Fractionation of squid visceral oil ethyl esters by short-path distillation

Squid visceral oil contains high levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Its ethyl esters were fractionated by short-path distillation in this study. The elimination temperatures of squid visceral oil ethyl esters (SVOEE) ranged from 50 to 140°C, increasing with the carbon number of ethyl esters. The elimination temperature of cholesterol was higher than those of SVOEE. The SVOEE of Illex argentinus (SVOEE-A) was more advantageous as the raw material (feed) than that of Ommastrephes bartrami (SVOEE-B) for the isolation of EPA and DHA, because SVOEE-A contained less 20∶1 and 22∶1. When SVOEE-A originally containing 9.0% EPA, 14.7% DHA, and 1,121 mg/100 g of cholesterol was distilled from 50 to 150°C with 20°C interval, the 130°C distillate could give 15.5% EPA and 34.7% DHA with 99 mg/100 g of cholesterol, and the yield was 21.8%. The 150°C distillate could give 43.1% DHA with 496 mg/100 g of cholesterol. Furthermore, the distillates collected from 110 to 150°C contained 24.4 to 50.2% of EPA plus DHA, and their total yield was 58.3%. The final residue after 150°C distillation contained 77% of the total cholesterol in the initial SVOEE-A, and the yield was 6.0%.     

Lipase‐catalysed enrichment of DHA and EPA in acylglycerols resulting from squid oil ethanolysis

The fatty acid specificity of four lipases towards eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) was evaluated when performing ethanolysis of squid oil. During the first part of ethanolysis, no DHA ethyl esters were detected when using the lipases from Thermomyces lanuginosus, Pseudomonas cepacia or Pseudomonas fluorescens (in the case of the second and third lipases, no EPA ethyl esters were detected either). This indicates that these three lipases could not catalyse the conversion of DHA located in a triacylglycerol to ethyl ester, and that the Pseudomonas lipases could not catalyse the conversion of EPA either. This pattern was not found for the lipase from Rhizomucor miehei. The lipase from Thermomyces lanuginosus showed the lowest specificity towards DHA and the highest DHA recovery during DHA enrichment in the acylglycerol fraction. It was thus used to catalyse the ethanolysis of squid oil on a larger scale. The ethyl esters formed were removed using short‐path distillation, resulting in a product containing mainly mono‐ and diacylglycerols. The product contained 34 mol‐% DHA and 17 mol‐% EPA, compared with 19 mol‐% DHA and 12 mol‐% EPA in the original squid oil.     

Combination of Urea Complexation and Molecular Distillation to Purify DHA and EPA from Sardine Oil Ethyl Esters

Urea complexation (UC) and the molecular distillation (MD) technique were applied jointly to purify eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from sardine oil ethyl esters (SOEE). Response surface methodology (RSM) was used to measure the influences of the variables to the responses and the optimal conditions. Regression analysis and variance analysis of the models demonstrated that each multinomial correctly represented the relationships between the responses and the variables. The urea‐to‐SOEE ratio was much more significant than crystallization temperature in UC, and the quadratic term of rotation speed of swept‐surface scrapers was the most significant variable in MD. Optimal UC conditions were 1.9:1 urea‐to‐SOEE ratio and ?1 °C crystallization temperature at which the purity and total recovery of EPA and DHA were 65.6 % and 46.8 %, respectively. The best conditions predicted for MD were 75 °C distillation temperature, 54.8 °C preheat temperature, 4.5 °C condensation temperature, and 307 rpm rotation speed at which the purity of EPA and DHA was 83.5 %. The predicted values were verified to be reasonably close to the experimentally observed values.     

Process conditions for separating fatty acid esters by supercritical CO2

The solubilities of ethyl palmitate, ethyl oleate, ethyl eicosapentaenoate (EPA) and ethyl docosahexaenoate (DHA) in supercritical carbon dioxide were determined by a continuous flow method. The solubilities of fatty acid ethyl esters increased with pressure and decreased as the temperature was increased. An empirical equation, similar to Chrastil's equation, was used to describe the relationship between solute solubility and the density of carbon dioxide. The empirical equation was further used to qualitatively estimate the separation efficiency of isolating EPA and DHA ethyl esters from fatty acid esters. The operating conditions yielding high solubility gave fast extraction rate but resulted in low separation efficiency. Experiments were conducted to separate ethyl EPA and ethyl DHA from a model mixture containing four fatty acid ethyl esters and from esterified squid visceral oil. The experimental data compared closely with the calculated values.     

