虾头中磷脂提取与组学分析

研究虾头中磷脂的提取制备方法,并分别利用气相色谱法和多维度串联质谱法对样品磷脂的脂肪酸链结构进行脂质组学分析。利用乙醇浸提法制备得到虾头中的磷脂,并从乙醇体积分数、提取温度和提取时间3?个因素对提取方法进行优化,得到较优条件为乙醇体积分数90%、提取温度50?℃、提取时间30?min,磷脂的提取效果最佳,实际提取量可达到（11.58±0.03）mg/g,与优化前相比提高了83.8%。磷脂样品经甲酯化反应后用气相色谱法分析其脂肪酸链组成,并用高效液相色谱-质谱联用技术经负离子全扫描对磷脂分子实现分离鉴定和定量分析。结果显示,磷脂的脂肪酸链中主要含有棕榈酸链、亚油酸链、二十碳五烯酸和硬脂酸链等23?种脂肪酸链,其中单不饱和脂肪酸链占9.51%,多不饱和脂肪酸链占35.33%;磷脂样品中共检出磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰肌醇和磷脂酰丝氨酸4?类共28?种磷脂分子,该提取方法所得各种类磷脂的脂肪酸链中存在许多不饱和度较高的脂肪酸链,如40∶8、36∶7、38∶7、O-40∶7、40∶7、34∶6、O-36∶6、36∶6、O-38∶6、38∶6、40∶6等。虾头中磷脂含量丰富,其脂肪酸链不饱和度较高,因此具有较高的开发利用价值。     

羊骨脂质的组成分析

为了实现羊骨副产物的有效利用和为羊骨油高值化产品开发提供基础数据，以羊腿骨为原料，提取其中的油脂，分别采用高效液相色谱法(HPLC)、多维质谱鸟枪法(MDMS-SL)、气相色谱-质谱联用法(GC-MS)对羊骨油的甘油酯组成、脂质组学、脂肪酸组成进行分析。结果表明：羊骨油中甘油三酯含量达到96.76%,还有少量的甘油二酯(1.41%),而甘油一酯未检出；羊骨油中共鉴定出19种酰基肉碱、17种鞘磷脂、37种磷脂酰胆碱、39种甘油三酯，总含量分别为5.76、96.58、85.18 nmol/g和15.76μmol/g;羊骨油中共鉴定出11种脂肪酸，其中饱和脂肪酸含量为11.51%,单不饱和脂肪酸含量为72.34%,多不饱和脂肪酸含量为7.56%。     

超高效液相色谱-静电场轨道阱高分辨质谱鉴定核桃仁的脂质构成

利用超高效液相色谱-静电场轨道阱高分辨质谱系统鉴定了核桃的脂质组。在正负离子模式下获得脂质的一级质谱和二级质谱信息后,共鉴定出4 大类525 种脂质分子,包括250 种甘油酯、221 种磷脂、18 种糖脂、36 种鞘脂类,含量分别占总脂质的87%、12.97%、0.02%和0.01%。甘油二酯（diacylglycerol,DG）和甘油三酯（triacylglycerol,TG）是甘油酯的主要成分,分别占总脂质的45.23%和41.77%;TG分子中以TG(18:2/18:2/18:3)含量最高,DG分子中DG(18:2/18:2)含量最高,亚油酸和亚麻酸等人体必需脂肪酸是DG和TG中的主要脂肪酸。核桃中磷脂种类丰富,包括磷脂酸、磷脂酰甘油、磷脂酰丝氨酸、磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰肌醇、磷脂酰肌醇二磷酸、溶血性磷脂酰胆碱、溶血性磷脂酰乙醇胺、溶血性磷脂酰甘油、溶血性磷脂酰肌醇和双磷脂酰甘油,其中磷脂酸最多,约占磷脂的43.88%。本实验构建的核桃脂质图谱,可为核桃功能开发和营养品质的深入研究提供一定理论依据。     

