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 (λ_max at 410nm). 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 protein isolates were observed.     

Lipid content and fatty acid composition of green algae Scenedesmus obliquus grown in a constant cell density apparatus

The lipids of alga Scenedesmus obliquus grown under controlled conditions were separated and fractionated by column and thin-layer chromatography, and fatty acid composition of each lipid component was studied by gas-liquid chromatography (GLC). Total lipids were 11.17%, and neutral lipid, glycolipid and phospholipid fractions were 7.24%, 2.45% and 1.48% on a dry weight basis, respectively. The major neutral lipids were diglycerides, triglycerides, free sterols, hydrocarbons and sterol esters. The glycolipids were: monogalactosyl diglyceride, digalactosyl diglyceride, esterified sterol glycoside, and sterol glycoside. The phospholipids included: phosphatidyl choline, phosphatidyl glycerol and phosphatidyl ethanolamine. Fourteen fatty acids were identified in the four lipid fractions by GLC. The main fatty acids were C18:2, C16:0, C18:3(alpha), C18:1, C16:3, C16:1, and C16:4. Total unsaturated fatty acid and essential fatty acid compositions of the total algal lipids were 80% and 38%, respectively.     

Composition of wheat-flour lipids

Lipids were extracted from a single sample of wheat flour using three solvent systems: ethanol–diethyl ether–water (2:2:1 by vol.); chloroform–methanol (2:1 by vol.); and water-saturated n-butanol. Analysis of the extracts and of residual lipid in the extracted flour showed that water-saturated n-butanol was the most efficient solvent. Wheat-flour lipids were extracted with water-saturated n-butanol and separated by chromatographic procedures into individual components. The lipid classes which were isolated and studied were steryl ester, free sterol, 6-O-acyl steryl glucoside, steryl glucoside, triglyceride, diglyceride, monoglyceride, free fatty acid, monogalactosyl diglyceride, 6-O-acyl monogalactosyl diglyceride, digalactosyl diglyceride, digalactosyl monoglyceride, monoglycosyl ceramide, diglycosyl ceramide, N-acyl phosphatidyl ethanolamine, N-acyl lysophosphatidyl ethanolamine, phosphatidyl ethanolamine, lysophosphatidyl ethanolamine, phosphatidyl choline, lysophosphatidyl choline, phosphatidyl serine and phosphatidyl inositol. Monogalactosyl monoglyceride was also tentatively identified. The quantitative distributions of the lipid classes were determined. Monoglycosyl ceramide contained small amounts of normal fatty acids (12:0–24:0) and large amounts of 2-hydroxy fatty acids (principally 16:0 and 20:0), with similar amounts of dihydroxy long-chain bases (18:0 and 18:1) and trihydroxy long-chain bases (18:0, 18:1, 19:0, 19:1, 20:0, 22:0). The principal sterols were identified as β-sitosterol, campesterol, and C₂₈ and C₂₉ saturated sterols. The fatty acids in the sterol lipids were principally 16:0 (50–60%) and 18:2 (28–30%) with small amounts of 16:1, 18:0, 18:1 and 18:3. The fatty acids in all the glycerides were principally 18:2 (51–84%) with lesser amounts of 16:0, 18:0, 18:1 and 18:3.     

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.     

Phospholipids of marine origin. V. The crab--a comparative study of a marine species (Cyclograpsus punctatus) and a fresh water species (Potamon)

A comparison has been made between phospholipids extracted from a marine crab and from a fresh water crab. Marine crab (body and viscera) contained a phospholipid fraction which liberated 2-aminoethylphos-phonic acid upon hydrolysis, while this substance was absent from the fresh water crab. Marine crab phospholipids had a higher content of phosphatidyl choline (57%), a higher content of phosphatidyl serine (5%) but a lower content of phosphatidyl ethanolamine (22%) than phospholipids from the fresh water crab (respective values: 52, 2 and 27%). Marine crab phospholipids and non-phosphorylated lipids were richer in C20 plus C22 fatty acids but poorer in C18: 2 acid, than were the corresponding lipids from the fresh water crab.     

Changes in Fatty Acid Composition of Papaya Lipids (Carica papaya) During Ripening

Lipids from papayas (Carica papaya L.) Solo cultivar at various stages of ripeness were extracted and fractionated into neutral lipids (NL), glycolipids (GL), and phospholipids (FL). The major fatty acids of the three lipid classes at all levels of ripeness were C16:0, C18:1, C18:2, C18:3. NL increased in unsaturation with maturity as indicated in the decrease of the various saturated/unsaturated fatty acid ratios, GL decreased in the ratio of C16:0/C16:1 due to an increase in C16:1 fatty acid with increasing fruit maturity. The remaining fatty acids in GL remained relatively unchanged. The phospholipid fraction increased in unsaturation with maturity due to increases in C16:1 and C18:3 fatty acids.     

