year period. The purpose of this work was to study the seasonal variation of the fatty acids in the three fish that are some of the best sources of n − 3 fatty acids. The fat and fatty acid content of the investigated fish species show a significant seasonal dependency. Two of the fish (anchovy and picarel) have the highest fat content during the late winter – spring period. On the other hand, sardine shows the highest fat concentrations during the spring-early summer period. The fish that showed the highest variation in fatty acid composition was the anchovy. The sardine was found to be the best source of n − 3 fatty acids during the one-year period (35.35 g/100 g fatty acids). Finally the picarel had the highest oleic acid content (on average, 13.89/100 g fatty acids).     

Fishmeal-based diet decreases the redness of sunshine bass (Morone chrysops × Morone saxatilis) fillets

The objectives of the present study were to determine the effects of feeding a fishmeal-based diet on color attributes and lipid oxidation in sunshine bass (Morone chrysops × Morone saxatilis) fillets during retail display. A balanced diet containing 30 percent fishmeal (FM) or a diet containing poultry byproduct meal as a complete replacement of fishmeal (PB) was fed to sunshine bass for fifteen months. Harvested fish were filleted, overwrapped with polyvinyl chloride film and stored at 2 °C (REF) or over ice (ICE), under an illuminated retail display. Samples (n = 6) were analyzed after 0, 3, 6, or 9 d storage for color attributes (CIE L^∗, a^∗, b^∗, hue angle and chroma), thiobarbituric acid-reactive substances (TBARS), and pH. TBARS and pH increased (P < 0.05) during storage, indicating progress in lipid oxidation and protein changes. FM fillets demonstrated lower (P < 0.05) a^∗ (redness) value and greater (P < 0.05) hue angle than PB fillets. Since consumer acceptance of sunshine bass is dependant upon its white flesh, fishmeal supplementation could be used as a dietary strategy to improve fish marketability.     

A comparative analysis of the fatty acid profiles in the liver and muscles of male and female Salmo trutta macrostigma

The fatty acid compositions of liver and muscle of male and female Salmo trutta macrostigma, in the Tohma River, Turkey, were determined by gas chromatography. There were quantitative differences between individual fatty acids in the tissues investigated, depending on the sex. The most abundant fatty acids in both tissues of both sexes were palmitic acid (C16:0; 19.0–21.6%), stearic acid (C18:0; 5.32–11.3%), C18:1 ω−7 (5.65–9.38%), oleic acid (C18:1 ω−9; 15.6–22.4%), eicosapentaenoic acid (EPA; C20:5 ω−3; 6.34–7.88%) and docosahexaenoic acid (DHA; C22:6 ω−3; 7.38–15.6%). The ω−3/ω−6 ratio in tissues were found to be 2.89 (♂) and 1.97 (♀) in liver, and 2.59 (♂) and 2.26 (♀) in muscle. S. trutta macrostigma may be a valuable food for human consumption in terms of fatty acids.     

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.     

Effects of feeding flaxseed or sunflower-seed in high-forage diets on beef production,quality and fatty acid composition

Yearling steers were fed 70:30 forage:concentrate diets for 205 d, with either grass hay (GH) or red clover silage (RC) as the forage source, and concentrates containing either sunflower-seed (SS) or flaxseed (FS), each providing 5.4% oil to diets. Feeding diets containing SS versus FS significantly improved growth and carcass attributes (P < 0.05), significantly reduced meat off-flavor intensity (P < 0.05), and significantly increased intramuscular proportions of vaccenic (t11-18:1), rumenic (c9,t11-CLA) and n − 6 fatty acids (FA, P < 0.05). Feeding diets containing FS versus SS produced significantly darker and redder meat with greater proportions of atypical dienes (P < 0.05). A significant forage × oilseed type interaction (P < 0.05) was found for n − 3 FA, α-linolenic acid, and conjugated linolenic acid, with their greatest intramuscular proportions found when feeding the RC-FS diet. Feeding GH versus RC also significantly improved growth and carcass attributes, sensory tenderness (P < 0.05) and significantly influenced intramuscular FA composition (P < 0.05), but overall, forage effects on FA profiles were limited compared to effects of oilseed.     