Separation of fatty acid esters from cholesterol in esterified natural and synthetic mixtures by supercritical carbon dioxide

The solubility of cholesterol in supercritical carbon dioxide was determined by a continuous flow method. The solubility of cholesterol increased with increasing pressure and exhibited retrograde behavior. The Chrastil equation was used to describe the relationship between solubility and the density of carbon dioxide. A model mixture was made by adding cholesterol and fatty acid esters together. Squid visceral oil was esterified as the feed material. Both the model mixture and esterified squid visceral oil were extracted by supercritical carbon dioxide. The experimental results showed that cholesterol could be removed from a model mixture and from esterified squid visceral oil at low pressure (1500 psig) and high temperature (328.2°K). Under these conditions, cholesterol content in the extract was reduced from 2867 mg/100 g to 14.1 mg/100 g.     

Preparation of highly purified concentrates of eicosapentaenoic acid and docosahexaenoic acid

Because of the complexity of marine lipids, polyunsaturated fatty acid (PUFA) derivatives in highly purified form are not easily prepared by any single fractionation technique. The products are usually prepared as the ethyl esters by esterification of the body oil of fat fish species and subsequent physicochemical purification processes, including short-path distillation, urea fractionation, and preparative chromatography. Lipase-catalyzed transesterification has been shown to be an excellent alternative to traditional esterification and short-path distillation for concentrating the combined PUFA-content in fish oils. At room temperature in the presence of Pseudomonas sp. lipase and a stoichiometric amount of ethanol without any solvent, efficient transesterification of fish oil was obtained. At 52% conversion, a concentrate of 46% eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) was obtained in excellent recovery as a mixture of mono-, di-, and triacylglycerols. The latter can be easily separated from the saturated and monounsaturated ethyl esters and converted into ethyl esters either by conventional chemical means or enzymatically by immobilized Candida antarctica lipase. Urea-fractionation of such an intermediary product can give an EPA+DHA content of approximately 85%.     

Concentration and purification of stearidonic,eicosapentaenoic, and docosahexaenoic acids from cod liver oil and the marine microalgaIsochrysis galbana

n-3 Polyunsaturated fatty acids (n-3 PUFA) from the marine microalgaIsochrysis galbana were concentrated and purified by a two-step process—formation of urea inclusion compounds followed by preparative high-performance liquid chromatography. These methods had been developed previously with fatty acids from cod liver oil. By the urea inclusion compounds method, a mixture that contained 94% (w/w) stearidonic (SA), eicosapentaenoic (EPA), plus docosahexaenoic (DHA) acids (4:1 urea/fatty acid ratio and 4°C crystallization final temperature) was obtained from cod liver oil fatty acids. Further purification of SA, EPA, and DHA was achieved with reverse-phase C₁₈ columns. These isolations were scaled up to a semi-preparative column. A PUFA concentrate was isolated fromI. galbana with methanol/water (80:20, w/w) or ethanol/water (70:30, w/w). With methanol/water, a 96% EPA fraction with 100% yield was obtained, as well as a 94% pure DHA fraction with a 94% yield. With ethanol/water as the mobile phase, EPA and DHA fractions obtained were 92% pure with yields of 84 and 88%, respectively.     

