Comparative Effects of Sandalwood Seed Oil on Fatty Acid Profiles and Inflammatory Factors in Rats

The aim of the present study was to investigate the effect of sandalwood seed oil on fatty acid (FA) profiles and inflammatory factors in rats. Fifty male Sprague–Dawley rats were randomly divided into five different dietary groups: 10 % soybean oil (SO), 10 % olive oil (OO), 10 % safflower oil (SFO), 10 % linseed oil (LSO) and 8 % sandalwood seed oil blended with 2 % SO (SWSO) for 8 weeks. The SWSO group had a higher total n-3 polyunsaturated fatty acids (PUFA) levels but lower n-6:n-3 PUFA ratios in both adipose tissue and liver than those in the SO, OO and SFO groups (p < 0.05). Although the SWSO group had a much lower 18:3n-3 level (4.51 %) in their dietary lipids than the LSO group (58.88 %), the levels of docosahexaenoic acid (DHA: 22:6n-3) in liver lipids and phospholipids of the SWSO group (7.52 and 11.77 %) were comparable to those of the LSO group (7.07 and 13.16 %). Ximenynic acid, a predominant acetylenic FA in sandalwood seed oil, was found to be highly incorporated into adipose tissue (13.73 %), but relatively lower in liver (0.51 %) in the SWSO group. The levels of prostaglandin F_2α, prostaglandin E₂, thromboxane B₂, leukotriene B₄, tumor necrosis factor-α and interleukin-1β in both liver and plasma were positively correlated with the n-6:n-3 ratios, suggesting that increased n-6 PUFA appear to increase the formation of pro-inflammatory cytokines, whereas n-3 PUFA exhibit anti-inflammatory activity. The present results suggest that sandalwood seed oil could increase tissue levels of n-3 PUFA, DHA and reduce the n-6:n-3 ratio, and may increase the anti-inflammatory activity in rats.     

Non-methylene Interrupted and Hydroxy Fatty Acids in Polar Lipids of the Alga Grateloupia turuturu Over the Four Seasons

Phospholipids (PL) and glycolipids (GL) FA in the edible Rhodophyta Grateloupia turuturu, from Brittany, France, were investigated over four seasons. The major lipid class was GL in all seasons (around 45 %). More than 80 FA occurred in polar lipids, with chains from C₁₂ to C₂₆, identified as methyl esters and N-acyl pyrrolidides by gas chromatography–mass spectrometry (GC–MS). PUFA occurred at up to 47.1 % (summer) in PL, and up to 43.6 % (summer) in GL. The major PUFA were 20:5n-3 (12.2 % in PL and 29.0 % in GL) and 20:4n-6 (25.6 % in PL and 10.4 % in GL). The unusual 18:3n-7 acid was identified in PL up to 2.2 %. Several minor unsaturated FA were identified in PL and are previously unreported in seaweeds, namely 14-tricosenoic, 15-tetracosenoic, 5,11-octadecadienoic and 5,9-nonadecadienoic. Also unprecedented in seaweeds, ten 2-hydroxy and three 3-hydroxy FA occurred mainly in PL, 13.9 % in spring with the 3-hydroxyhexadecanoic acid as the major one (8.1 % winter). Three n-9 monounsaturated 2-hydroxy FA occurred in PL. The 2-hydroxy-15-tetracosenoic acid was characterized as the dimethyl disulfide adduct of its methyl ester. The 2-hydroxy-16-pentacosenoic and 2-hydroxy-17-hexacosenoic acids were identified by comparison of mass spectra and GC mobilities with those of the 2-hydroxy-15-tetracosenoic acid, and of other homogeneous FA series. These rare n-9 monounsaturated 2-hydroxy FA are unprecedented in seaweeds.     