The role of meat as a source of n-3 polyunsaturated fatty acids in the human diet

It is considered that consumption of very long chain (VLC, carbon chain length ?20) n − 3 PUFAs in most Western populations is sub-optimal and benefits in relation to chronic disease would be gained from increased consumption. This review examines the current contribution that meat makes to dietary intake of VLC n − 3 PUFA and given its current low contribution, how ruminant meat may be enriched. Enrichment both directly with VLC n − 3 fatty acids and indirectly by increasing intake by the animals of α-linolenic acid (ALNA; C18:3 n − 3) are considered. Since it now appears that dietary ALNA is a very limited source of VLC n − 3 PUFA in humans, the indirect route is controversial but since some forages are rich sources of ALNA this route has many sustainability and environmental attractions. Consideration is also given to the increased concentrations of trans and conjugated fatty acids that will arise from enriching ruminant meat with PUFA.     

Indoor fattening of lambs raised on pasture. Part 1: Influence of stall finishing duration on lipid classes and fatty acids in the longissimus thoracis muscle

Forty male Ile-de-France lambs (10 blocks of 4 homologous lambs) were used to study the effects of four feeding systems on muscle fatty acids (FA): raising and finishing on cool-season grasses (G), raising on the same grasses and stall-finishing, indoors, on concentrates and hay, respectively, for 22 (GSS) or 41 days (GSL), and stall-feeding indoors on concentrates and hay during both growing and finishing periods (S). Twenty-four lambs only (6 blocks) were retained for comparison of growth performances, lipid content in the longissimus thoracis muscle (LT) and their FA composition according to treatment. The 16 other lambs (4 blocks) were removed from the comparison, due to a large spread in the growth of the lambs towards the end of the trial. No significant effects of treatment were seen on the rate of growth (221, 228, 243 and 245±SE 8.0g/d, respectively, for G, GSS, GSL and S groups), and the lipid contents of the LT (2.22, 2.16, 2.17 and 2.52±SE 0.11g/100g fresh tissue). Grazing, lowered n-6 PUFA (polyunsaturated fatty acids), and increased n-3 PUFA and C18:2 c9t11 (conjugated linoleic acid cis9, trans11) compared to concentrate feeding. The main effects of grazing were not removed by a short period of finish indoors on concentrate (GSS group), but C20:4 n-6 and C22:6 n-3 contents achieved the lowest contents in this group, with significant differences from the values observed for GSL and S groups (C20:4 n-6) or from the three other groups (C22:6 n-3). After a longer period of finish on concentrate (GSL group), C18:3 n-3 (linolenic acid), C18:2 c9t11 and long chain (LC) n-3 PUFA were brought to the levels observed in the S group. In terms of adequacy for human health, the C18:2 n-6/C18:3 n-3 ratios were favourably low in the four groups (2.6, 3.6, 4.9 and 5.2±SE 0.7, respectively, for G, GSS, GSL and S groups), the level observed in the case of G group being significantly lower than for the three other groups and the level observed for GSS group being significantly lower than for the GSL and S groups.     

Lipid oxidation in n-3 fatty acid enriched Dutch style fermented sausages

Dutch style fermented sausages were manufactured with a substitution of 10%, 15% and 20% of pork backfat by flaxseed oil and canola oil, pre-emulsified with soy protein isolate. The 15% and 20% substitution were also reached by adding encapsulated flaxseed oil and encapsulated fish oil and by adding flaxseed oil, pre-emulsified with sodium caseinate, respectively. The products were sliced, packaged in an oxygen-enriched atmosphere and stored in the dark for 12 weeks at 7°C. No differences were detected in moisture, protein and fat content between control and modified sausages, with the exception of the formulation with sodium caseinate. The PUFA/SFA ratio increased from 0.30 in the control to 0.42-0.48 in the sausages with canola oil and to 0.49-0.71 in the sausages with flaxseed oil. The n-6/n-3 ratio decreased from 11.20 in the control to 6.94-5.12 in the sausages with canola oil and to 1.93-1.05 in the sausages with flaxseed oil. The addition of canola oil and encapsulated flaxseed oil resulted in a comparable shelf life as the control in terms of lipid oxidation. In the samples with addition of pre-emulsified flaxseed oil, especially with sodium caseinate, lipid oxidation clearly increased during storage. Physical and sensory analysis showed that the sausages with encapsulated fish oil and flaxseed oil resembled the control most.     

