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.     

Marked enrichment of the alkenylacyl subclass of plasma ethanolamine glycerophospholipid with eicosapentaenoic acid in human subjects consuming a fish oil concentrate

Alteration in human platelet fatty acid levels with the consumption of fish oils containing eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) have been well documented, but changes in the fatty acid composition of plasma plasmalogenic phospholipid under similar circumstances have not been delineated. In the present study, subjects consumed the fish oil concentrate (MaxEPA) for 6 wk followed immediately by a 6-wk recovery period with no fish oil ingestion. Plasma total choline glycerophospholipid (GPC) and ethanolamine glycerophospholipid (GPE) subclasses isolated from blood samples obtained at 0, 3, 6, 9 and 12 wk of the experimental period were analyzed for fatty acid composition via thin-layer and gas-liquid chromatographic techniques. Consumption of fish oil for 3 or 6 wk significantly elevated the content of n-3 fatty acids while concomitantly decreasing n-6 fatty acid levels in plasma total GPC and in diacyl and alkenylacyl (plasmalogen) GPE. Alkenylacyl GPE exhibited the greatest alteration of both n-3 and n-6 fatty acid levels. Following 6 wk of supplementation with fish oil, EPA rose by 24.6 mol% in alkenylacyl GPE compared to increases of 6.7 and 7.1 mol% in diacyl GPE and total GPC, respectively. The increase in EPA (from 5.0 to 29.6 mol%) in plasma alkenylacyl GPE represents amongst the highest enrichment of EPA in any lipid yet reported in human subjects. DHA also rose by 8.0, 4.8, and 3.1 mol% in alkenylacyl GPE, diacyl GPE, and total GPC, respectively. Alkenylacyl GPE exhibited the greatest mol% decline (by 18.7 mol%) in arachidonic acid (AA, 20:4n-6) following 6 wk of fish oil supplementation. The corresponding decreases of AA in diacyl GPE and total GPC were 8.7 and 1.8 mol%, respectively. Following the 6 wk recovery period, n-3 and n-6 fatty acid levels had returned to pre-supplementation values. The marked enrichment of alkenylacyl GPE in n-3 fatty acids, especially EPA, may be of significance with respect to a unique role for this plasma phospholipid subclass in attenuating certain lipoprotein-mediated cardiovascular effects as observed with fish/fish oil consumption.     

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.     

One-Step Concentration of Highly Unsaturated Fatty Acids from Tuna Oil by Low-Temperature Crystallization

Highly unsaturated fatty acids (HUFA), including eicosapentaenoic acid (EPA, 20:5n‐3), docosapentaenoic acid (DPA, 22:5n‐3 and 22:5n‐6) and docosahexaenoic acid (DHA, 22:6n‐3), play an important role in human health and nutrition. In this study, concentration of HUFA in free fatty acids (FFA) form by low‐temperature crystallization was investigated. For this purpose, tuna oil (7.1% EPA, 26.8% DHA) was first converted into corresponding FFA. Subsequently, crystallization conditions of various solvent types, the ratio of FFA to acetonitrile, operation temperature and crystallization time were optimized at a small scale of 2 g tuna oil fatty acids. Taking purity and yield into account, the optimum conditions were a 1:10 ratio of FFA to acetonitrile (w/v), ?60 °C, and 1 h. The optimal conditions resulted in concentrations of EPA, DHA and HUFA of 15.1, 58.4 and 79.6%, respectively, with corresponding yields of 61.5, 61.8 and 60.7%, respectively. Crystallization was carried out under the optimal conditions at a large scale of 200 g tuna oil FFA, and a similar concentration result was achieved. After evaporating away the solvent, the residual amount of acetonitrile met the US Pharmacopoeia requirement of <410 ppm. The process for enrichment of HUFA is readily scalable, effective and time‐saving.     

