Characterization of lecithin isolated from anchovy (Engraulis japonica) residues deoiled by supercritical carbon dioxide and organic solvent extraction

Lecithin was isolated and characterized from anchovy (Engraulis japonica) deoiled residues using supercritical carbon dioxide (SC-CO(2)) at a semibatch flow extraction process and an organic solvent (hexane) extraction. SC-CO(2) extraction was carried out to extract oil from anchovy at different temperatures (35 to 45 °C) and pressures (15 to 25 MPa). Extraction yield of oil was influenced by physical properties of SC-CO(2) with temperature and pressure changes. The major phospholipids of anchovy lecithin were quantitatively analyzed by high-performance liquid chromatography. Phosphatidylcholine (PC) (68%± 1.00%) and phosphatidylethanolamine (PE) (29%± 0.50%) were the main phospholipids. Thin layer chromatography was performed to purify the individual phospholipids. The fatty acid compositions of lecithin, PC, and PE were analyzed by gas chromatography. A significant amount of eicosapentaenoic acid and docosahexaenoic acid were present in both phospholipids of PC and PE. Emulsions of lecithin in water were prepared through the use of a homogenizer. Oxidative stability of anchovy lecithin was high in spite of its high concentration of long-chain polyunsaturated fatty acids. PRACTICAL APPLICATION: Lecithin can be totally metabolized by humans, so is well tolerated by humans and nontoxic when ingested. Lecithin from anchovy contain higher amounts of ω-3 fatty acids especially EPA and DHA, it may have positive outcome to use in food and pharmaceutical industries.     

草鱼头磷脂制备工艺优化及抗氧化性能分析

为了得到较高纯度的草鱼头磷脂并分析其抗氧化性能,采用乙醇溶液浸提冷冻干燥草鱼头粉末制备磷脂,正交试验确定其最优工艺后,利用薄层层析(TLC)分析比较草鱼头磷脂种类的组成及含量,利用气相色谱-质谱联用技术分析磷脂的脂肪酸组成,并进行体外抗氧化能力测定。结果表明:磷脂制备最优工艺方案为:乙醇浓度80%,料液比1:6 g/mL,提取温度65℃,提取时间3 h,在此条件下,得到草鱼头磷脂的纯度为84.20%±0.65%,提取率为3.03%±0.06%。草鱼头磷脂主要含有磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、鞘磷脂(SM)、溶血磷脂酰胆碱(LPC)四种磷脂,其中PC的含量最高(60.00%±0.26%),其次为PE(20.00%±0.17%)。草鱼头磷脂脂肪酸组成以多不饱和脂肪酸(PUFA)为主(52.09%±0.59%),其中,C20:5n-3(EPA)和C22:6n-3(DHA)含量较高。体外抗氧化测定试验结果表明,草鱼头磷脂的羟自由基(·OH)清除能力和还原力能力都显著(P<0.05)优于对照的商品大豆磷脂。     

六种鱼鱼头磷脂的组成及脂质脂肪酸分析

为研究海水鱼和淡水鱼鱼头磷脂及脂质脂肪酸组成的差异,以海水鱼(马鲛鱼、巴浪鱼、金鲳鱼)和淡水鱼(罗非鱼、鲫鱼、鲢鱼)鱼头为研究对象,利用Floch法提取6种鱼鱼头总脂;硅胶柱层析法将总脂分离收集中性脂、糖脂和磷脂;薄层层析(thin layer chromatography,TLC)分析比较6种鱼鱼头磷脂种类的分布、组成;气相色谱(Gas chromatography,GC)分析6种鱼鱼头脂质的脂肪酸组成。结果表明,6种鱼鱼头中提取率最高的脂质类型为中性脂,占总脂的48.98%~77.84%,其次为磷脂,占总脂的5.93%~22.86%,糖脂提取率最低,占总脂的3.67%~15.91%。鱼头磷脂中共检出4种磷脂组分,分别为磷脂酰乙醇胺(Phosphatidylethanolamine,PE)、磷脂酰胆碱(Phosphatidylcholine,PC)、鞘磷脂(Sphingomyelin,SM)、溶血磷脂酰胆碱(Lysophosphatidylcholine,LPC),其中PC明显高于其他磷脂组分。脂质中均含有较高含量的多不饱和脂肪酸(Poly-unsaturated fatty acid,PUFA),其中以二十二碳六烯酸(docosahexaenoic acid,DHA)为主,且均含有二十碳五烯酸(eicosapntemacnioc acid,EPA),海水鱼和淡水鱼鱼头磷脂中DHA和EPA占总脂肪酸的比例最高,分别为18.39%~21.43%和5.61%~10.38%,且海水鱼鱼头中以DHA和EPA为主的PUFA含量高于淡水鱼鱼头。因此,海水鱼鱼头是提取n-3多不饱和脂肪酸的潜在资源,尤其是制备n-3多不饱和脂肪酸型磷脂的良好来源。     