UVB Radiation Variably Affects n-3 Fatty Acids but Elevated Temperature Reduces n-3 Fatty Acids in Juvenile Atlantic Salmon (Salmo salar)

Temperature and ultraviolet B radiation (UVB 290–320?nm) are inextricably linked to global climate change. These two variables may act separately, additively, or synergistically on specific aspects of fish biochemistry. We raised Atlantic Salmon (Salmo salar) parr for 54?days in outdoor tanks held at 12 and 19?°C and, at each temperature, we exposed them to three spectral treatments differing in UV radiation intensity. We quantified individual fatty acid (FA) mass fractions in four tissues (dorsal muscle, dorsal and ventral skin, and ocular tissue) at each temperature?×?UV combination. FA composition of dorsal muscle and dorsal and ventral skin was not affected by UV exposure. Mass fractions of 16:0, 18:0, and saturated fatty acids (SFA) were greater in dorsal muscle of warm-reared fish whereas 18:3n-3, 20:2, 20:4n-6, 22:5n-3, 22:6n-3, n-3, n-6, polyunsaturated fatty acids (PUFA), and total FA were significantly higher in cold-reared fish. Mass fractions of most of the FA were greater in the dorsal and ventral skin of warm-reared fish. Cold-reared salmon exposed to enhanced UVB had higher ocular tissue mass fractions of 20:2, 20:4n-6, 22:6n-3, n-3, n-6, and PUFA compared to fish in which UV had been removed. These observations forecast a host of ensuing physiological and ecological responses of juvenile Atlantic Salmon to increasing temperatures and UVB levels in native streams and rivers where they mature before smolting and returning to the sea.     

Muscle Lipids and Fatty Acid Profiles of the Sea Catfish (Arius madagascariensis) in Madagascar Inland Waters

Muscle lipids and fatty acids (FA) of catfish Arius madagascariensis were determined in catfish caught in the Betsiboka River, Madagascar, during a 5-month sampling period. Total lipids from muscle were extracted and quantified. Fatty acids were identified by means of gas chromatography–mass spectrometry of FA methyl esters and FA pyrrolidides, leading to the identification of 42 FA. Lipid content was relatively high in our fish sample and ranged from 4.3 to 6.6% of wet muscle. Three FA dominated the FA composition: palmitic acid (C16:0, 22.9–32.6%), oleic acid (C18:1n-9, 11.3–13.4%) and stearic acid (C18:0, 10.8–12.0%). A number of polyunsaturated FA (PUFA) were present in appreciable amounts, including arachidonic acid (C20:4n-6, 4.7–7.6%), docosahexaenoic acid (C22:4n-6, 3.0–8.1%), eicosapentaenoic acid (C20:5n-3, 0.6–1.0%), n-3 docosapentaenoic acid (C22:5n-3, 1.1–1.6%), n-6 docosatetraenoic acid (C22:4n-6, 0.7–1.2%) and n-6 docosapentaenoic acid (22:5n-6, 0.9–1.8%). The sum of the n-6 PUFA and n-3 PUFA was 11.3–18.8 and 7.5–13.4%, respectively. These results indicate that A. madagascariensis, an abundant freshwater fish in Madagascar rivers, may be good source of dietary PUFA.     

Evaluation of the Stability of Fatty Acids in Erythrocytes from Human Umbilical Cord