Kontinuierliches Verfahren zur Anreicherung von mehrfach ungesättigten Fettsäuren

Continuous Process for the Concentration of Polyunsaturated Fatty Acids Polyunsaturated fatty acids of the n-6 and n-3 series are of great nutritional interest. For special studies and applications, these acids ae required in high concentrations. Several acids of these series are, however, only available in concentrations of up to 25% in vegetable or animal oils. In an evaluation of different fractionation techniques with blackcurrant seed oil as example, reasonable results were obtained with urea fractionation in methanol. Applying this method, a specific separation of α- and γ-linolenic acid could be achieved, whereas stearidonic acid (C18 :4,n-3) had to separated from γ-linolenic acid (c18 :3,n-6) by subsequent preparative HPLC. During scaling up of this batch process to the ton scale, difficulties became apparent, requiring an increased number of fractionation steps, probably deu to insufficient heat exchange and to incomplete formation of occlusion crystals. These inconveniences were eliminated by developing a continuous process using heat exchangers with a scraped surface as reactors for urea occlusion formation. This continuous technique was also applied to other oils, as e. g. fish oil, borage oil and evening primrose oil.     

Effects of process parameters on EPA and DHA concentrate production from Atlantic salmon by-product oil: Optimization and characterization

Supercritical carbon dioxide (SC-CO₂) extracted Atlantic salmon frame bone oil (SFBO) was used for Eicosapentaenoic acid and Docosahexaenoic acid (EPA-DHA) concentrate production by urea complexation. Urea/fatty acids (2.5 to 4.0 w/w), crystallization temperature (?24 to ?8 °C) and crystallization time (8 to 24 h) were studied by Box-Behnken Design (BBD) to maximize EPA-DHA content. Highest EPA-DHA content was 60.63% at urea/fatty acids 4.0 w/w, crystallization temperature ?15.67 °C and crystallization time 8 h. EPA-DHA concentrate showed improvement of EPA-DHA from 6.39% in SFBO to 62.34%, increase of astaxanthin content from 21.33 μg/g in SFBO to 44.69 μg/g in EPA-DHA concentrate, no residual urea and reduction of many off-flavor compounds. The EPA-DHA yield showed an inverse relation with the urea/fatty acids, whereas its concentration increased proportionally with urea/fatty acids. Therefore, EPA-DHA concentrate produced from SFBO by urea complexation may be an efficient technique to provide ω-3 polyunsaturated fatty acids to the consumers.     

Fettsäuren-Zusammensetzung von Fischölkapseln

Fatty Acid Composition of Fish Oil Capsules The proven nutritional essentially of ω-fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) as well as their positive effect w.r.t. prevention and therapy of cardiovascular disease, have increased the significance of fish consumption as well as the intake of fish oil capsules from both marketing and medical points of view. Consequently, we analysed 26 fish oil preparation via GC, w.r.t. fatty acid composition as well as EPA- and DHA-content respectively. The aim was to identify the origin and provenance of the respective fish oils and thier possible blending will other edible fats and oils. The results of our investigations show that 23 samples could be considered as being composed of fish oils or fish oil concentrates. The remaining 3 were identified as being fish oil fatty acid ethylesters. An admixture with other edible oils could not be detected. Most of the fish oil capsules(= 20 products), as well as those specified as being salmon oil capsules, contained about 30% ω-3-fatty acids in a ratio between EPA:DHA of about 3:2. This ratio corresponds with those of a fish oil concentrate from Japan. Higher concentrations of ω-fatty acids of about 50% were found in the three capsules containing fish oil fatty acid ethylesters. The EPA:DHA ratio amounted in this case to about 3:2.     

Enrichment of Omega-3 Fatty Acids of Refined Hoki Oil

Enrichment of the omega-3 (n-3) fatty acids of refined hoki oil (RHO) intact triglycerides (TG) and via free fatty acids (FFA), was carried out in the present study using established methods of dry fractionation (DF), low temperature solvent crystallization (LTSC) and urea complexation (UC) and positional distribution of fatty acids in the intact TG was determined by Nuclear Magnetic Resonance (NMR) analysis. Results showed that n-3 fatty acids were enriched in liquid fractions of all methods except DF, where the highest concentration was obtained via the UC method (83.00 %). The FFA form of the oil produced a higher concentration (40.81 %) of n-3 fatty acids via the LTSC method compared to the TG form (31.50 %). The percentages of the total saturated fatty acid (SFA) in the liquid fractions in all methods were lower, ranging from 1.60 % (UC) to 21.44 % (DF) compared to the RHO parent oil (24.05 %). The percentages of total monounsaturated fatty acids (MUFA) in the liquid fractions were similar to the solid fractions except for the UC method where total MUFA was six times higher in the solid fraction. In LTSC-FFA and UC methods, the enrichment factor for EPA was lower, ranging from 1.61 (LTSC-FFA) to 2.83 (UC), than DHA which ranged from 1.64 (LTSC-FFA) to 3.88 (UC). EPA was preferentially located at the sn-1,3 position and DHA was significantly located at the sn-2 position which is the favoured location for intestinal digestion.     