Supercritical fluid extraction and characterization of lipids from algae Scenedesmus obliquus

Supercritical carbon dioxide (SC-CO2) extractions (with and without ethanol as an entrainer) were carried out to remove lipids and pigments from protein concentrate of green algae (Scenedesmus obliquus) cultivated under controlled conditions. The content and fatty acid composition of algal lipids using column, thin-layer (TLC) and gas-liquid chromatography (GLC) were determined. Absorption spectra of extracted fractions showed the predominance of chlorophyll A (lambda max at 410 nm). Single step supercritical carbon dioxide (SC-CO2) extraction resulted mostly in removal of neutral lipids and a part of glycolipids, but phospholipids were not extracted. Addition of ethanol to SC-CO2 increased the amount of glycolipids and phospholipids in the extract. TLC pattern of algal lipids showed that the main part of neutral lipids consisted of diglycerides, triglycerides, hydrocarbons, free sterols, and sterol esters. The glycolipids were mostly monogalactosyl diglyceride, digalactosyl diglyceride, esterified sterol glycoside, and sterol glycoside. In phospholipids, phosphatidyl choline, phosphatidyl glycerol, and phosphatidyl ethanolamine were the main compounds. Fatty acid composition patterns indicated the main fatty acids to be 16:0, 16:1, 16:2, 16:3, 16:4, 18:1, 18:2, and 18:3(a). Relatively high recovery of polyunsaturated fatty acids and essential fatty acids in supercritical fluid extracted algal lipids and proteins isolates were observed.     

Determination of intramuscular phospholipid classes and molecular species in Gaoyou duck

In this study, intramuscular phospholipid classes and molecular species in Gaoyou duck meat were determined. Classes of phospholipids were identified and quantified by normal phase HPLC combined with UV and evaporative light scattering detectors (ELSD). The main phospholipid classes (phosphatidylcholine and phosphatidylethanolamine) were prepared on a semi-preparative silica gel column by HPLC. Reverse phase HPLC was coupled in parallel with both an ELSD and a mass spectrometry in order to characterise molecular species of phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The results showed that Gaoyou duck meat had high quantities of PC and PE (64.66% and 28.10% of total phospholipids, respectively). Arachidonic acid was mainly present in PE and formed molecular species containing a saturated fatty acid, such as stearic or myristic acid; however, oleic acid together with palmitic or stearic acid formed the main molecular species in PC. The content of the molecular species with polyunsaturated fatty acids in PE accounted for 98.33%, while that in PC only 46.20%.     

鲢鱼各部位磷脂组分及脂肪酸组成分析

应用Bligh-Dyer法、硅胶柱层析和薄层层析（thin layer chromatography,TLC）分析比较鲢鱼头部、眼部、背肌、腹肌、内脏、尾部的磷脂含量与组成,气相色谱-质谱联用技术分析鲢鱼各部位磷脂的脂肪酸组成。结果表明：各部位磷脂种类有磷脂酰胆碱（phosphatidylcholine,PC）、磷脂酰乙醇胺（phosphatidylethanolamine,PE）、磷脂酰丝氨酸（phosphatidyelserine,PS）及一未知组分,鲢鱼各部位的PE、PC明显高于其他磷脂组分。鱼头磷脂中饱和脂肪酸的总量最高,其次是内脏中含量,鱼眼中含量最低。单不饱和脂肪酸的含量也在鱼头中最丰富,明显高于背肌、鱼眼、腹肌、鱼尾,后三者间差异性不显著（P＞0.05）,内脏中含量最低。多不饱和脂肪酸的含量在鱼眼中最多,与腹肌、鱼尾中的含量差异不显著,都在49%以上,但明显高于鱼头、背肌内脏中的含量。各部位间内脏中二十碳五烯酸和二十二碳六烯酸的含量明显高于其他部位,其次是鱼眼。鱼眼磷脂中C22∶6的含量最丰富,C20∶5在内脏磷脂中含量最多。     