The interest in the amount of polyunsaturated fatty acids (PUFA) in the umbilical cord blood (UCB) is increasing, but the stability of erythrocyte PUFA in these samples during storage and washing of the erythrocytes has not been directly evaluated. The purpose of this study was to analyze the effect of the lapse of time on the fatty acid (FA) content from UCB sample collection and maintained at 4 °C (0–12 h) until erythrocyte separation and washing. Palmitic acid (16:0), stearic acid (18:0), 18:1n-7/n-9, linoleic acid (18:2n-6), arachidonic acid (20:4n-6), 22:4n-6, eicosapentaenoic acid (20:5n-3), docosapentaenoic acid (22:5n-3), and docosahexaenoic acid (22:6n-3) together accounted for 87% of the FA profile in the umbilical vein erythrocytes. No difference was observed in the concentration of any of the FA studied, nor in the sum of saturated fatty acids (SFA), PUFA, or LC-PUFA in umbilical erythrocytes obtained at delivery and stored up to 12 h before the separation of erythrocytes. However, if a washing step was included in the processing of the erythrocytes, a decrease in the concentration of 16:0, 18:0, 18:3n-3, 20:4n-6, 22:4n-6, total SFA, PUFA, LC-PUFA, and n-6 LC-PUFA was evidenced, compared to unwashed erythrocytes. The FA concentration in umbilical cord erythrocytes did not change between samples stored from 0 to 12 h until erythrocyte separation. Erythrocyte washing before storage decreased the concentration of significant individual and total SFA, PUFA, and LC-PUFA. These results should be considered when planning the collection of UCB samples for the study of fatty acid concentration due to the nonscheduled timing of deliveries.     

Retroconversion of Docosapentaenoic Acid (n-6): an Alternative Pathway for Biosynthesis of Arachidonic Acid in Daphnia magna

The aim of this study was to assess metabolic pathways for arachidonic acid (20:4n-6) biosynthesis in Daphnia magna. Neonates of D. magna were maintained on [¹³C] enriched Scenedesmus obliquus and supplemented with liposomes that contained separate treatments of unlabeled docosapentaenoic acid (22:5n-6), 20:4n-6, linoleic acid (18:2n-6) or oleic acid (18:1n-9). Daphnia in the control treatment, without any supplementary fatty acids (FA) containing only trace amounts of 20:4n-6 (~0.3 % of all FA). As expected, the highest proportion of 20:4n-6 (~6.3 %) was detected in Daphnia that received liposomes supplemented with this FA. Higher availability of 18:2n-6 in the diet increased the proportion of 18:2n-6 in Daphnia, but the proportion of 20:4n-6 was not affected. Daphnia supplemented with 22:5n-6 contained ~3.5 % 20:4n-6 in the lipids and FA specific stable isotope analyses validated that the increase in the proportion of 20:4n-6 was due to retroconversion of unlabeled 22:5n-6. These results suggest that chain shortening of 22:5n-6 is a more efficient pathway to synthesize 20:4n-6 in D. magna than elongation and desaturation of 18:2n-6. These results may at least partially explain the discrepancies noticed between phytoplankton FA composition and the expected FA composition in freshwater cladocerans. Finally, retroconversion of dietary 22:5n-6 to 20:4n-6 indicates Daphnia efficiently retain long chain n-6 FA in lake food webs, which might be important for the nutritional ecology of fish.     

Peripheral Artery Disease Is Associated with a Deficiency of Erythrocyte Membrane n-3 Polyunsaturated Fatty Acids

Population-based data suggest that individuals who consume large dietary amounts of n-3 polyunsaturated fatty acids (PUFA) have lower odds of peripheral artery disease (PAD); however, clinical studies examining n-3 PUFA levels in patients with PAD are sparse. The objective of this study is to compare erythrocyte membrane fatty acid (FA) content between patients with PAD and controls. We conducted a cross-sectional study of 179 vascular surgery outpatients (controls, 34; PAD, 145). A blood sample was drawn and the erythrocyte FA content was assayed using capillary gas chromatography. We calculated the ratio of the n-3 PUFA eicosapentaenoic acid (EPA) to the n-6 PUFA arachidonic acid (ARA) as well as the omega-3 index (O3I), a measure of erythrocyte content of the n-3 PUFA, EPA, and docosahexaenoic acid (DHA), expressed as a percentage of total erythrocyte FA. Compared with controls, patients with PAD smoked more and were more likely to have hypertension and hyperlipidemia (p < 0.05). Patients with PAD had a lower mean O3I (5.0 ± 1.7% vs 6.0 ± 1.6%, p < 0.001) and EPA:ARA ratio (0.04 ± 0.02 vs 0.05 ± 0.05, p < 0.001), but greater mean total saturated fats (39.5 ± 2.5% vs 38.5 ± 2.6%, p = 0.01). After adjusting for several patient characteristics, comorbidities, and medications, an absolute decrease of 1% in the O3I was associated with 39% greater odds of PAD (odds ratio [OR] 1.39, 95% confidence interval [CI] 1.03–1.86, and p = 0.03). PAD was associated with a deficiency of erythrocyte n-3 PUFA, a lower EPA:ARA ratio, and greater mean total saturated fats. These alterations in FA content may be involved in the pathogenesis or development of poor outcomes in PAD.     

