FA composition of the oil extracted from farmed atlantic salmon (Salmo salar L.) viscera

The FA composition of visceral oil extracted from farmed Atlantic salmon (Salmo salar L.) viscera was studied. Seventeen FA were identified in the extracted visceral oil, and the major FA were 18∶1n9, 16∶0, 16∶1n7, 20∶5n3 (EPA), 14∶0, and 22∶6n3 (DHA). The percentages of saturated, monounsaturated, and polyunsaturated FA in the total FA were 31.7, 36.0, and 32.2%, respectively. Compared with other fish oils, oil from farmed Atlantic salmon had much higher EPA (1.64 g/100 g) and DHA (1.47 g/100 g) contents. The FA profile of the salmon visceral oil was similar to that of the salmon fillet. Thus, the salmon visceral oil could be a replacement for the oil obtained from edible salmon fillet and used in functional foods or feeds requiring a high level of omega-3 FA. Furthermore, producing visceral oil is also beneficial to salmon fish industry by adding value back to the processing waste.     

Characterization of farmed and wild salmon (Salmo salar) by a combined use of compositional and isotopic analyses

Gas chromatography, isotope ratio mass spectrometry, and high-resolution ²H site-specific natural isotope fractionation/nuclear magnetic resonance spectroscopy have been used to study the different kinds of fish oils and lipids extracted from muscle of wild and farmed salmon (Norway, Scotland). A statistical analysis of the fatty acid compositions, overall ²H and ¹³C isotope ratios, and molar fractions of the isotopomeric deuterium clusters was carried out to select the most efficient variables for distinguishing the different groups of salmons and fishes studied. A classification analysis based on four fatty acid compositions, three deuterium molar fractions, and the overall (D/H)_tot isotope ratio of fish oils completely assings the oils to the right group.     

Thermal and flow properties of oils from salmon heads

Thermal and flow properties of unrefined oils from the heads of red or pink salmon were evaluated. Major thermal degradation of the salmon oils occurred between 200 and 450°C. Red and pink salmon oils were completely decomposed at 533 and 668°C, respectively. The phase transition of salmon oils occurred over a wide range of temperatures. The melting points of −69.6 to −0.36°C and −64.7 to 20.8°C were observed for red and pink salmon oils, respectively. The enthalpy was 40 j/g for red salmon oil and 39 j/g for pink salmon oil. Specific heat capacity ranges of 0.8 to 1.6 and 1.3 to 2.3 j/g/°C were observed for red and pink salmon oils, respectively. Both salmon oils exhibited Newtonian flow behavior. Red salmon oil required higher magnitudes of energy (kj·mol⁻¹) to flow than pink salmon oil. The viscosity of salmon oils was temperature-dependent and could be predicted by the Arrhenius equation.     

Lipids and lipid antioxidant systems in developing eggs of salmon (Salmo salar)

Lipid class and fatty acid analyses were carried out on developing salmon eggs at four clearly defined pre-feeding stages, namely, fertilization, eyed egg stage (50 days), hatching (yolk sac fry, 98 days) and swim up fry (138 days). Measurements of components of the system considered to be involved in defense of cells against lipid peroxidation (glutathione peroxidase, EC 1.11.1.9, glutathione S-transferase, EC 2.5.1.18, reduced glutathione [GSH], α-tocopherol and ascorbic acid) were made at the same time. Levels of triacylglycerol decreased markedly during development, but there were few changes in fatty acid composition, indicating a non-selective utilization of fatty acids. Phosphatidylcholine was the dominant polar lipid (>94% by weight) in fertilized eggs. It was used preferentially during development so that in swim up fry the ratio phosphatidylcholine: phosphatidylethanolamine approached that found in fish muscle. Amounts of docosahexaenoic acid and arachidonic acid in polar lipids were significantly greater (p<0.01) in swim up fry than in fertilized eggs. Activities of the two enzymes were very low in the fertilized egg and remained low until hatching, when there was a concerted increase in their activity and in the concentration of GSH. Egg tocopherol concentrations decreased significantly during development, but whole body concentrations in swim up fry were not dissimilar from those in normal juvenile fish. Ascorbic acid, on the other hand, declined to very low levels in swim up fry; the restoration of this vitamin during first feeding seems vital to the well being of the fish.     