Eicosapentaenoic acid (20∶5n-3) from the marine microalgaPhaeodactylum tricornutum

Eicosapentaenoic acid (EPA, 20∶5n-3) was obtained from the marine microalgaePhaeodactylum tricornutum by a three-step process: fatty acid extraction by direct saponification of biomass, polyunsaturated fatty acid (PUFA) concentration by formation of urea inclusion compounds, and EPA isolation by semipreparative high-performance liquid chromatography (HPLC). Alternatively, EPA was obtained by a similar two-step process without the PUFA concentration step by the urea method. Direct saponification of biomass was carried out with two solvents that contained KOH for lipid saponification. An increase in yield was obtained because the problems associated with emulsion formation were avoided by separating the biomass from the soap solution before adding hexane for extraction of insaponifiables. The most efficient solvent, ethanol (96%) at 60°C for 1 h, extracted 98.3% of EPA. PUFA were concentrated by the urea method with a urea/fatty acid ratio of 4∶1 at a crystallization temperature of 28°C and by using methanol and ethanol as urea solvents. An EPA concentration ratio of 1.73 (55.2∶31.9) and a recover yield of 78.6% were obtained with methanol as the urea solvent. This PUFA concentrate was used to obtain 93.4% pure EPA by semipreparative HPLC with a reverse-phase, C₁₈, 10 mm i.d.×25-cm column and methanol/water (1% acetic acid), 80∶20 w/w, as the mobile phase. Eighty-five percent of EPA loaded was recovered, and 65.7% of EPA present inP. tricornutum biomass was recovered in highly pure form by this three-step downstream process. Alternatively, 93.6% pure EPA was isolated from the fatty acid extract (without the PUFA concentration step) with 100% EPA recovery yield. This two-step process increases the overall EPA yield to 98.3%, but it is only possible to obtain 20% as much EPA as that obtained by three-step downstream processing.     

Lipase-assisted concentration of n-3 polyunsaturated fatty acids in acylglycerols from marine oils

Preparation of n-3 polyunsaturated fatty acid (PUFA) concentrates from seal blubber oil (SBO) and menhaden oil (MHO) in the form of acylglycerols was carried out by hydrolysis with a number of commercial microbial lipases. The lipases tested were Aspergillus niger, Candida cylindracea (CC), Chromobacterium viscosum, Geotrichum candidum, Mucor miehei, Pseudomonas sp., Rhizopus oryzae, and Rhizopus niveus. After lipase-assisted hydrolysis of oils, free fatty acids were removed, and fatty acid composition of the mixture containing mono-, di-, and triacylglycerols was determined. All lipases were effective in increasing the n-3 PUFA content of the remaining acylglycerols of both SBO and MHO. The highest concentration of n-3 PUFA was provided by CC lipase; 43.5% in SBO [9.75% eicosapentaenoic acid (EPA), 8.61% docosapentaenoic acid (DPA), and 24.0% docosahexaenoic acid (DHA)] and 44.1% in MHO (18.5% EPA, 3.62% DPA, and 17.3% DHA) after 40 h of hydrolysis. Thus, CC lipase appears to be most suitable for preparation of n-3 PUFA in the acylglycerol form from marine oils.     