Determination of phospholipid molecular species in pork meat by high performance liquid chromatography-tandem mass spectrometry and evaporative light scattering detection

Normal phase high performance liquid chromatography has been optimized for both evaporative light scattering detection and tandem mass spectrometry in order to characterize the natural phospholipids (PL) (classes and molecular species) of raw and cooked pork meat. The PL fraction included phosphatidylcholine (PC) (42.9%±4.5 for raw vs 42.6%±8.0 for cooked meat), plasmalogen-phosphatidylethanolamine (pPE) and phosphatidylethanolamine (PE) (26.7%±3.1 vs 28.5%±2.3), cardiolipin (CL) (8.3%±2.9 vs 6.3%±0.7), sphingomyelin (Sph) (7.5%±0.9 vs 8.3%±2.1), phosphatidylinositol (PI) (6.8%±0.7 vs 6.5%±2.1) phosphatidylserine (PS) (4.9%±0.5 vs 4.6%±1.4) and lysophosphatidylcholine (LPC) (2.9%±1.3 vs 3.3%±2.6). Arachidonic acid (absent in Sph) was mainly present in pPE and PI and formed molecular species with a saturated fatty acid, such as stearic (as in PI, PS, PE and PC) or palmitic acid (as in PE and PC), or the respective vinyl ethers in pPE (p18:0 and p16:0); however, in PC, arachidonic acid also formed combinations with oleic and linoleic acid. Palmitic acid formed the most abundant molecular species in PC, but not in CL, PE, PI and PS. Unexpectedly, the cooked pork meat showed an increased content of the molecular species of PI and LPC with more unsaturated fatty acids (18:0/20:4 and 18:2, respectively) with respect to raw meat.     

金华火腿加工过程中肌肉肌内脂的水解变化研究

研究了金华火腿股二头肌 (bicepsfemoris)肌内脂肪在加工过程中不同时期的水解变化情况。肌内脂使用氯仿 甲醇溶液提取 ,采用固相萃取法将中性脂 (主要为甘油脂 )、游离脂肪酸和磷脂分离 ,用毛细管气相色谱分别分析甘油脂、游离脂肪酸和磷脂脂肪酸的种类与比例。通过比较不同时期样品的色谱结果 ,发现在金华火腿原料中磷脂脂肪酸的不饱和脂肪酸百分含量较高 ,尤其是多不饱和脂肪酸的含量高达 4 9 2 2 %。且在金华火腿成熟过程中 ,磷脂水解作用显著 ,而甘油脂脂肪酸百分比例比较稳定 ,说明甘油脂水解作用相对较弱。     

鲟鱼卵脂质的提取及脂肪酸组成分析

以西伯利亚鲟鱼卵为原料,比较不同提取方法对鲟鱼卵脂质提取率、脂肪酸及脂质组成的影响。采用索氏提取、酶辅助有机溶剂和超临界CO2三种不同的方法提取鲟鱼卵中的脂质,并分析鲟鱼卵脂肪酸组成及脂质组成。结果显示,不同提取方法对所得到的鲟鱼卵脂质的提取率和脂肪酸的相对含量有显著的影响（P＜0.05）,对鲟鱼卵脂质的脂肪酸种类和脂质组成没有显著影响（P＞0.05）。索氏提取法的提取率为（23.71±1.82）%,中性蛋白酶酶解辅助有机溶剂法的提取率为（15.47±1.21）%,超临界CO2萃取法的提取率为（10.43±2.16）%。鲟鱼卵脂质中含有17 种脂肪酸,包括6 种饱和脂肪酸,4 种单不饱和脂肪酸,7 种多不饱和脂肪酸,其中总不饱和脂肪酸含量达到70%以上,多不饱和脂肪酸含量达到17%以上。不同提取方法提取鲟鱼卵脂质中均含有甘油三酯、胆固醇、极性脂质,其中甘油三酯相对含量达到89%以上。     