乙酯型金枪鱼油不同级分子蒸馏过程中脂肪酸含量及挥发性物质变化

乙酯型金枪鱼油是粗鱼油经过多级分子蒸馏后得到的富含多不饱和脂肪酸,无色无味的优质鱼油,在这个过程中发生了一系列变化。利用气相色谱-质谱联用仪、电子鼻、傅里叶变换红外光谱仪检测不同蒸馏组分的金枪鱼油在成分、风味和性质上的变化以及主要的挥发性物质。结果表明:原料鱼油和不同蒸馏组分鱼油中二十二碳六烯酸(docosahexaenoic acid,DHA)和二十碳五烯酸(eicosapentaenoic acid,EPA)的含量呈先增加后下降趋势,其中EPA含量可达31.41%,DHA含量可达74.10%;二级分子蒸馏可以得到DHA、EPA总含量高达84.26%的乙酯型鱼油,而三级、四级、五级蒸馏鱼油中DHA和EPA含量显著下降。电子鼻检测原料鱼油及不同蒸馏组分的鱼油挥发性物质差异显著。不同蒸馏组分的金枪鱼油主体风味物质为己醛、辛醛、庚醛、壬醛、1-辛烯-3-醇、2-十一酮、2-戊基呋喃等,共同赋予其鱼腥、油脂气息、泥土味、脂肪香味、青草味、蘑菇香、蔬菜香味等。红外光谱分析说明了不同级分子蒸馏后得到的金枪鱼油的主要成分含量是有所不同的。不同蒸馏组分的鱼油中一级、二级鱼油脂类成分种类最多,富集效果最好,而三级、四级、五级分子蒸馏富集效果显著下降。     

酶辅助提取南极磷虾磷脂及脂质组学研究

南极磷虾中含有丰富的磷脂资源,是磷脂的生物富集库。采用蛋白酶辅助Bligh & Dyer法提取南极磷虾磷脂技术,研究温度、pH值、时间以及酶的种类对南极磷虾磷脂提取率的影响;采用气相色谱法研究磷脂脂肪酸链结构组成,建立亲水色谱串联质谱法（HILIC-QTOF/MS）法对磷脂进行定量和鉴定。研究发现胰蛋白酶为最优提取酶,通过Box-Benhnken方法优化提取南极磷虾磷脂的最佳条件为：提取温度41 ℃,pH 9,提取时间3 h。该条件下得到的磷脂提取量为（3.66 ± 0.03）%。经测定,本试验中提取的南极磷虾磷脂的脂肪酸链结构主要为C16:0（31.58%）、EPA（31.94%）、DHA（20.64%）等。采用酶辅助Bligh & Dyer法提取得到的脂肪酸中不饱和脂肪酸所占比例比较高,EPA和DHA达到52.58%。通过HILIC-QTOF/MS 脂质组学研究成功分离磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）和磷脂酰肌醇（PI）三大类磷脂。对磷脂类中各磷脂分子实现离子源内分离后成功测定59种磷脂分子,经数据库比对和二级质谱佐证得到磷脂脂肪酸链结构信息：南极磷虾中存在大量含有EPA和DHA链的磷脂分子。本研究结果为南极磷虾磷脂的开发和综合利用提供了试验依据。     

GC-MS分析新疆拜城油鸡和南京土鸡鸡皮脂质中脂肪酸组成

分析比较新疆拜城油鸡和南京土鸡鸡皮总脂质和磷脂的含量及脂肪酸组成。用溶剂提取法分别提取2 种鸡皮的总脂质和磷脂,将提取的脂质经甲酯化后,利用气相色谱-质谱分析其脂肪酸组成。结果显示,南京土鸡鸡皮总脂质含量和磷脂含量分别为35.56%和7.71%,拜城油鸡鸡皮中总脂质含量和磷脂含量分别为42.65%和8.95%,均高于南京土鸡。南京土鸡与拜城油鸡鸡皮总脂质中各含有25 种脂肪酸,主要包含油酸、亚油酸、棕榈酸、硬脂酸、棕榈烯酸;2 种鸡鸡皮磷脂中均含有10 种脂肪酸,主要包含棕榈酸、油酸、硬脂酸和亚油酸。2 种鸡的鸡皮油脂脂肪酸的种类相同,但各脂肪酸含量有所差异;脂肪酸饱和程度比例基本相似,但南京土鸡鸡皮磷脂中不饱和脂肪酸和亚油酸含量高于拜城油鸡。     

分子蒸馏法制备鱼油多不饱和脂肪酸

研究了多级分子蒸馏法提取深海鱼油中多不饱和脂肪酸的工艺方法,通过对压力和温度的控制得到不同多不饱和脂肪酸含量的各级鱼油产品,当蒸馏温度为110℃以上,蒸馏压力为20Pa以下时,经过三级串联分子蒸馏,得到高碳链不饱和脂肪酸质量分数为90.96%的鱼油产品,并用气相色谱法测定了产品的脂肪酸组成,分析了产品的理化指标.     