Matrix Extension Validation of AOCS Ce 2c-11 for Omega-3 Polyunsaturated Fatty Acids in Conventional Foods and Dietary Supplements Containing Added Marine Oil

Marine oils are commonly added to conventional foods and dietary supplements to enhance their contents of omega-3 polyunsaturated fatty acids (PUFA), including eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3), which have been associated with numerous potential health benefits. This study compared American Oil Chemists’ Society (AOCS) Official Methods Ce 2b-11 and Ce 2c-11 for determining EPA and DHA in foods and dietary supplements and found that AOCS Ce 2c-11 produces significantly higher analyzed values, which could be attributed to a more comprehensive breakdown of the sample matrix and derivatization of fatty acids. Our subsequent food matrix extension validation of AOCS Ce 2c-11 demonstrated that the method produces true, accurate, sensitive, and precise determinations of EPA, DHA, and total omega-3 PUFA in foods and dietary supplements containing added marine oil, including those formulated with emulsified and microencapsulated oils. The method detection limits for EPA and DHA were 0.012 ± 0.002 and 0.011 ± 0.003 mg g⁻¹, respectively (means ± SD). The analyzed contents of EPA (1.26–386 mg serving⁻¹), DHA (1.37–563 mg serving⁻¹), and total omega-3 PUFA (2.69–1270 mg serving⁻¹) were reported for 27 conventional food and dietary supplement products. Eighteen products declared contents of DHA, EPA + DHA, or total omega-3 PUFA on product labels, and the analyzed contents of those fatty acids varied from 95 to 162% of label declarations for all but two of the products.     

Antioxidant Effect of Nanoemulsions Based on Citrus Peel Essential Oils: Prevention of Lipid Oxidation in Trout

The antioxidant effects of oil‐in‐water nanoemulsion based on edible citrus peel essential oils on the fatty acid composition of rainbow trout fillets stored at 4 ± 2 °C are investigated. Fish fillets are treated with nanoemulsion and stored for 16 days. Lipid samples are converted into fatty acid methyl esters which are then detected by gas chromatagrophy (GC). The results show that palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), vaccenic acid (C18:1?‐7), oleic acid (C18:1?9), eicosenoic acid (C20:1?9), linoleic acid (C18:2?6), linolenic acid (C18:3?3), eicosapentaenoic acid (EPA) (C20:5?3), and docosahexaenoic acid (DHA) (C22:6?3) are the most important fatty acids in fish meat. While polyene index and hypocholesterolemic:hypercholesterolaemic fatty acid ratios decrease in trout fillets during cold storage, thrombogenicity index and atherogenicity index generally increase (especially in control and Tween 80 groups). The concentrations of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) are higher in the treatment groups and the saturated fatty acids (SFAs) are lower in all groups compared to those of the control group. Application of nanoemulsion based on citrus essential oils prevents oxidation of PUFA especially EPA and DHA, thus has potential as a preservative for fish oil. Practical Applications: In recent years, nanotechnological applications have been increasingly applied to the protection of food. Similarly, natural essential oils are used to increase the shelf life of foods. This study demonstrates the combined effect of a new method of nanoemulsions and essential oils on the safety of foods.     

Malondialdehyde excretion by subjects consuming cod liver oil vs a concentrate of n-3 fatty acids

Urinary malondialdehyde (MDA), an indicator of lipid peroxidation in the diat and in the tissues, was determined in human adults consuming a supplement of n-3 fatty acids derived from a pharmaceutical grade of cod liver oil (CLO) without added antioxidants vs a concentrate of n-3 acids containing dodecyl gallate and vitamin E. MDA excretion increased immediately in the subjects consuming CLO but remained unchanged in those ingesting the concentrate for 50 days. The increase in the subjects taking CLO was attributable to MDA in the oil. The results indicate that consuming unstabilized fish oils as a source of n-3 fatty acids may entail exposure to potentially toxic products of lipid peroxidation.     

Enrichment of Omega-3 PUFAs from Fur Seal Oil

The solvent crystallization and the urea complexation of the Uruguayan fur seal oil (Arctocephalus australis Zim.), in order to obtain enriched omega-3 PUFAs concentrates were studied. The fractionation at –6°C of fur seal oil or its fatty acids dissolved in ethanol or acetone, is not suitable to obtain a PUFAs concentrate. Ethanol as a solvent and a two steps process (the second step consists of the addition of urea to the obtained NUCF) is the most useful procedure to obtain a concentrate of high PUFAs content using urea complexation of the free fatty acids from marine oils. When the concentrate is obtained from fur seal fatty acids, the total PUFAs content is of 90% with an overall yield of 17%. The recovery efficiency of total PUFAs is 78%. This procedure is very simple and relatively cheap.     