Phospholipids of marine origin. The lantern fish (Lampanyctodes hectoris)

Lantern fish of the species Lampanyctodes hectoris were shown to contain phospholipids (10 g kg^?1) and non-phosphorylated lipids (140 g kg^?1). The phospholipid fraction consisted of phosphatidyl choline (47% of total phospholipids), phosphatidyl ethanolamine (42%), phosphatidyl serine (3%), phosphatidyl inositol (1%), sphingomyelin (4%), lyso-phosphatidyl choline (1%) and cardiolipins (2%). Lantern fish (L hectoris) meals normally contain unacceptably high lipid contents (150 g kg^?1 and over); this characteristic was found not to be due to a high phospholipid level in the lantern fish. The major fatty acid of the phospholipids was C22:6n-3 (25% total fatty acids) followed by C16:0 (18%), C18:ln-9 (16%) and C20:5n-3 (8%). This distribution was different from that of the non-phosphorylated lipids where the major fatty acid was C16:0 (21%) followed by C18:ln-9 (19%), C20:5n-3 (11%), C20:l (7%) and C22:6n-3 (7%). The lantern fish press oil and residual meal lipids had fatty acid distributions similar to those of the non-phosphorylated lipids.     

Characteristics of lipids in buckwheat grain and isolated starch and their changes after hydrothermal processing

Studies were carried out on bond and free lipids of buckwheat grain and isolated starch. As regards free and bond lipids, fraction composition (percentages of neutral and polar fraction) and fatty acids were determined. It was found that the content of free lipids in buckwheat grain was twice higher than of bond lipids. On the other hand, in case of starch percentage of free lipids was twice lower than of bond lipids. As regards free and bond lipids of buckwheat grain and starch, main fraction was represented by the neutral one. Percentages of glycolipids and phospholipids amounted to only from 5 to 7%. Fatty acids were predominated by unsaturated acids C_18:2 and C_18:1. Fire and steam hydrothermal processing resulted in several quantitative changes, the most important ones being an increase of free lipid content in the grain and of bond lipid content in the starch, changes in the percentage of glycolipids and phospholipids, an increase of saturation of free lipid fatty acids in the grain, and of saturation of bond lipid fatty acids in the starch.     

LIPIDS OF CURED CENTENNIAL SWEET POTATOES

SUMMARY –Lipids isolated from cured Centennial sweet potatoes were identified and quantitated by a combination of column and thin layer chromatography. These lipids were shown to consist of 42.1% neutral lipids, 30.8% glycolipids and 27.1% phospholipids. Triglycerides and steryl esters were the major lipids of the neutral fraction. Among the phospholipids, phosphatidyl ethanolamine, phosphatidyl choline and phosphatidyl inositol were the most abundant. Galactolipids and the steryl glucosides were also present. The predominant fatty acids were stearic, palmitic, oleic, linoleic and linolenic.     

Fumgal lipids. I. Effect of different nitrogen sources on the chemical composition

The effect of three nitrogen sources on the chemical composition of seven fungi, namely: Aspergillus niger, A. luchuensis, Penicillium crustosum, Alternaria tenuis, Rhizoctonia solani, Mucor sp. and Pythium irregulare has been studied. The various fungi showed a great variation with respect to lipid percentage and total lipid content. Lipid content varied from 3.2 to 41.5%. Non-polar lipids were comprised of monoglycerides, diglycerides, free sterols, free fatty acids and triglycerides. The quantitative make-up of the non-polar lipid varied with different nitrogen sources. Palmitic, stearic, oleic and linoleic acids were the major fatty acids while lauric, myristic, palmitoleic, linoleic and arachidic were the minor ones. The fatty acid composition was dramatically changed by changing the nitrogen source. Since different fungi responded differently to changes in nitrogen source, no generalisation could be made. Two-dimensional thin-layer chromatography of the polar lipid fraction of these fungi revealed the presence of a maximum of fifteen spots. Phosphatidyl choline, phosphatidyl ethanolamine and phosphatidyl inositol were the major spots while lysophosphatidyl choline, lysophosphatidyl ethanolamine and phosphatidyl glycerol were present in smaller quantities. In comparison to phospholipids, glycolipids (except sterol glycoside) were present in relatively lower concentration. Pythium irregulare was very characteristic in having no glycolipids at all.     

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.     