Facile purification of tocopherols from soybean oil deodorizer distillate in high yield using lipase

Tocopherols have been purified from deodorizer distillate produced in the final deodorization step of vegetable oil refining by a process including molecular distillation. Deodorizer distillate contains mainly tocopherols, sterols, and free fatty acids (FFA); the presence of sterols hinders tocopherol purification in good yield. We found that Candida rugosa lipase recognized sterols as substrates but not tocopherols, and that esterification of sterols with FFA could be effected with negligible influence of water content. Enzymatic esterification of sterols with FFA was thus used as a step in tocopherol purification. High boiling point substances including steryl esters were removed from soybean oil deodorizer distillate by distillation, and the resulting distillate (soybean oil deodorizer distillate tocopherol concentrate; SODDTC) was used as a starting material for tocopherol purification. Several factors affecting esterification of sterols were investigated, and the reaction conditions were determined as follows: A mixture of SODDTC and water (4∶1, w/w) was stirred at 35°C for 24 h with 200 U of Candida lipase per 1 g of the reaction mixture. Under these conditions, approximately 80% of sterols was esterified, but tocopherols were not esterified. After the reaction, tocopherols and FFA were recovered as a distillate by molecular distillation of the oil layer. To enhance further removal of the remaining sterols, the lipase-catalyzed reaction was repeated on the distillate under the same reaction conditions. As a result, more than 95% of the sterols was esterified in total. The resulting reaction mixture was fractionated to four distillates and one residue. The main distillate fraction contained 65 wt% tocopherols with low contents of FFA and sterols. In addition, the residue fraction contained high-purity steryl esters. Because the process presented in this study includes only organic solvent-free enzymatic reaction and molecular distillation, it is feasible as a new industrial purification method of tocopherols. This work was presented at the Biocatalysis symposium in April 2000, held at the 91st Annual Meeting and Expo of the American Oil Chemists Society, San Diego, CA.     

Separation of eicosapentaenoic acid and docosahexaenoic acid in fish oil by kinetic resolution using lipase

The objective of this study was to investigate the use of lipases as catalysts for separating eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in fish oil by kinetic resolution. Transesterification of various fish oil triglycerides with a stoichiometric amount of ethanol by immobilized Rhizomucor miehei lipase under anhydrous solvent-free conditions resulted in a good separation. When free fatty acids from the various fish oils were directly esterified with ethanol under similar conditions, greatly improved results were obtained. By this modification, complications related to regioselectivity of the lipase and nonhomogeneous distribution of EPA and DHA into the various positions of the triglycerides were avoided. As an example, when tuna oil comprising 6% EPA and 23% DHA was transesterified with ethanol, 65% conversion into ethyl esters was obtained after 24 h. The residual glyceride mixture contained 49% DHA and 6% EPA (8:1), with 90% DHA recovery into the glyceride mixture and 60% EPA recovery into the ethyl ester product. When the corresponding tuna oil free fatty acids were directly esterified with ethanol, 68% conversion was obtained after only 8h. The residual free fatty acids comprised 74% DHA and only 3% EPA (25:1). The recovery of both DHA into the residual free fatty acid fraction and EPA into the ethyl ester product remained very high, 83 and 87%, respectively.     

Eicosapentaenoic acid is primarily responsible for hypotriglyceridemic effect of fish oil in humans

The aim of this study was to determine whether eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA), or both, were responsible for the triglyceride (TG)-lowering effects of fish oil. EPA (91% pure) and DHA (83% pure), a fish oil concentrate (FOC; 41% EPA and 23% DHA) and an olive oil (OO) placebo (all ethyl esters) were tested. A total of 49 normolipidemic subjects participated. Each subject was given placebo for 2–3 wk and one of the n-3 supplements for 3 wk in randomized, blinded trials. The target n-3 fatty acid (FA) intake was 3 g/day in all studies. Blood samples were drawn twice at the end of each supplementation phase and analyzed for lipids, lipoproteins, and phospholipid FA composition. In all groups, the phospholipid FA composition changed to reflect the n-3 FA given. On DHA supplementation, EPA levels increased to a small but significant extent, suggesting that some retroconversion may have occurred. EPA supplementation did not raise DHA levels, however, FOC and EPA produced significant decreases in both TG and very low density lipoprotein (VLDL) cholesterol (C) levels (P<0.01) and increases in low density lipoprotein (LDL) cholesterol levels (P<0.05). DHA supplementation did not affect cholesterol, triglyceride, VLDL, LDL, or high density lipoprotein (HDL) levels, but it did cause a significant increase in the HDL₂/HDL₃ cholesterol ratio. We conclude that EPA appears to be primarily responsible for TG-lowering (and LDL-C raising) effects of fish oil.     