Nutritional composition of blubber and meat of hooded seal (Cystophora cristata) and harp seal (Phagophilus groenlandicus) from Greenland

Seal blubber and skin are widely used, but the utilisation of blubber and meat for human consumption is limited. The aim of this study was to evaluate the nutritional composition of seal blubber and meat. The fatty acid composition, selected minerals and trace-elements, vitamins, amino acids and proximal composition of blubber and meat from hooded seal (Cystophora cristata) and harp seal (Phagophilus groenlandicus) from the “West Ice” near Greenland were analysed. The results showed that seal blubber is an excellent source of long- and very long-chain (VLC) n − 3 polyunsaturated fatty acids (PUFAs), in addition to long- and VLC monounsaturated fatty acids (MUFAs). Eicosapentaenoic acid (EPA) content contributed to a clear separation between blubber and meat from the two species. The blubber of harp seal showed the highest EPA (9.2%), whereas the muscle of harp seal showed the lowest EPA (3%) content. Seal meat is lean with less than 2% total fat, mainly composed of MUFAs, long- and VLC n − 3 PUFAs. In addition, the meat contains a high amount of proteins with a well-balanced amino acid composition. The trace-element content of seal meat is very high, particularly iron (379 μg/g muscle in hooded seal) and zinc (30 μg/g muscle in harp seal), as also is the vitamin content, especially vitamins A, D₃ and B₁₂. The seals included in this study varied greatly in age and size, which in turn may be the principal reason for the great individual variation observed in nutritional composition. On average, however, consumption of only 40 g seal meat covers the recommended daily intakes of iron and vitamin B₁₂ for young women. In conclusion, as long as the products fulfil the amending legislations regarding contaminants, both seal blubber and meat, from the present species, represent high quality food regarding nutrients and bioactive components beneficial for human health.     

Development of dry fermented sausages rich in docosahexaenoic acid with oil from the microalgae Schizochytrium sp.: Influence on nutritional properties,sensorial quality and oxidation stability

Significant enrichment of dry fermented sausages in n−3 polyunsaturated fatty acids (PUFA) was achieved by incorporating algae oil from Schizochytrium sp. into an emulsion that partially substituted pork backfat from the traditional formulation. Two different levels of substitution were initially tested, both including BHA and BHT as antioxidants: (15% and 25%). A triangular test showed that products with 25% substitution were not sensorially acceptable. Sausages with 15% substitution supplied 1.30 g/100 g product of docosahexaenoic acid (DHA), with an interesting n−6/n−3 ratio of 2.62. No signs of oxidation were detected at the end of the ripening process, with low values of TBARs (<0.2 mg/kg), peroxides (<2 meq O₂/kg fat) and volatile aldehydes. Storage of these sausages under vacuum during 30 days totally guaranteed their stability, whereas after 90 days certain degree of oxidation was detected, only by means of aldehydes analysis. Hexanal, nonanal and dienals increased, whereas no increment in TBARs or peroxides were observed. Aerobical storage of these sausages was not viable due to a high oxidation susceptibility of the new formulation, despite the use of antioxidants.     