秘鲁鱿鱼不同组织营养成分分析与评价

为实现秘鲁鱿鱼资源的高值化利用,分析秘鲁鱿鱼不同组织(头部、肌肉、内脏、眼部和颈部)的基本营养组成、氨基酸组成、脂肪酸组成和矿物质元素含量,比较其不同组织的营养特性。结果表明,秘鲁鱿鱼不同组织之间营养组成存在显著差异。不同组织水分含量均在75%以上,眼部水分含量最高(84.85%);肌肉的粗蛋白含量最高(18.21%)且粗脂肪含量最低(0.33%);内脏的粗脂肪(2.75%)和总糖(0.60%)含量均显著高于其它组织。所有组织均含有18种氨基酸,且不同组织的必需氨基酸含量占比均大于36%。每种必需氨基酸的氨基酸评分(amino acid score,AAS)大多数在0.60以上,化学评分(chemical score,CS)大多数低于鸡蛋蛋白模式。不同组织分别检测出8种~11种脂肪酸,不饱和脂肪酸含量均大于64%,眼部的多不饱和脂肪酸含量最高(76.30%),肌肉组织的二十二碳六烯酸(docosahexaenoic acid,DHA)含量最高(46.83%),颈部的二十碳五烯酸(eicosapentaenoic Acid,EPA)和DHA占比最高(61.49%)。所有组织中常量元素K、Na和P含量最为丰富,内脏中Mg、Ca、Cu、Fe、Zn、Mn、Se的含量远高于其它组织。     

水酶法提取斑点叉尾鮰内脏油的工艺研究

为提高鱼类加工废弃物的综合利用率,以斑点叉尾鮰内脏为原料,采用水酶法制备鱼油。结果表明：水酶法提取鱼油的最佳工艺条件为初始pH7.5、料液比1:1、加酶量3500U/g、酶解温度55℃、酶解时间0.5h,该条件下鱼油的提取率达到80.94%;提取到的鱼油(即粗鱼油)外观呈黄褐色,具有鱼腥味和轻微的酸败味,其过氧化值较低,而酸值相对较高;粗鱼油中的饱和脂肪酸相对含量为19.42%,单不饱和脂肪酸和多不饱和脂肪酸分别为61.76% 和16.83%,Σn-3/Σn-6 的脂肪酸比值达到3.63。表明斑点叉尾鮰内脏是生产鱼油的良好来源。     

基于1,2-二氯乙烷体系的金枪鱼副产物中脂质提取与组学分析

为提高金枪鱼副产物的利用率,减少资源浪费,本试验基于1,2-二氯乙烷体系结合丙酮沉淀法,建立从金枪鱼副产物中同时提取脂质和鱼油的新方法.采用气相色谱法和液相色谱-质谱联用法分别对提取脂质中鱼油和磷脂进行脂质组学研究.通过调节溶剂体系得到1,2-二氯乙烷-甲醇的最佳体积比为1:2.进一步优化提取条件,得到磷脂和鱼油的得率...     

Short communication: Effects of dietary deoiled soy lecithin supplementation on circulating choline and choline metabolites,and the plasma phospholipid profile in Holstein cows fed palm fat

Dietary lecithin is a source of choline. Our objective was to evaluate the effects of dietary deoiled soy lecithin feeding on circulating choline, choline metabolites, and the plasma phospholipid profile in lactating dairy cows fed fractionated palm fatty acids. In a split-plot Latin square design, 16 Holstein cows (160 ± 7 d in milk; 3.6 ± 1.2 parity) were randomly allocated to a main plot receiving a corn silage and alfalfa haylage-based diet with palm fat containing either moderate or high palmitic acid content at 1.75% of ration dry matter (moderate and high palmitic acid containing 72 or 99% palmitic acid in fat supplement, respectively; n = 8/palm fat diet). Within each palm fat group, deoiled soy lecithin was top-dressed at 0, 0.12, 0.24, or 0.36% of ration dry matter in a replicated 4 × 4 Latin square design with 14-d experimental periods. A 14-d covariate period was used to acclimate cows to palm fat feeding without lecithin supplementation. Blood sampling occurred during the final 3 d of each experimental period. Plasma choline and choline metabolites were quantified using liquid chromatography and mass spectrometry. Plasma phospholipids were profiled using time-of-flight mass spectrometry. Whereas no effects of treatments were detected for plasma choline or methionine, lecithin feeding increased the plasma concentrations of choline metabolites trimethylamine N-oxide and dimethylglycine (24 and 11%, respectively). Plasma phosphatidylcholine (PC) and sphingomyelin (SM) concentrations increased with deoiled lecithin feeding (e.g., PC 16:0/22:6 and SM d18:1/18:3). Lecithin supplementation also increased plasma lysophosphatidylcholine (LPC) concentrations (e.g., LPC 18:0) while reducing plasma phosphatidylethanolamine (PE) concentrations (e.g., PE 16:0/20:5). Although increases in microbial-derived trimethylamine N-oxide suggest gastrointestinal lecithin degradation, elevations in plasma dimethylglycine, PC, LPC, and SM suggest that choline availability was improved by lecithin feeding in cows, thus supporting enhanced endogenous phospholipid synthesis.     