Strategies for the enzymatic enrichment of PUFA from fish oil

PUFA from oil extracted from Nile perch viscera were enriched by selective enzymatic esterification of the free fatty acids (FFA) or by hydrolysis of ethyl esters of the fatty acids from the oil (FA‐EE). Quantitative analysis was performed using RP‐HPLC coupled to an evaporative light scattering detector (RP‐HPLC‐ELSD). The lipase from Thermomyces lanuginosus discriminated against docosahexaenoic acid (DHA) most, resulting in the highest DHA/DHA‐EE enrichment while lipase from Pseudomonas cepacia discriminated against eicosapentaenoic acid (EPA) most, resulting in the highest EPA/EPA‐EE enrichment. The lipases discriminated between DHA and EPA with a higher selectivity when present as ethyl esters (EE) than when in FFA form. Thus when DHA/EPA were enriched to the same level during esterification and hydrolysis reactions, the DHA‐EE/EPA‐EE recoveries were higher than those of DHA/EPA‐FFA. In reactions catalysed by lipase from T. lanuginosus, at 26 mol% DHA/DHA‐EE, DHA recovery was 76% while that of DHA‐EE was 84%. In reactions catalysed by lipase from P. cepacia, at 11 mol% EPA/EPA‐EE, EPA recovery was 79% while that of EPA‐EE was 92%. Both esterification of FFA and hydrolysis of FA‐EE were more effective for enriching PUFA compared to hydrolysis of the natural oil and are thus attractive process alternatives for the production of products highly enriched in DHA and/or EPA. When there is only one fatty acid residue in each substrate molecule, the full fatty acid selectivity of the lipase can be expressed, which is not the case with triglycerides as substrates.     

Production of n-3 polyunsaturated fatty acid-enriched fish oil by lipase-catalyzed acidolysis without solvent

Fish oil rich in n-3 polyunsaturated fatty acid (n-3 PUFA) was prepared by nonsolvent enzymic acidolysis. n-3 PUFA-enriched fish oil contained 25% eicosapentaenoic acid (EPA) and 40% docosahexaenoic acid (DHA). In acidolysis of cod liver oil, EPA content of the original fish oil was reduced at 5 h, but DHA content of the fish oil increased. It was assumed that EPA in the fish oil was replaced by DHA to reach a new chemical equilibrium. Two-stage acidolysis, which was carried out under CO₂ replacement early (about 3 h) and also in vacuum at 5–24 h, was effective for reduction in the content of diacylglycerol, which was formed by reverse reaction, hydrolysis. This method has industrial significance because PUFA-enriched triacylglycerol is easily separated from the reaction mixture by molecular distillation. Bioreactors for fats and their derivatives, Part XIV.     

Fractionation of menhaden oil ethyl esters using supercritical fluid CO2

Supercritical fluid CO₂ was used to fractionate menhaden oil fatty acid ethyl esters to obtain concentrates of the esters of allcis-5,8,11,14,17-eicosapentaenoic acid (EPA) and allcis-4,7,10,13,16,19-docosahexaenoic acid (DHA). Separation of the ethyl esters was found to occur primarily by carbon number, thus limiting the degree to which the ethyl esters of EPA and DHA could be concentrated. Urea fractionation of whole esters in order to remove saturates, monoenes and dienes prior to fractionation with supercritical fluid CO₂ resulted in concentrates of EPA and DHA in purities exceeding 90%. Several criteria are given for the selection of crude oils in order to maximize both purity and yield of concentrates.     