Fatty Acids in Liver,Muscle and Gonad of three Tropical Rays including Non-Methylene-Interrupted Dienoic Fatty Acids

Scientific investigation of lipids in Elasmobranchs has been conducted mainly on shark species. Because rays seem to be neglected, this study was performed to examine the complete fatty acid (FA) composition with a particular interest for long-chain polyunsaturated FA (PUFA) content in different tissues of three ray species including parts usually discarded. The total FA and PUFA profiles of total lipids were determined in muscle, liver, and gonad of Rhinobatos cemiculus, Rhinoptera marginata, and Dasyatis marmorata, the most often caught ray species from the East Tropical Atlantic Ocean. Fifty FA were characterized as methyl esters and N-acyl pyrrolidides by gas chromatography/mass spectrometry, showing significant levels of 20:5n-3 (EPA) (up to 5.3%) and 22:5n-3 (DPA) (up to 7.3%), high levels of 20:4n-6 arachidonic (ARA) (4.8–8.6% of total FA) and 22:6n-3 (DHA) (up to 20.0%). The results show that muscle, liver and gonad of rays can provide high amounts of essential PUFA, specially DHA, for direct human nutrition or the food processing industry. High proportions of DHA were particularly found in all samples of R. cemiculus (11.6–20.0%), and in muscle and liver of D. marmorata (11.1–16.1%). Regarding the high amounts of (n-3) PUFA, this study shows that these rays deserve a better up-grading, including the normally discarded parts, and describes the occurrence of unusual NMID FA in all tissues studied. Five non-methylene-interrupted dienoic fatty acids (NMID FA) (0–3.4%) were reported, including previously known isomers, namely 20:2 Δ7,13, 20:2 Δ7,15, 22:2 Δ7,13, 22:2 Δ7,15, and new 22:2 Δ6,14. These acids are quite unusual in fish and unprecedented in rays. The 22:2 Δ6,14 acid occurred in gonads of male specimens of R. cemiculus at 2.9%.     

Lipid Content and Fatty Acid Profile of Muscle,Brain and Eyes of Seven Freshwater Fish: a Comparative Study

The fatty acid and lipid content of the eyes, brain and muscle were determined for the following seven freshwater fish species in China: black carp, grass carp, silver carp, bighead carp, common carp, crucian carp, and Wuchang bream. Lipid contents of all seven freshwater fish were in the order of brain > eyes > muscle. Most of the freshwater fish demonstrated much lower n-3 PUFA than n-6 PUFA in all tissues with n-3/n-6 ratios varying between 0.07 and 0.40, while silver carp and bighead carp had much higher n-3 PUFA than n-6 PUFA. The percentage of PUFA in the muscle of all species are the highest compared to those of the eyes and brain, while no significant difference was observed in PUFA concentration between eyes and brain (p > 0.05). Multivariable analyses indicate that the fatty acid composition of the filter-feeder fish (silver carp and bighead carp) was distinct from the other four species.     