Angiotensin I-Converting Enzyme (ACE) Inhibitory Activity and ACE Inhibitory Peptides of Salmon (Salmo salar) Protein Hydrolysates Obtained by Human and Porcine Gastrointestinal Enzymes

The objectives of the present study were two-fold: first, to detect whether salmon protein fractions possess angiotensin I-converting enzyme (ACE) inhibitory properties and whether salmon proteins can release ACE inhibitory peptides during a sequential in vitro hydrolysis (with commercial porcine enzymes) and ex vivo digestion (with human gastrointestinal enzymes). Secondly, to evaluate the ACE inhibitory activity of generated hydrolysates. A two-step ex vivo and in vitro model digestion was performed to simulate the human digestion process. Salmon proteins were degraded more efficiently by porcine enzymes than by human gastrointestinal juices and sarcoplasmic proteins were digested/hydrolyzed more easily than myofibrillar proteins. The ex vivo digested myofibrillar and sarcoplasmic duodenal samples showed IC₅₀ values (concentration required to decrease the ACE activity by 50%) of 1.06 and 2.16 mg/mL, respectively. The in vitro hydrolyzed myofibrillar and sarcoplasmic samples showed IC₅₀ values of 0.91 and 1.04 mg/mL, respectively. Based on the results of in silico studies, it was possible to identify 9 peptides of the ex vivo hydrolysates and 7 peptides of the in vitro hydrolysates of salmon proteins of 11 selected peptides. In both types of salmon hydrolysates, ACE-inhibitory peptides IW, IY, TVY and VW were identified. In the in vitro salmon protein hydrolysates an ACE-inhibitory peptides VPW and VY were also detected, while ACE-inhibitory peptides ALPHA, IVY and IWHHT were identified in the hydrolysates generated with ex vivo digestion. In our studies, we documented ACE inhibitory in vitro effects of salmon protein hydrolysates obtained by human and as well as porcine gastrointestinal enzymes.     

The effect of enhanced intake of linoleic acid on the fatty acid composition of tissue polar lipids of post-smolt atlantic salmon (Salmo salar)

Duplicate groups of Atlantic salmon (Salmo salar) post smolts were given diets in which the lipid component was either fish oil or a mixture of corn oil and lard. This difference in the dietary lipid did not significantly affect growth over a period of sixteen weeks. Proportions of docosahexaenoic acid [22∶6(n−3)] and total (n−3) fatty acids in the polar lipids of liver and white muscle were unaffected by this difference in dietary lipid component over the time period used. Fish given the diet containing corn oil and lard had significantly higher levels of 20∶2(n−6), 20∶3(n−6) and 20∶4(n−6) in the polar lipids of these tissues than were present in the tissues of the fish given diets containing fish oil. There results suggest that linoleic acid [18∶2(n−6)] undergoes elongation and desaturation to arachidonic acid [20∶4(n−6)] in post-smolt Atlantic salmon.     

Variation with chain length in acute toxicity of alkylhydroxamic acids to salmon (Salmo salar) fry

The acute toxicity of saturatedn-alkylhydroxamic acids to salmon (Salmo salar) fry was determined. Within the seriesn-C₆H₁₃CONHOH ton-C₁₀H₂₁CONHOH, acute toxicity (indicated by death) increased with chain length. Shorter and longer chain compounds were not toxic under similar conditions. Factors governing the variation in toxicity with chain length and possible mechanisms of action are discussed. Presented in part at the AOCS Meeting, Atlantic City, October 1971.     

Regiospecificity profiles of storage and membrane lipids from the gill and muscle tissue of atlantic salmon (Salmo salar L.) grown at elevated temperature

Regiospecific and traditional analysis, of both storage and membrane lipids, was performed on gill, white muscle, and red muscle samples taken from Atlantic salmon (Salmo salar) to gauge the effect of elevated water temperature. The fish, fed a commercial diet, were held at an elevated water temperature of 19°C. Total n-3 PUFA, total PUFA, and n-3/n-6 and unsaturated/saturated fatty acid (UFA/SFA) ratios in the FA profile of the total lipid extract in the white muscle were fairly low compared with fish grown at 15°C. Adaptation of structural and storage lipids at elevated temperatures was shown by a significant (P<0.01) reduction in PUFA especially in the percentage of EPA (6–8%). Further adaptation was indicated by the percentages of SFA, which were significantly (P<0.05) higher in gill (56%) and white muscle (58%) polar lipid fractions and coincided with lower percentages of n-3, n-6, and total PUFA. The regiospecific profiles indicated a high affinity of DHA to the sn-2 position in both the TAG (61–68%) and polar lipid (35–60%) fractions. The combination of detailed regiospecific and lipid analyses demonstrated adaptation of cell membrane structure in Atlantic salmon grown at an elevated water temperature.     