Incorporation of n-3 fatty acids into plasma lipid fractions, and erythrocyte membranes and platelets during dietary supplementation with fish, fish oil, and docosahexaenoic acid-rich oil among healthy young men

The effects of n-3 fatty acid supplementation in the form of fresh fish, fish oil, and docosahexaenoic acid (DHA) oil on the fatty acid composition of plasma lipid fractions, and platelets and erythrocyte membranes of young healthy male students were examined. Altogether 59 subjects (aged 19–32 yr, body mass index 16.8–31.3 kg/m²) were randomized into the following diet groups: (i) control group; (ii) fish diet group eating fish meals five times per week [0.38±0.04 g eicosapentaenoic acid (EPA) and 0.67±0.09 g DHA per day]; (iii) DHA oil group taking algae-derived DHA oil capsules (1.68 g/d DHA oil group taking algae-derived DHA oil capsules (1.68 g/d DHA in triglyceride form); and (iv) fish oil group (1.33 g EPA and 0.95 g DHA/d as free fatty acids) for 14 wk. The fatty acid composition of plasma lipids, platelets, and erythrocyte membranes was analyzed by gas chromatography. The subjects kept 4-d food records four times during the study to estimate the intake of nutrients. In the fish diet, in DHA oil, and in fish oil groups, the amounts of n-3 fatty acids increased and those of n-6 fatty acids decreased significantly in plasma lipid fractions and in platelets and erythrocyte membranes. A positive relationship was shown between the total n-3 polyunsaturated fatty acids (PUFA) and EPA and DHA intake and the increase in total n-3 PUFA and EPA and DHA in all lipid fractions analyzed. DHA was preferentially incorporated into phospholipid (PL) and triglyceride (TG) and there was very little uptake in cholesterol ester (CE), while EPA was preferentially incorporated into PL and CE. The proportion of EPA in plasma lipids and platelets and erythrocyte membranes increased also by DHA supplementation, and the proportion of linoleic acid increased in platelets and erythrocyte membranes in the DHA oil group as well. These results suggest retroconversion of DHA to EPA and that DHA also interferes with linoleic acid metabolism.     

The production of fish oils enriched in polyunsaturated fatty acid-containing triglycerides

A simple concentration technique was developed and used for the production of fish oils highly enriched with respect to the polyunsaturated triglycerides. The method involves the rapid solidification of fish oil droplets in liquid nitrogen followed by extraction with acetone at −60°C. The combined percentage ofcis-5,8,11,14,17-eicosapentaenoic acid (20:5) andcis-4,7,10,13,16,19-docosahexaenoic acid (22:6) after enrichment of a crude South African Anchovy (Engraulis capensis) oil was 57.4. A maximum percentage of 66.0 was attained for n-3 fatty acids after enrichment of a crude Chilean fish oil. A maximum yield of 26.0% was achieved for a crude sardine (Sardina pilchardus) oil. Triglycerides containing only saturated fatty acids or a combination of saturated and monoenoic acids were totally removed by the process, as assessed by silver-ion high-performance liquid chromatography of the triglyceride oils. This process permits the production of significant quantities of highly unsaturated triglycerides, which may be used in physiological and oxidative studies or for structural analysis of these triglycerides, many of which are present at extremely low concentrations in the original oils.     