Acorns (Quercus rotundifolia Lam.) and grass as natural sources of antioxidants and fatty acids in the “montanera” feeding of Iberian pig: Intra- and inter-annual variations

In two consecutive years (2006/2007 and 2007/2008), fortnightly samples were taken to characterise the antioxidant composition and fatty acid profile of acorns and grass on which Iberian pigs feed during the period of “montanera” (free-range rearing system of the south-western of Iberian Peninsula). The antioxidant parameters analysed were: α- and γ-tocopherol, total phenolics compounds (TPC), lipophilic and hydrophilic antioxidant activities (LAA, HAA) (acorn and grass) and condensed and hydrolysable tannins (CT, HT) and protein precipitating capacity (PP) (acorns). To characterise the fatty acid profiles, the thirteen major fatty acids were assayed. For the acorns, it was found that there was intra-annual variability in all the antioxidants studied except α-tocopherol, and inter-annual variability in all except the protein precipitating capacity. The fatty acid profile also varied depending on the sampling date and the study year, especially in the saturated fatty acids (SFA) and C18:1 n-9 content. For the grass, there was intra- and inter-annual variability in all the antioxidant parameters studied, and in the proportions of the fatty acids C18:0, and C18:1. It could explain variations in the antioxidant and fatty acids composition of Iberian pig tissues from animals raised in different “montanera” seasons and even in the same “montanera” season.     

Enriching M. sternomandibularis with -tocopherol by dietary means does not protect against the lipid oxidation caused by high-pressure processing

We hypothesized that elevating the concentration of α-tocopherol in beef muscle tissue by dietary means would increase lipid stability following high-pressure processing. Beef M. sternomandibularis was obtained from cattle that had medium (4.92 μg/g) and high (7.30 μg/g) concentrations of α-tocopherol. Post-rigor, paired muscles samples were subjected to pressures of 0.1 (atmospheric), 200 or 800 MPa for 20 min at approximately 60 °C. Following high-pressure processing, measurements were made immediately (d 0) or on samples stored in the dark for 6 d at 4 °C (d 6). Intramuscular lipid was similar for each group (4.02% vs. 4.26%, respectively; P = 0.78), but lipid from the medium α-tocopherol muscle was more saturated and less monounsaturated than muscle from the high α-tocopherol group. High-pressure processing at 800 MPa and 60 °C did not reduce the amount of α-tocopherol but significantly reduced the concentration of linoleic acid (18:2n−6) in muscle from both production groups of cattle. Thiobarbituric acid reactive substances increased linearly with treatment pressure only in d 6 samples (day × pressure interaction P = 0.0001) and were higher overall (P = 0.02) in the high α-tocopherol muscle than in the medium α-tocopherol muscle. At d 6, lipid peroxides were decreased (P = 0.007) by high-pressure treatment and were higher (P < 0.0001) in the high α-tocopherol group than in the medium α-tocopherol group. Therefore, muscle from the high α-tocopherol cattle in this study had a greater accumulation of lipid peroxides by d 6, making the muscle from those cattle more susceptible to oxidation.     

A developed pre-column derivatization method for the determination of free fatty acids in edible oils by reversed-phase HPLC with fluorescence detection and its application to Lycium barbarum seed oil

Free fatty acids (FFAs) in edible oils are one of the most frequently determined quality indices during production, storage, and marketing. In this study, a simple, stable and sensitive method for the simultaneous determination of saturated and unsaturated FFAs from edible oils using 2-(11H-benzo[a]carbazol-11-yl)-ethyl-4-methylbenzenesulphonate (BCETS) as labelling reagent with fluorescence detection has been established. FFAs are derivatized by BCETS and separated on a reversed-phase Eclipse XDB-C₈ column with a gradient elution. The results indicated that all FFAs were found to be given an excellent linear response with correlation coefficients of >0.9994. The detection limits at a signal-to-noise ratio of 3 were in the range of 0.22–1.06 ng/mL. When applied to Lycium barbarum seed oils, the developed method showed good reproducibility. The effect of extraction methods including supercritical CO₂ and organic solvent extraction on the FFAs composition has also been investigated. Meanwhile, this method exhibits powerful potential for the trace analysis of short- and long-chain free fatty acids from edible oils, foodstuff and other complex samples.