Sensory characterization of pate from waste salmon (Salmo salar L.) products utilizing response surface methodology

The goal of the present study was the development of a salmon pate formulation with excellent nutritional and sensory properties, using by product components from de salmon industry. The optimized formulation was obtained using the response surface methodology. A 2n factorial and a composite central rotatable design was applied in the experience. Salt and xanthan hydrocolloid gum were the selected independent variables and sensory quality the response variable. Statistical analysis was utilized to estimate the fitted model. The optimum combination of selected experimental variables were 1.5% salt and 1.35% xanthan gum. Data from the chemical characterization of salmon pate showed an important protein content (20%) greater than liver pate (11.5%) and trout pate (14.7%). With regard to fat content, the concentration of this component in salmon pate was half the concentration found in liver pate. The shelf life of the optimized formula determined at 3 degrees C and 18 degrees C were 18 days and 8 days respectively.     

Fatty acyl desaturation in isolated hepatocytes from Atlantic salmon (Salmo salar): Stimulation by dietary borage oil containing γ-linolenic acid

The effects of different dietary oils on the fatty acid compositions of liver phospholipids and the desaturation and elongation of [1-¹⁴C]18∶3n−3 and [1-¹⁴C]18∶2n−6 were investigated in isolated hepatocytes from Atlantic salmon. Atlantic salmon smolts were fed diets containing either a standard fish oil (FO) as a control diet, a 1∶1 blend of Southern Hemisphere marine oil and tuna orbital oil (MO/TO), sunflower oil (SO), borage oil (BO), or oliver oil (OO) for 12 wk. The SO and BO diets significantly increased the percentages of 18:2n−6, 18:3n−6, 20:2n−6, 20:3n−6, and total n-6 polyunsaturated fatty acids (PUFA) in salmon liver lipids in comparison with the FO diet. The BO diet also increased the percentage of 20:4n−6. Both the SO and BO diets significantly reduced the percentages of all n−3 PUFA in comparison with the FO diet. The OO diet significantly increased the percentages of 18:1n−9, 18:2n−6, total monoenes, and total n−6 PUFA in liver lipids compared to the FO diet, and the percentages of all n−3 PUFA were significantly reduced. With [1-¹⁴C]18:3n−3, the recovery of radioactivity in the products of Δ6 desaturation was significantly greater in the hepatocytes from salmon fed SO, BO, and OO in comparison with the FO diet. The BO diet also increased the recovery of radioactivity in the products of Δ5 desaturation. Only the BO diet significantly affected the desaturation of [1-¹⁴C]18:2n−6, increasing recovery of radioactivity in both Δ6- and Δ5-desaturation products. In conclusion, dietary BO, enriched in γ-linolenic acid (18:3n−6), significantly increased the proportions of both 20:3n−6 and 20:4n−6 in salmon liver phospholipids and also significantly increased the desaturation of both 18:2n−6 and 18:3n−3 in salmon hepatocytes. The possible relationships between dietary fatty acid composition, tissue phospholipid fatty acid composition, and desaturation/elongation activities are discussed.     

Enrichment of salmon oil with n‐3 PUFA by lipolysis,filtration and enzymatic re‐esterification

Selective enzymatic hydrolysis of salmon oil extracted without solvent from by‐products was carried out under mild conditions, using a stereospecific sn‐1, sn‐3 lipase Novozyme^®. A modification of the lipid class composition was obtained by controlling the degree of hydrolysis (40%, 24 h). The mixture of acylglycerols and free fatty acids was submitted to a filtration step to retain in the retentate most of the saturated fatty acids, with melting peaks ranging from ‐31.9 °C to +14.7 °C obtained by differential scanning calorimetry. This step allowed a significant increase of polyunsaturated fatty acids (PUFA) from 39.2 mol‐% in the crude oil to 43.3% in the permeate. The remaining free fatty acids in the permeate (20.2 wt‐%) was re‐esterified with an immobilized 1, 3‐specific lipase IM60. Acylglycerols synthesis reached 90% in optimized conditions. After 48 h of reaction, the distribution of monoacylglycerols, diacylglycerols and triacylglycerols was 22.1, 28.7, 43.4 (w/w), respectively. The re‐esterification step did not modify the PUFA content obtained after membrane filtration.     