TMAH-Catalyzed Transesterification of EPA and DHA in Encapsulated Fish Oil Products

Tetramethylammonium hydroxide (TMAH)-catalyzed transesterification was developed as a rapid and reliable method using gas chromatography (GC) to determine the total fatty acid profile and to quantify the ethyl esters of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in 20 brands of encapsulated fish oil products. The AOAC method with boron trifluoride (BF₃) as a catalyst was used as a reference. After the respective transesterifications of BF₃ and TMAH, seven brands of encapsulated fish oil showed a single peak of EPA or DHA in the chromatograms, while 13 brands showed a single peak in the chromatograms after BF₃ esterification, but doublet peaks of EPA or DHA after TMAH esterification. By comparing with the GC/MS NIST library and authentic standard fatty acids of ethyl esters, the two pairs of doublet peaks were confirmed the ethyl and methyl esters of EPA and DHA, while the sum of the peak areas of the doublet represented the content of EPA or DHA. The reaction time course concluded that optimal TMAH transesterification was obtained at 25 °C for 10 min and using GC columns of low to medium polarity including Rtx-wax and Rtx-2330 were able to differentiate and quantify the ethyl- and methyl-esterified EPA and DHA, while RT-2560 column with higher polarity than the two other columns was unable to resolve the ethyl ester from the methyl ester of EPA or DHA. An EPA/DHA ratio of ≥1.10 may serve as an indicator of fish oil fortified with the ethyl ester of EPA.     

The preparation of concentrates of eicosapentaenoic acid and docosahexaenoic acid by lipase-catalyzed transesterification of fish oil with ethanol

The objective of this study was to investigate the use of lipases as catalysts for producing concentrates of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil as an alternative to conventional chemical procedures. Transesterification of fish oil with ethanol was conducted under anhydrous solvent-free conditions with a stoichiometric amount of ethanol. Among the 17 lipases tested, the results showed that Pseudomonas lipases had the highest activity toward the saturated and monounsaturated fatty acids in the fish oil, much lower activity toward EPA and DHA and, at the same time, good tolerance toward the anhydrous alcoholic conditions. With 10 wt% of lipase, based on weight of the fish oil triacylglycerol substrate (15% EPA and 9% DHA initial content), a 50% conversion into ethyl esters was obtained in 24 h at 20°C, in which time the bulk of the saturated and monounsaturated fatty acids reacted, leaving the long-chain n-3 polyunsaturated fatty acids unreacted in the residual mixture as mono-, di-, and triacylglycerols. This mixture comprised approximately 50% EPA+DHA. Total recovery of DHA and EPA was high, over 80% for DHA and more than 90% for EPA. The observed fatty acid selectivity, favoring DHA as a substrate, was most unusual because most lipases favor EPA.     

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.     

Purification of steryl esters from soybean oil deodorizer distillate

Soybean oil deodorizer distillate (SODD) contains steryl esters in addition to tocopherols and sterols. Tocopherols and sterols have been industrially purified from SODD but no purification process for steryl esters has been developed. SODD was efficiently separated to low b.p. substances (including tocopherols and sterols) and high b.p. substances (including 11.2 wt% DAG, 32.1 wt% TAG, and 45.4 wt% steryl esters) by molecular distillation. The high b.p. fraction is referred to as soybean oil deodorizer distillate steryl ester concentrate (SODDSEC). We attempted to purify steryl esters after a lipase-catalyzed hydrolysis of acylglycerols in SODDSEC. Screening of industrially available lipases indicated that Candida rugosa lipase was most effective. Based on the study of several factors affecting hydrolysis, the reaction conditions were determined as follows: ratio of SODDSEC/water, 1∶1 (w/w); lipase amount, 15 U/g reaction mixture; temperature, 30°C. When SODDSEC was agitated for 24 h under these conditions, acylglycerols were almost completely hydrolyzed and the content of steryl esters did not change. However, study with a mixture of steryl oleate/trilinolein (1∶1, w/w) indicated that about 20% of constituent FA in steryl esters were exchanged with constituent FA in acylglycerols. Steryl esters in the oil layer obtained by the SODDSEC treatment with lipase were successfully purified by molecular distillation (purity, 97.3%; recovery, 87.7%).     