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。     

烟熏对鲟鱼肉特性及脂质组成的影响

为研究烟熏工艺对鲟鱼腹部与背部鱼肉组织特性的影响,采用亲水作用色谱-质谱联用(HILIC-MS)技术分析烟熏工艺对鲟鱼肉磷脂组成的影响,并通过主成分分析(PCA)方法研究磷脂组成差异性。研究表明:烟熏后鲟鱼肉具有更高的硬度、内聚性、弹性、胶黏性和咀嚼性,然而多不饱和脂肪酸含量下降。脂质组学方法检测出鲟鱼肉中磷脂酰胆碱(PC)分子20种,磷脂酰乙醇胺(PE)分子24种,磷脂酰丝氨酸(PS)分子12种。主成分分析表明磷脂可作为区分鲟鱼肉腹部和背部的定性指标,然而烟熏后鲟鱼肉背部与腹部磷脂组成差异性减少。本研究表明鲟鱼肉腹部因不饱和脂肪酸含量较高而不适用热熏工艺,应考虑液熏等冷熏加工工艺。     

三种国标方法测定酒中甜蜜素的比较与方法改进

通过比较气相色谱法、液相色谱法、液相色谱-质谱/质谱法3种国标方法测定酒中甜蜜素含量,发现国标气相色谱法不能将酒中的干扰物分离,常出现甜蜜素假阳性结果;液相色谱法虽无基质干扰现象,满足配制酒的检测要求,但样品衍生化时会产生有害物质,对操作者有潜在的危害;液相色谱-质谱/质谱法虽是酒中甜蜜素检测最佳方法,但设备比较昂贵,应用受限。为此对国标气相色谱法进行了改进,通过优化色谱条件,使改进后的气相色谱法能有效地消除基质干扰,其检出限、回收率及精密度均能满足配制酒的检测要求,且方法技术参数均优于液相色谱法,扩展了国标气相色谱法的适用范围。同时对国标液相色谱-质谱/质谱法进行了优化,优化后的方法检出限为0.01 mg/kg,低于国标液相色谱-质谱/质谱法,能满足酒中微量甜蜜素的检测要求。     

两种海藻物提取物的制备及脂肪酸GC-MS分析

制备三列凹顶藻和松节藻醇提物，采用气相色谱。质谱联用仪（GC／MS）对这两种海藻提取物脂肪酸进行了分析，各分离出17个和18个峰，鉴定了12种脂肪酸。用峰面积归一化法得出各类脂肪酸的相对百分含量。凹顶藻提取物中含饱和脂肪酸4种，总体百分含量为18．095％：单不饱和脂肪酸2种，总百分含量为3．910％；多不饱和脂肪酸种类较多，共有6种，总百分含量为7．437％。松节藻醇提物中4种饱和脂肪酸的含量为51．842％，2种单不饱和脂肪酸含量为13．124％，6种多不饱和脂肪酸含量为27．375％。     

Isolation and characterization of lecithin from squid (Todarodes pacificus) viscera deoiled by supercritical carbon dioxide extraction

Marine lecithin was isolated and characterized from squid (Todarodes pacificus) viscera residues deoiled by supercritical carbon dioxide (SC-CO(2)) extraction. SC-CO(2) extraction was carried out to extract the oil from squid viscera at different temperatures (35 to 45 °C) and pressures (15 to 25 MPa). The extraction yield was higher at highest temperature and pressure. The major phospholipids of squid viscera lecithin were quantified by high-performance liquid chromatography (HPLC). Phosphatidylcholine (PC; 80.5% ± 0.7%) and phosphatidylethanolamine (PE; 13.2% ± 0.2%) were the main phospholipids. Thin layer chromatography (TLC) was performed to purify the individual phospholipids. The fatty acid compositions of lecithin, PC and PE were analyzed by gas chromatography (GC). A significant amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were present in both phospholipids of PC and PE. Emulsions of lecithin in water were prepared through the use of a homogenizer. The oxidative stability of squid viscera lecithin was high in spite of its high concentration of long-chain polyunsaturated fatty acids. PRACTICAL APPLICATION: Squid viscera are discarded as a waste by fish processing industry. Since lecithin from squid viscera contains higher amounts of polyunsaturated fatty acids, it may have promising effect to use in food, pharmaceutical, and cosmetic industries.     