High Arachidonic Acid Levels in the Tissues of Herbivorous Fish Species (<Emphasis Type="Italic">Siganus fuscescens,Calotomus japonicus</Emphasis> and <Emphasis Type="Italic">Kyphosus bigibbus</Emphasis>)

The lipid and fatty acid compositions in the various organs (muscle, liver, other viscera) and stomach contents of three common herbivorous fish species in Japan, Siganus fuscescens, Calotomus japonicus and Kyphosus bigibbus, were examined to explore the stable 20:4n-6 (arachidonic acid, ARA) sources. Triacylglycerol (TAG), phosphatidylethanolamine (PtdEtn), and phosphatidylcholine (PtdCho) were the dominant lipid classes, while the major FA contents were 16:0, 18:1n-9, 16:1n-7, 14:0, 18:0, 18:1n-7, and some PUFA, including ARA, 20:5n-3 (eicosapentaenoic acid, EPA), 22:5n-3 (docosapentaenoic acid, DPA), and 22:6n-3 (docosahexaenoic acid, DHA). The amounts of these fatty acids were varied among species and their lipid classes. Phospholipids contained higher levels of PUFA than TAG. However, ARA in both phospholipids and TAG was markedly present in the muscle and viscera of all specimens, particularly in C. japonicus and K. bigibbus. Moreover, their ARA levels were higher than the levels of DHA and EPA. The observed high ARA level is unusual in marine fish and might be characteristic of herbivorous fish. Furthermore, ARA was the dominant PUFA in the stomach contents of the three species, suggesting that the high ARA level originated from their food sources. The above indicates that these three herbivorous fishes are ARA-rich marine foods and have potential utilization as stable ARA resources.     

Fatty Acid Profile of Sunshine Bass: I. Profile Change is Affected by Initial Composition and Differs Among Tissues

Fatty acid (FA) composition of fillet tissue can be tailored by transitioning fish from alternative lipid-based, low long-chain polyunsaturated fatty acid (LC-PUFA) grow-out feeds to high LC-PUFA "finishing" feeds. To address whether grow-out feed composition influences the responsiveness of fillet tissue to finishing, sunshine bass (SB, Morone chrysops x M. saxatilis) were reared to a submarketable size on grow-out feeds containing fish oil (FO) or a 50:50 blend of FO and coconut (CO), grapeseed (GO), linseed (LO), or poultry (PO) oil. For the final 8 weeks of the trial, fish were either maintained on assigned grow-out feeds or finished with the 100% FO feed. Production performance was unaffected by dietary lipid source, but fillet FA profile generally conformed to nutritional history. Regardless of grow-out regimen, finishing had a significant restorative effect on fillet FA composition; however, complete restoration of control levels of 20:5n-3, 22:6n-3, total LC-PUFA and n-3:n-6 FA ratio was achieved only among fish fed the CO-based grow-out feed. Saturated fatty acids (SFA) appear to be preferential catabolic substrates, whereas medium-chain and long-chain PUFA are selectively deposited in tissues. Provision of SFA in grow-out feeds appears to optimize selective FA metabolism and restoration of beneficial fillet FA profile during finishing.     

Stearidonic Acid Increases the Red Blood Cell and Heart Eicosapentaenoic Acid Content in Dogs

Plant sources of omega-3 fatty acids (FA) are needed that can materially raise tissue levels of long-chain omega-3 FA [i.e., eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 20:6n-3)]. Stearidonic acid (SDA; 18:4n-3) is the delta-6 desaturase product of alpha-linolenic acid (ALA; 18:3n-3), and when fed to humans, increases red blood cell (RBC) content of EPA to a greater extent than does ALA. This study was undertaken to determine the dose-dependence and time course of the increase in the EPA and DHA content of the heart and RBC in dogs. Adult male Beagles were fed 21, 64, or 193 mg/kg of SDA in in their food daily for up to 12 weeks. Positive and negative controls were given EPA (43 mg/kg) or high oleic acid sunflower oil, respectively. The baseline EPA content of RBC was 0.38 ± 0.03% which increased (P < 0.01) in a dose-dependent manner, with the high dose of SDA and EPA achieving levels of 1.33 ± 0.26 and 1.55. ± 0.28%, respectively. In the heart, the content of EPA rose from 0.06 ± 0.01 to 1.24 ± 0.22% in the EPA group and to 0.81 ± 0.32% in the high SDA group (both P < 0.01). In both tissues, DHA did not change. Compared to dietary EPA, SDA was 20–23% as efficient in raising tissue EPA levels. In conclusion, SDA supplementation increased the EPA content of RBC and heart and may have utility as a plant-based source of omega-3 FA.     