Enrichment of n-3 containing ether phospholipids in plasma after 30 days of krill oil compared with fish oil supplementation

There are conflicting findings over the bioavailability of long-chain n-3 polyunsaturated fatty acids (n-3 PUFA) from krill oil (KO) compared with fish oil (FO) in short- and long-term studies. The aim of this study was to compare the effects of KO versus FO on the enrichment of molecular species of plasma phospholipids in young women following a 30-day consumption of the n-3 oils. Eleven healthy women aged 18–45 years consumed seven capsules of KO per day (containing a total of 1.27 g n-3 PUFA) or five capsules of FO per day (total of 1.44 g n-3 PUFA) for 30 days in a randomized crossover study, separated by at least a 30-day washout period. Fasting blood samples were collected at day zero (baseline), day 15 and day 30 and analyzed by HPLC-MS/MS for molecular species of phospholipids. Supplementation increased n-3 PUFA in main phospholipids classes in both groups. After 30 days of supplementation, 35 out of 70 molecular species containing eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and docosapentaenoic acid (DPAn-3) had a significantly greater concentration in KO group compared with the FO treated group. The majority (89%) of the differentiated molecular species were choline and ethanolamine ether-phospholipids. These data reveal that analysis of plasma phospholipids following 30 days of consumption of KO (a marine oil rich in phospholipids, including ether phospholipids) resulted in an enrichment of n-3 PUFA in molecular species of ether-phospholipids compared with FO (a triacylglycerol-rich marine oil).     

Effect of degree of unsaturation in dietary fatty acids on arachidonic acid mobilization by peritoneal macrophages

Cells from rats fed with a tripalmitin diet showed a depletion of phospholipid arachidonate and n-3 fatty acids such as eicosapentaenoic and docosahexaenoic acids (EPA and DHA). In rats fed fish oil diet, a significant reduction in archidonic acid (AA) content was observed whereas EPA and DHA were incorporated into membranes lipids. These changes in lipid composition of membranes did not affect cellular adherence, phagocytic capability, or [³H]AA incorporation. However, both tripalmitin and fish oil diets induced a decrease in [³H]AA mobilization stimulated by 4β-phorbol-12-myristate 13-acetate, A23187, or opsonized-zymosan in rat peritoneal macrophages. These results demonstrate that the antiinflammatory effects of essential fatty acids deficiency or n-3 enrichment diets may be associated with a decreased AA mobilization in resident rat peritoneal macrophages treated with proinflammatory agents.     

Preparation of eicosapentaenoic acid (EPA) concentrate fromPorphyridium cruentum

The polyunsaturated fatty acid eicosapentaenoic acid (EPA) has attracted increased attention due to its pharmaceutical properties. The main source is marine fish oil which contains approximately 15% EPA. However, pharmaceutical applications of EPA will probably require higher concentrations, perhaps as high as 90%. The red microalgaPorphyridium cruentum is a potential source, because its EPA content approaches 44.1% of the total fatty acids. Three methods were attempted for EPA concentration and arachidonic acid (AA) removal from the oil of this alga. Separation of the glycolipids, formation of a urea inclusion complex and reverse phase chromatography on C-18 Sep-Pak filters resulted in an EPA concentrate of 97% purity. Similar methods resulted in an AA concentrate of 80% purity.     

Fast Transmethylation of Serum Lipids Using Microwave Irradiation

Microwave irradiation as the energy source for one-step direct transesterification of fatty acids in human serum lipids was examined in a solvent system of methanol: hexane: acetyl chloride based on a Lepage & Roy assay. Innovative and explosion proof single-mode or multimode microwave accelerated reaction system was employed. Recoveries were calculated as the percentage of fatty acid concentrations measured by microwave assay to those by the reference method of the Lepage & Roy assay that utilized conductive heating at 100?°C for 60?min. Under conditions of 100?°C for 1?min in Single-mode (S4–100?×?1), or 125?°C for 5?min in Multimode (M5–125?×?5), the recoveries were 100–103?% for the total fatty acids and 96–106?% for each categorized fatty acid, including saturates, monounsaturates, n-6 PUFA, and n-3 PUFA. For individual PUFA, the mean recoveries were 102–105?% for 18:2n-6 and 18:3n-3; 99, 109, and 95?% for 20:4n-6, 20:5n-3, and 22:6n-3, respectively. Thus, fatty acid concentrations determined by microwave fatty acid assay were accurate to those results by the reference method, when the microwave conditions were optimal. In summary, the microwave irradiation could replace conductive heating in one-step direct transesterification, and reduce the duration from 60?min to 5?min or less. This methodology may be applied in both the absolute and relative quantification of serum total fatty acids.