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。     

Effects of feeding tuna oil on the lipid composition of pig spermatozoa and in vitro characteristics of semen

The aim of the present study was to characterize the effects of feeding tuna oil on the lipid and fatty acid composition of boar spermatozoa and to relate changes in composition to boar semen characteristics. Ten boars were paired by age and allocated to one of two diets (five boars per diet). The diets, which were offered for 6 weeks, consisted of a basal diet that was either unsupplemented or supplemented with 30 g tuna oil kg(-1) diet. Adding tuna oil to the diet increased the ether extract concentration of the diets fed from 65 to 92 g kg(-1) dry matter and supplied 10.5 g long chain polyunsaturated (n-3) fatty acids per 100 g total fatty acids. There were no changes in semen fatty acid composition after 3 weeks of feeding tuna oil. However, after 5 and 6 weeks, the proportions (g per 100 g total fatty acids) of 22:6(n-3) in sperm phospholipid fatty acids were increased from 34.5 to 42.9 g by feeding tuna oil and 22:5(n-6) decreased from 29.8 to 17.9 g. No changes were observed in other sperm lipids or seminal plasma phospholipids as a result of the diets fed. Feeding tuna oil increased the proportion of spermatozoa with progressive motility and with a normal acrosome score and reduced the proportion of spermatozoa with abnormal morphologies.     

6种金枪鱼脂肪酸组成与含量差异性研究

目的比较捕捞于不同海域不同品种的6种金枪鱼的脂肪酸种类与含量。方法收集来源于大西洋、太平洋、印度洋的蓝鳍金枪鱼(Thunnus thynnus)、黄鳍金枪鱼(Thunnus albacores)、大目金枪鱼(Thunnus obesus)、马苏金枪鱼(Thunnus maccoyii)、长鳍金枪鱼(Thunnus alalunga)、鲣鱼(Katsuwonus pelamis)6种金枪鱼,采用三氟化硼甲醇甲酯化金枪鱼肌肉组织的脂肪酸,利用气相色谱法测定脂肪酸甲酯含量;再通过SPSS 24对各脂肪酸组成结果进行相关性分析。结果 6种金枪鱼肌肉脂肪酸种类与含量丰富,蓝鳍金枪鱼、大目金枪鱼等5种金枪鱼不饱和脂肪酸的含量明显高于鲣鱼,蓝鳍金枪鱼的不饱和脂肪酸种类最多且含量最高,捕捞于太平洋海域的大目金枪鱼、黄鳍金枪鱼的不饱和脂肪酸种类明显多于其他海域,人工饲养的马苏金枪鱼(太平洋)比野生马苏金枪鱼(印度洋)的不饱和脂肪酸种类丰富;6种金枪鱼主要组分间的相关性分析结果显示,C20:0与所测的9种脂肪酸呈负相关,C18:1与C18:2呈极显著正相关(r=0.900～1.000),C14:0与C17:1呈负相关,与其他8种脂肪酸呈正相关。结论 6种金枪鱼存在脂肪酸种类与含量的差异,不同海域与不同养殖方式下的金枪鱼脂肪酸的种类也存在差异。     

Effects of dietary corn oil and fish oil concentrate on lipid composition of calf tissues

Lipid composition of calf blood plasma, liver platelets, muscle, heart, and brain was measured, as affected by high dietary intake of linoleic acid from corn oil or of polyunsaturated fatty acids from fish oil concentrate. Plasma total lipids, phosphatidylcholine, and cholesteryl esters were reduced by corn oil and fish oil concentrate. Dietary fatty acid composition had no influence on percentage distribution of the major phospholipid components of liver, heart, muscle, and brain, but did alter the polyunsaturated fatty acid composition of major phospholipids in plasma, liver, platelets, muscle, and heart. In general, high linoleic acid intake increased linoleic acid and decreased oleic, arachidonic, and linolenic acids in tissue phospholipids, and fish oil concentrate high in eicosapentaenoic and docosahexaenoic acids increased phospholipid concentrations of these fatty acids. The fatty acid composition of brain phosphatidylcholine and phosphatidyl-ethanolamine was relatively resistant to dietary lipid alterations. The fatty acid changes in tissue phospholipids that resulted from dietary lipid alterations may have important implications in eicosanoid metabolism, and in the structure and function of cell membranes.     

Deodorisation of fish oil by nanofiltration membrane process: focus on volatile flavour compounds and fatty acids composition

This study focuses on the investigations for the deodorisation of tuna oil and squid oil through n‐hexane extraction and nanofiltration membrane process with a molecular weight cut‐off of 360 Da. The change of the odour and the fatty acids composition were analysed. Volatile components analysed by monolithic material sorptive extraction (MMSE) combined with gas chromatography–mass spectrometry (GC‐MS) suggested that nanofiltration membrane process could remove the majority of the volatile components, and the removal rate was approximate to 80%. The odour activity values (OAVs) of the volatile components in tuna oil and squid oil were significantly reduced from 125.23 to 17.47 and 129.76 to 10.06 (P < 0.05), respectively. On the other hand, the contents of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were increased as expected. These results indicated that the organic solvent nanofiltration technique had obvious effects on deodorisation of fish oil, and it would be a promising deodorisation method in fish oil refining industry.     

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.