微波辅助酶法提取青鱼内脏鱼油工艺优化及脂肪酸组成分析

以青鱼内脏为原料,采用微波辅助蛋白酶提取鱼油,研究酶的种类、酶添加量、酶解时间、酶解温度、微波功率、微波处理时间等因素对鱼油提取得率的影响。采用Box-Behnken响应曲面设计法,建立影响因素和鱼油得率之间的回归方程。结果表明:中性蛋白酶为最佳水解酶,微波功率400 W、酶加入量2%、酶解时间2.5 h、酶解温度45℃、微波处理时间15 min为鱼油最佳提取条件,在此条件下鱼油得率为26.26%±0.13%。使用气相色谱法对萃取的青鱼内脏鱼油脂肪酸组成进行定性和定量分析,结果显示饱和脂肪酸有3种,占总脂肪酸含量的22.39%,主要为棕榈酸和硬脂酸;单不饱和脂肪酸有3种,占总脂肪酸含量的40.42%,主要为油酸和棕榈油酸;多不饱和脂肪酸有8种,占总脂肪酸含量的37.19%,主要为亚油酸。     

Influence of dietary fish meal on egg fatty acid composition

Changes in the fatty acid composition of egg-yolk fat of hens fed diets with increasing fish meal content are studied by total fatty acid analysis. The fatty acid composition of the major lipid fractions in egg-yolk fat after feeding a high-level fish meal diet was determined by a combination of thin layer chromatography (t.l.c.) and gas-liquid chromatography (g.l.c.) The changes produced show a fatty acid pattern similar to those of the diets themselves. Long-chain polyunsaturated fatty acids are deposited preferentially in the phospholipids, reaching the highest concentration in cephalin and lecithin.     

褐点石斑鱼鱼肉及其副产物的脂肪酸分析

为了评估褐点石斑鱼的脂肪酸组成特点,采用固相萃取和气相色谱技术对鱼肉、头、皮和内脏的脂质含量、总脂及各脂类脂肪酸组成,甘油三酯(triacylglycerols, TAG)和磷脂(phospholipids, PL)中脂肪酸的位置分布进行了较全面的分析。石斑鱼鱼头和内脏中含有丰富的糖脂(saccharolipids, SL)和PL。在各组织总脂中,必需脂肪酸的含量占总脂肪酸的24.17%～24.51%,二十二碳六烯酸(docosahexaenoic acid, DHA)及二十碳五烯酸(eicosapentaenoic acid, EPA)的含量为12.42%～13.73%,动脉粥状硬化指数、血栓形成指数和n-6/n-3的比例处于较低水平。多不饱和脂肪酸(polyunsaturated fatty acids, PUFA),尤其是DHA+EPA(28.43%～37.37%)在PL中的比例要远高于在其它脂类中的占比。饱和脂肪酸主要分布在TAG和PL的sn-2位,单不饱和脂肪酸主要分布在TAG的sn-1,3位和PL的sn-2位,PUFA倾向于分布在TAG的sn-1,3位和PL的sn-1位;T...     