Isolation of tocopherol and sterol concentrate from sunflower oil deodorizer distillate

The isolation of tocopherols and sterols together as a concentrate from sunflower oil deodorizer distillate was investigated. The sunflower oil deodorizer distillate was composed of 24.9% unsaponifiable matter with 4.8% tocopherols and 9.7% sterols, 28.8% free fatty acid (FFA) and 46.3% neutral glycerides. The isolation technology included process steps such as biohydrolysis, bioesterification and fractional distillation. The neutral glycerides of the deodorizer distillates were hydrolyzed byCandida cylindracea lipase. The total fatty acids (initial FFA plus FFA from neutral glycerides) were converted into butyl esters withMucor miehei lipase. The esterified product was then fractionally distilled in a Claisen-vigreux flask. The first fraction, which was collected at 180–230°C at 1.00 mm of Hg for 45 min, contained mainly butyl esters, hydrocarbons, oxidized products and some amount of free fatty acids. The fraction collected at 230–260°C at 1.00 mm Hg for 15 min was rich in tocopherols (about 30%) and sterols (about 36%). The overall recovery of tocopherols and sterols after hydrolysis, esterification and distillation were around 70% and 42%, respectively, of the original content in sunflower oil deodorizer distillate.     

Combined Urea Complexation and Argentated Silica Gel Column Chromatography for Concentration and Separation of PUFAs from Tuna Oil: Based on Improved DPA Level

Eicosapentaenoic acid (EPA, 20:5n‐3), docosapentaenoic acid (DPA) isomers (22:5n‐6 and 22:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3) derived from tuna oil were concentrated by three stages of urea fractionation at various crystallization temperatures and different fatty acid/urea ratios. Thereafter, polyunsaturated fatty acids concentrate containing comparatively enriched DPA levels was purified by argentated silica gel column chromatography. A product containing 22.2 ± 0.6 % EPA, 4.6 ± 0.0 % DPAn‐6, 5.9 ± 0.1 % DPAn‐3 and 42.3 ± 1.2 % DHA was obtained at 1:1.6 fatty acid/urea ratio (w/w) by crystallization at ?8 °C for 16 h, ?20 °C for 8 h, and ?8 °C for 16 h. A DPA isomer concentrate containing 26.1 ± 0.5 % DPAn‐6 and 22.3 ± 0.4 % DPAn‐3 was achieved by argentated silica gel chromatography in the 6 % acetone/n‐hexane solvent fraction (v/v), and the recovery of both fatty acids was 66.1 ± 3.2 and 70.7 ± 2.2 %, respectively. Furthermore, 91.9 ± 2.5 % EPA and 99.5 ± 2.1 % DHA with recoveries of 47.8 ± 2.0 and 56.7 ± 3.3 %, respectively, were obtained in various fractions.     

Wax Ester Rich Oil From The Marine Crustacean,Calanus finmarchicus,is a Bioavailable Source of EPA and DHA for Human Consumption

Oil from the marine copepod, Calanus finmarchicus, which contains >86 % of fatty acids present as wax esters, is a novel source of n‐3 fatty acids for human consumption. In a randomized, two‐period crossover study, 18 healthy adults consumed 8 capsules providing 4 g of Calanus^® Oil supplying a total of 260 mg EPA and 156 mg DHA primarily as wax esters, or 1 capsule of Lovaza^® providing 465 mg EPA and 375 mg DHA as ethyl esters, each with an EPA‐ and DHA‐free breakfast. Plasma EPA and DHA were measured over a 72 h period (t = 1, 2, 4, 6, 8, 10, 12, 24, 48, and 72 h). The positive incremental area under the curve over the 72 h test period (iAUC_{0‐72 h}) for both EPA and DHA was significantly different from zero (p < 0.0001) in both test conditions, with similar findings for the iAUC_{0–24 h} and iAUC_{0–48 h}, indicating the fatty acids were absorbed. There was no difference in the plasma iAUC_{0–72 h} for EPA + DHA, or DHA individually, in response to Calanus Oil vs the ethyl ester condition; however, the iAUC_{0–48 h} and iAUC_{0–72 h} for plasma EPA in response to Calanus Oil were both significantly increased relative to the ethyl ester condition (iAUC_{0–48 h}: 381 ± 31 vs 259 ± 39 μg*h/mL, p = 0.026; iAUC_{0‐72 h}: 514 ± 47 vs 313 ± 49 μg*h/mL, p = 0.009). These data demonstrate a novel wax ester rich marine oil is a suitable alternative source of EPA and DHA for human consumption.