The effect of dietary lipid on polyunsaturated fatty acid metabolism in Atlantic salmon (Salmo salar) undergoing parr-smolt transformation

The aim of this study was to measure the changes in lipid metabolism which occur during smoltification and seawater transfer in Atlantic salmon (Salmo salar). Duplicate groups of Atlantic salmon parr were fed diets containing either fish oil (FO) or a blend of linseed and rapeseed oils, vegetable oil (VO), from October (week 0) to seawater transfer in May (week 26). From May to August (weeks 26–43), all fish were fed a fish oil-containing diet. Fatty acyl desaturation and elongation activity were followed in isolated hepatocytes incubated with radioactive 18:3n−3 and 18:2n−6. Metabolism of 18:3n−3 was consistently around 5-fold greater than metabolism of 18:2n−6, and total metabolism of both substrate polyunsaturated fatty acids (PUFA) was increased in fish fed both VO and FO up to seawater transfer after which desaturation activities were reduced. Desaturation activities with both 18:3n−3 and 18:2n−6 were significantly greater in fish fed VO, compared to fish fed FO, at 22 and 26 wk. Arachidonic acid (20:4n−6; AA) in liver polar lipids (PL) of fish fed VO increased consistently from weeks 0–22 but varied after seawater transfer. In fish fed FO, AA in liver PL remained constant up to week 17 before increasing at seawater transfer and leveling off thereafter. Eicosapentaenoic acid (20:5n−3; EPA) in liver PL of fish fed VO decreased significantly from week 0–22 before rising at seawater transfer and increasing rapidly posttransfer. EPA in liver PL of fish fed FO showed a similar trend except EPA was always greater in the freshwater phase compared to fish fed VO. Docosahexaenoic acid (DHA) levels in liver PL of fish fed VO remained constant in the freshwater phase before increasing following seawater transfer. In fish fed FO, DHA in liver PL increased from weeks 0–17 reducing and leveling off postseawater transfer. The levels of PGF_2α and PGF_3α were measured in isolated gill cells stimulated with calcium ionophore A23187. PGF_2α production in fish fed VO increased significantly between 0–7 wk before decreasing toward seawater transfer. After transfer, PGF_2α production increased to a peak at 35 wk. PGF_2α production in fish fed FO was not significantly altered during the trial period. The changes in PGF_3α production were broadly similar to those occurring with PGF_2α, but the latter was always in excess of the former (2-to 4-fold). Plasma chloride concentrations in fish subjected to seawater challenge at 20 wk were significantly lower in fish fed VO compared to those fed FO. This study has provided new information on the changes in lipid metabolism which accompany parr-smolt transformation and suggests that diets which have a fatty acid composition more similar to that in aquatic invertebrates may be beneficial in effecting successful seawater adaptation.     

Quantitative high-resolution13C and1H nuclear magnetic resonance of ω3 fatty acids from white muscle of atlantic salmon (Salmo salar)

High-resolution¹³C nuclear magnetic resonance (NMR) spectra have been obtained and used to define the ω3 (n-3) fatty acid distribution in lipid extract and white muscle from Atlantic salmon (Salmo salar). The¹³C spectrum of lipid extracted from muscle gives quantitative information about the individual n-3 fatty acids, 18:2n-6, 20:1/22:1 and groups of fatty acids. The quantitative data compare favorably with those obtained by gas-liquid chromatography. The¹H NMR spectrum of the lipid extract gives information about the amount of 22:6n-3 and the total content of n-3 fatty acids. The¹³C NMR technique also revealed the positional distribution (1,3- and 2-acyl) of the important 20:5n-3 and 22:6n-3 acids in the triacylglycerol molecules. In the quantitative¹³C NMR spectrum of white muscle, the methyl region of the acyl chains of triacylglycerols gave rise to sufficiently resolved signals to permit estimation of the total concentration of lipids and the n-3 fatty acid content. The NMR data are in good agreement with corresponding data obtained by traditional methods.     