Essential Polyunsaturated Fatty Acids in Blood from Patients with and without Catheter-Proven Coronary Artery Disease

Coronary artery disease (CAD) is the leading cause of death worldwide. Statins reduce morbidity and mortality of CAD. Intake of n-3 polyunsaturated fatty acid (n-3 PUFAs), particularly eicosapentaenoic acid (EPA), is associated with reduced morbidity and mortality in patients with CAD. Previous data indicate that a higher conversion of precursor fatty acids (FAs) to arachidonic acid (AA) is associated with increased CAD prevalence. Our study explored the FA composition in blood to assess n-3 PUFA levels from patients with and without CAD. We analyzed blood samples from 273 patients undergoing cardiac catheterization. Patients were stratified according to clinically relevant CAD (n = 192) and those without (n = 81). FA analysis in full blood was performed by gas chromatography. Indicating increased formation of AA from precursors, the ratio of dihomo-gamma-linolenic acid (DGLA) to AA, the delta-5 desaturase index (D5D index) was higher in CAD patients. CAD patients had significantly lower levels of omega-6 polyunsaturated FAs (n-6 PUFA) and n-3 PUFA, particularly EPA, in the blood. Thus, our study supports a role of increased EPA levels for cardioprotection.     

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.     

Production of PUFA Concentrates from Poultry and Fish Processing Waste

In fish and poultry processing, viscera are generally considered as a waste product and often discarded. Chicken and hilsa fish (Hilsa ilisa) viscera were used for the production of polyunsaturated fatty acids (PUFA) linoleic (18:2n-6), eicosapentaenoic (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). Free fatty acids (FFA) were extracted by alkaline hydrolysis of chicken and fish viscera; yields were 5.2 and 5.9% (w/w) respectively. PUFA concentrates were obtained by a two step process—deduction of saturated fatty acids (FA) by low temperature crystallization in acetone followed by urea inclusion compound-based fractionation. Acetone treatment removed 90 and 96% of saturated FA in chicken and fish viscera respectively with FA to acetone ratio of 1:12 (w/v). Using an urea to FA ratio (w/w) of 4.0, chicken viscera produced a maximum of 84.1% of PUFA concentrates containing 82.1% of linoleic acid with a yield of 10% where as in the case of fish viscera the maximum PUFA concentrates were 91.3% containing 78.2% of EPA-DHA with the yield of 11%. Thus, the utilization of poultry and fish processing waste into the production of PUFA concentrates has been shown.     

Fatty Acid Contents in Fractions of Neutral Lipids and Phospholipids of Fillets of Tilapia Treated with Flaxseed Oil

The fatty acid composition in fractions of neutral lipids (NL) and phospholipids (PL) of the lipid content of muscle tissue of tilapia (Oreochromis niloticus) fed diets enriched with 7% of flaxseed oil was evaluated. Lipid profiles were evaluated every 15 days over 90 days. NL were predominant, with saturated and monounsaturated fatty acids being the major ones, and the PL fraction showed a great amount of polyunsaturated fatty acids (PUFA). The PUFA/SFA ratio values were lower in the NL fraction, ranging from 0.3 to 0.6, when compared to the PL fraction, which ranged from 1.0 to 1.1. The n-3 PUFA concentrations increased from the beginning of the treatment up to 90 days of treatment, while the n-6 PUFA concentrations decreased in the same period, thus improving the n-6/n-3 ratio of lipid content. The n-6/n-3 ratios in the NL class varied from 11.1 to 1.1 and in the PL class from 4.6 to 1.0, which contributed to an increase in the nutritional value of the fish meat. The PUFA/SFA ratio values increased during the treatment, whereas the n-6/n-3 ratios decreased. Thus, the addition of flaxseed oil to the tilapia diet led to an improvement in the quality of the fish meat in terms of physiological and nutritional properties.     