Changes in lipid composition,fatty acid profile and lipid oxidative stability during Cantonese sausage processing

Lipid composition, fatty acid profile and lipid oxidative stability were evaluated during Cantonese sausage processing. Free fatty acids increased with concomitant decrease of phospholipids. Total content of free fatty acids at 72 h in muscle and adipose tissue was 7.341 mg/g and 3.067 mg/g, respectively. Total amount of saturated, monounsaturated and polyunsaturated fatty acids (SFA, MUFA, and PUFA) in neutral lipid exhibited a little change during processing, while the proportion of PUFA significantly decreased in the PL fraction. The main triacylglycerols were POO + SLO + OOO, PSO (P = palmitic acid, O = oleic acid, L = linoleic acid, S = stearic acid), and a preferential hydrolysis of palmitic, oleic and linoleic acid was observed. Phosphatidylcholines (PC) and phosphatidylethanolamines (PE) were the main components of phospholipids and PE exhibited the most significant degradation during processing. Thiobarbituric acid values (TBARS) increased while peroxide values and hexanal contents varied during processing.     

Lipid Content and Fatty Acid Composition in Bamboo Shoots

Lipids from bamboo shoots (Phyllostachys pubescens), peeled and divided from top to base, were extracted and fractionated into three classes, and each class separated into constituent components by thin-layer chromatography (TLC). Fatty acid composition and amount of separated lipids were determined. Total lipids (TL) ranged from 800 (top) to 380 mg (base) per 100g fresh weight and the ratio of nonpolar lipids (NPL):glycolipids (GL):phospholipids (PL) was about 17:27:56. The main fatty acids of the three lipid classes were palmitic, linoleic and linolenic acids, but composition was remarkably different among these fractions. The fatty acid composition of triglycerides (TG) was similar to the original NPL. Palmitic acid was almost all located in 1-, 3-position, linoleic acid mainly located in 2-position of TG, while linolenic acid was distributed in each position. Digalactosyl diglyceride (DGDG) and monogalactosyl diglyceride (MGDG) were the main components of GL; the average of the former had about 37% linoleic and 29% linolenic acids, while the latter had about 25% linoleic and 62% linolenic acids. Bamboo shoots contained 9 PL fractions, the major being phosphatidyl choline (PC) and phosphatidyl ethanolamine (PE). PC contained about 48% linoleic, 31% palmitic and 11% linolenic acids, and PE also had the similar tendency as PC.     

鱿鱼内脏粗提油的精制工艺优化及其理化指标分析

本研究采用碱炼法脱酸、活性白土分批脱色对鱿鱼内脏粗提油进行精炼。分别以酸价和脱色率作为脱酸和脱色效果的评价指标,在单因素实验的基础上,运用中心组合设计法优化精制工艺,同时分析测定了精制前后鱿鱼内脏油的相关理化指标。结果表明鱿鱼内脏粗提油的最佳脱酸工艺为:碱液浓度26 °Bé、碱炼温度54℃、碱炼时间26 min,在此脱酸条件下鱿鱼内脏油的酸价为(0.63±0.05)mg/g;最佳脱色工艺为:活性白土添加量3.7%、脱色温度88℃、脱色时间60 min,在此脱色条件下鱿鱼内脏油的脱色率为92.38%±1.25%。精制后鱿鱼内脏油的酸价和色泽达到一级精制鱼油的质量标准,其他各项理化指标达到或接近二级精制鱼油的质量标准。精制后鱿鱼内脏油的多不饱和脂肪酸含量达38.91%,其中EPA和DHA含量分别为12.56%和16.32%。以上结果说明本研究优化的工艺能够对鱿鱼内脏粗提油进行有效的精炼,可用于工业化生产。     

Original article: Extraction of lipid from scallop (Patinopecten yessoensis) viscera by enzyme‐assisted solvent and supercritical carbon dioxide methods

In the present study, lipid was extracted from scallop (Patinopecten yessoensis) viscera by using the enzyme‐assisted solvent method and the supercritical carbon dioxide (SC‐CO₂) method. Soxhlet extraction with ethyl ether produced a yield of 23.7 ± 0.6 g of lipid 100 g^?1 of dry matter. Enzyme‐assisted solvent extraction allowed recovering 60.6 ± 1.5% of P. yessoensis viscera lipid from the samples treated with papain, whereas a lipid recovery rate of 78.3 ± 0.6% was achieved by SC‐CO₂ extraction. The lipid extracted was divided into the unsaponifiable fraction (sterol) and the saponifiable fraction (fatty acid) and analysed by gas chromatography mass spectrometry. Results indicated that the fatty acid composition and sterol composition for lipids extracted by different methods were slightly different. Eicosapentaenoic acid and docosahexaenoic acid were dominant polyunsaturated fatty acids accounting for 35–40% of the total fatty acid.