Analysis of lipids from cooked beef by thin-layer chromatography with flame-ionization detection

Lipids were extracted from cooked ground beef with methylene chloride/methanol (2:1). The lipids were separated on a silicic acid column, into neutral and polar fractions by elution with methylene chloride, followed by methanol. These fractions were analyzed by Iatroscan thin-layer chromatography with flame-ionization detection instrumentation on Chromarods S-III (silica gelcoated quartz rods). Comparison of cooked beef stored for 0, 4 and 7 d at 4°C indicated that storage caused a decrease in total lipids, an increase in neutral lipids and a decrease in polar lipids, specifically in phosphatidylcholine. These changes in the lipid fraction were associated with meat flavor deterioration and an increase in lipid oxidation.     

Enrichment of phospholipids from neutral lipids in peanut oil by high-performance liquid chromatography

Phospholipids from crude peanut oil were enriched on a 2-cm silica column and subsequently separated from neutral lipids within the chromatographic system without prior concentration. Hexane effectively removed the bulk neutral lipids, leaving the adsorbed phospholipids on the silica precolumn. Individual phospholipids were separated from the remaining neutral lipids and from each other by using two mixed solvents and a gradient program. This method separates the phospholipids in approximately 27 min after the desired enrichment level has been reached. The research reported in this paper was a cooperative effort by the Agricultural Research Service of the United States Department of Agriculture and the North Carolina Agricultural Research Service, Raleigh, NC 27695-7625.     

The effect of dietary linoleic acid on the fatty acid composition of individual phospholipid and lipoxygenase products from gills and leucocytes of Atlantic salmon (Salmo salar)

Diets containing either fish oil or sunflower oil, both of which supplied the minimum required level of n−3 fatty acids, were given to Atlantic salmon (Salmo salar) postsmolts for a period of 16 weeks. In fish fed sunflower oil, the phospholipids of gills showed increased 18∶2n−6 (2–13-fold), 20∶2n−6 (4.5–12-fold) and 20∶−6 (2–8-fold). In addition, phosphatidylethanolamine had increased 20∶4n−6 (1.5-fold). Changes of a similar magnitude were observed in the phospholipids of blood leucocytes except that, in addition, 20∶4n−6 was elevated in phosphatidylserine (1.7-fold) and phosphatidylinositol (1.4-fold). Both tissues showed a general decrease in phospholipid 20∶5n−3 (up to 3-fold), which caused an increase in 20∶4n−6/20∶5n−3 ratio (1.3–6-fold). The elongation and desaturation products of 20∶4n−6, 22∶4n−6 and 22∶5n−6 were not increased as a result of feeding sunflower oil. When isolated gill cells were stimulated with the calcium ionophore A23187, 12-hydroxy-8,10,14,17-eicosapentaenoic acid (12-HEPE) was the major lipoxygenase product from salmon given fish oil. 12-HEPE was significantly reduced in salmon given sunflower oil. When stimulated with A23187, the lipoxygenase products derived from whole blood of fish given sunflower oil showed decreased levels of leukotriene B₅, 12-HEPE and 12-hydroxy-5,8,10,14-eicosatetraenoic acid.     

A Simplified Method to Distinguish Farmed (<Emphasis Type="Italic">Salmo salar</Emphasis>) from Wild Salmon: Fatty Acid Ratios Versus Astaxanthin Chiral Isomers

Mislabeling of farmed and wild salmon sold in markets has been reported. Since the fatty acid content of fish may influence human health and thus consumer behavior, a simplified method to identify wild and farmed salmon is necessary. Several studies have demonstrated differences in lipid profiles between farmed and wild salmon but no data exists validating these differences with government-approved methods to accurately identify the origin of these fish. Current methods are both expensive and complicated, using highly specialized equipment not commonly available. Therefore, we developed a testing protocol using gas chromatography (GC), to determine the origin of salmon using fatty acid profiles. We also compared the GC method with the currently approved FDA (United States Food and Drug Administration) technique that uses analysis of carotenoid optical isomers and found 100% agreement. Statistical validation (n = 30) was obtained showing elevated 18:2n-6 (z = 4.56; P = 0.0001) and decreased 20:1n-9 (z = 1.79; P = 0.07) in farmed samples. The method is suitable for wide adaptation because fatty acid methyl ester analysis is a well-established procedure in labs that conduct analysis of lipid composition and food constituents. GC analysis for determining the origin of North American salmon compared favorably with the astaxanthin isomer technique used by the FDA and showed that the fatty acid 18:2n-6 was the key indicator associated with the origin of these salmon.