Fast Transmethylation of Total Lipids in Dried Blood by Microwave Irradiation and its Application to a Population Study

A methodology combining finger-pricked blood sampling, microwave accelerated fatty acid assay, fast gas chromatography data acquisition, and automated data processing was developed, evaluated and applied to a population study. Finger-pricked blood was collected on filter paper previously impregnated with 0.05 mg of the antioxidant butylated hydroxytoluene and air-dried at room temperature. Transmethylation was accelerated by microwave irradiation in an explosion-proof multimode microwave reaction system. The chemical procedure was based on a one-step direct transmethylation procedure catalyzed by acetyl chloride. The short-term stability of PUFA in blood dried on filter paper and storage at room temperature was examined using venous blood. The recoveries ranged from 97 to 101 % for the categorized fatty acids as well as the ratios of n-6 to n-3 PUFA and the n-3 % highly unsaturated fatty acid. Specifically, recoveries were 99, 98, 97, and 97 % for linoleic acid (18:2n-6), arachidonic acid (ARA), α-linolenic acid (ALA), and docosahexaenoic acid (DHA), respectively. The mol% (mean ± SD, 95 % confidence interval) of fatty acid composition in subjects from the population study was determined as 36.2 ± 3.8 (35.8, 36.7), 23.2 ± 3.0 (22.8, 23.5), 36.8 ± 3.5 (36.4, 37.2) and 3.79 ± 1.0 (3.68, 3.91) for the saturated, monounsaturated, n-6 and n-3 PUFA, respectively. Individually, the mean mol% (95 % CI) was 22.6 (22.3, 22.9) for 18:2n-6, 9.5 (9.3, 9.7) for ARA, 0.51 (0.49, 0.53) for ALA, 0.42 (0.38, 0.47) for eicosapentaenoic acid (EPA), and 1.67 (1.61, 1.73) for DHA. This methodology provides an accelerated yet high-efficiency, chemically safe, and temperature-controlled transmethylation, with diverse laboratory applications including population studies.     

Preparation of Highly Pure n-3 PUFA-Enriched Triacylglycerols by Two-Step Enzymatic Reactions Combined with Molecular Distillation

Highly pure n-3 polyunsaturated fatty acids (PUFA)-enriched triacylglycerols (TAG) have attracted considerable attention due to their nutritional benefits and pharmacological effects. In this study, an alternative approach to the conventional method for the synthesis of highly pure n-3 PUFA-enriched TAG by using a multi-step process was reported. First, glyceride mixtures were synthesized through Novozym 435 [Novozymes A/S (Bagsvaerd, Denmark)] catalyzed esterification of n-3 PUFA-enriched FA and glycerol. Second, partial glycerides in the resulting glyceride mixtures were hydrolyzed to FA by immobilized partial glycerides-selective lipase from Malassezia globosa. The purity of TAG reached 99.84% under the optimized conditions: buffer solution of pH 6.0, water content of 100% (w/w, with respect to the oil mass), enzyme loading of 120 U/g (U/w, with respect to oil mass) and reaction temperature of 30 °C. During hydrolysis, the immobilized SMG1-F278N exhibited good reusability and TAG purity of over 94% was maintained after being used for six cycles. Subsequently, purification of TAG was accomplished by molecular distillation at low temperature (150 °C) and highly pure (99.85%) TAG with 88.73% n-3 PUFA was obtained. The final glyceride mixtures with low acid, peroxide and anisidine value were promising products for medical and dietetic purposes. Compared with the conventional one-step synthesis of n-3 PUFA-enriched TAG by enzymatic esterification or glycerolysis or the two-step method by combined transesterification and ethanolysis, this improved process allows for higher purity of n-3 PUFA-enriched TAG and significant reduction in reaction time.     

Characteristics of the Fatty Acid Composition of a Deep-Sea Vent Gastropod, <Emphasis Type="Italic">Ifremeria nautilei</Emphasis>

Neutral and polar lipids in the soft parts of a gastropod species, Ifremeria nautilei, collected from deep-sea hydrothermal vents, were examined to assess the trophic relationships in hydrothermal vents. The vent gastropod obtains many of its lipids from symbiotic chemosynthetic microorganisms. The major polyunsaturated fatty acids (PUFA) both in the triacylglycerols and phospholipids of the gastropod consist of a limited number of n-3 and n-6 PUFA: arachidonic acid (20:4n-6), icosapentaenoic acid (20:5n-3), and docosapentaenoic acid (22:5n-3), without docosahexaenoic acid (DHA, 22:6n-3). Noticeable levels of various n-6 PUFA, such as 18:2n-6,9, 20:2n-6,9, 20:3n-6,9,12, and 20:3n-6,9,15 with significant levels of 16:1n-6 and 18:1n-6 indicate the biosynthetic characteristic of the endosymbionts. The lack of DHA in all specimens suggests a limitation of its lipid biosynthesis ability with its symbionts. This finding with regard to the lipids is unusual for a marine animal in the grazing or detrital food chain because many marine animal lipids evidently contain high levels of DHA with low levels of n-6 fatty acids. Such contradictory findings lead to some new insights into the absence of a biosynthetic pathway for DHA in I. nautilei, and provide evidence that DHA in this species is dispensable. Similar to herbivorous gastropods, the lack of DHA with significant levels of n-6 PUFA in this species also indicates its selective assimilation of specific microorganisms, such as chemosynthetic bacteria in hydrothermal vents, because significant levels of DHA were found in carnivorous mollusk lipids.     

Differences in Lipid Characteristics Among Populations: Low-Temperature Adaptability of Ayu, Plecoglossus altivelis

The lipid and fatty acid compositions of the total lipids of three cultured populations (migratory between fresh and salt water, Lake Biwa landlocked, and Setogawa River forms) of ayu, Plecoglossus altivelis, were investigated to clarify the difference in lipid characteristics and temperature adaptability among the three groups. Triacylglycerols were the dominant depot lipids of the three populations, while phospholipids, such as phosphatidylcholine and phosphatidylethanolamine, were found to be the major components of the polar lipids, and their lipid classes are similar to each other. The major fatty acids in the triacylglycerols of all specimens were 16:0, 18:0, 16:1n-7, 18:1n-7, 18:1n-9, 18:2n-6 (linoleic acid), 20:5n-3 (EPA, icosapentaenoic acid), and 22:6n-3 (DHA, docosahexaenoic acid), similar to the tissue phospholipids of the three populations, 16:0, 18:0, 16:1n-7, 18:1n-7, 18:1n-9, 18:2n-6, 20:4n-6, EPA, and DHA. All classes had high levels of 18:2n-6, which originates from their dietary lipids. Compared with the lower DHA levels of the triacylglycerols, the higher levels in the phospholipids suggest their selective accumulation or a biosynthetic pathway to DHA as in freshwater fish. Two populations (the migratory and Setogawa River forms) adapted to lower temperatures with comparatively high levels of polyunsaturated fatty acids (PUFA) for their membrane fluidities. With significantly higher levels of n-3 PUFA and total PUFA, the mean DHA content in the lipids of the Setogawa River form (the population that adapted to lower temperatures) was significantly higher than that of the migratory form. From these results, we concluded that the Setogawa River population actively concentrates long-chain PUFA in its polar lipids and has high adaptability to low temperature.