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。     

Seasonal changes in phospholipids of mussel (Mytilus edulis Linne)

The phospholipids of mussels (Mytilus edulis Linne) from the coast of Qingdao were extracted, fractionated and analysed over a 12 month period. The contents of total lipids, neutral lipids, polar lipids and phospholipids were measured. The composition of phospholipids was determined by high‐performance liquid chromatography, and their fatty acid composition was analysed by gas chromatography. The phospholipid content ranged from 3.6 to 6.4 g kg^?1 soft tissue. PE (phosphatidyl ethanolamine) and PC (phosphatidyl choline) were the major constituents. C16:0, C20:5 and C22:6 were the major fatty acids. C20:5 (5.25–23.10%) and C22:6 (6.05–20.42%) varied regularly with the seasonal factors. Their total amounts were high from January to June, which would be an optimal time for the utilisation of the phospholipids of mussels. © 2002 Society of Chemical Industry     

Selective extraction and quantitative analysis of non-starch and starch lipids from wheat flour.

Wheat flour non-starch lipids (lipids not bound to starch) were quantitatively extracted with water-saturated n-butanol (WSB), benzene-ethanol-water (10:10:1, by vol.) or ethanol-diethyl ether-water (2:2:1, by vol.) in 10min at 20 °C. Starch lipids (lipids bound to starch) were subsequently extracted with WSB at 90–100 °C. Carotenoid pigments were quantitatively extracted with the non-starch lipids. There was no significant hydrolysis of esterified fatty acids and no detectable autoxidation of unsaturated acids in the hot solvent extracts. Non-starch and starch lipids from a high grade spring wheat flour and three grades of winter wheat flour were separated by thin-layer chromatography and quantified as fatty acid methyl esters (FAME) by gas-liquid chromatography (g.l.c.) using heptadecanoic acid (or its methyl ester) as internal standard. Total flour and starch lipids after acid hydrolysis were also converted to FAME for quantitation by g.l.c. Non-starch lipids consisted of 59–63% non-polar (neutral) lipids, 22–27% polar glycolipids and 13–16% phospholipids. Steryl esters, triglycerides, and all the diacyl galactosylglycerides and phosphoglycerides were present only in non-starch lipids. Starch lipids consisted of 6–9% non-polar (neutral) lipids (mostly free fatty acids), 3–5 % polar glycolipids and 86–91 % phospholipids (mostly lysophosphatidyl choline). Starch lipids were almost exclusively monoacyl lipids. Factors are given for converting weight of FAME into weight of original lipid for all individual lipid classes in wheat which contain O-acyl groups. Factors for total lipids are: total starch lipids = FAME × 1.70, total non-starch lipids = FAME × 1.20, and total flour (non-starch + starch) lipids = FAME × 1.32. Similar factors could be used to convert weight of lipids obtained by conventional acid hydrolysis methods into weight of unhydrolysed lipids. Phospholipid contents are given by: total starch phospholipids = P × 16.51, total non-starch phospholipids = P × 23.90, total flour phospholipids = P × 17.91.     

A Research Note Lipid Composition of Cumin (Cuminum cyminum L.) Seeds

Total lipids extracted from Cumin (Cuminum cyminum L., Umbelliferae) amounted to 14.5% of the dry seeds. The total lipids consisted of 84.8% neutral lipids, 10.1% glycolipids, and 5.1% phospholipids. Neutral lipids consisted mostly of triacylglycerols (89.4%) and small amounts of diacylglycerols, free fatty acids, sterols, sterolesters, and hydrocarbons. At least five glycolipids and five phospholipids were identified. Acylmonogalactosyldiacylglycerol and acylatedsterylglucoside were the major glycolipids, while monogalactosyldiacylglycerol, monogalactosylmonoaclycerol, and digalactosyldiacylglycerol were present in small quantities. Phosphatidylethanolamine and phosphatidylcholine were the major phospholipids, while phosphatidylinositol, lysophosphatidylethanolamine, and phosphatidylglycerol were present in small amounts. The fatty acid composition of these different neutral lipids, glycolipids, and phospholipids was determined     

Changes in milk fat phospholipids during lactation

Changes in lipid composition were studied in milk obtained on postpartum d 3 (colostrum), 7, 42, and 180 from 12 Holstein cows. Triglycerides, 96 to 97% of total lipids, were relatively constant during lactation. Phospholipids and cholesterol declined with advancing lactation. Concentrations of the fatty acids synthesized within the mammary gland, C10:0 to C16:0, increased about 50% from 7 to 42 d of lactation. During this period, compensatory decreases were observed in C18:1. The phospholipids were separated into five major classes: sphingomyelin, phosphatidyl choline, serine, inositol, and ethanolamine for fatty acid analysis. The changes that occurred in milk total fatty acids were reflected in phosphatidyl phospholipid fatty acid composition: an increase in medium-chain fatty acids and a decrease in polyunsaturated fatty acids of 18, 20, and 22 carbon atom chain length as lactation progressed. These changes are consistent with the theory that milk phospholipids are synthesized de novo entirely in the mammary gland.     

Lipids in cereals. 1. Pennisetum typhoideum

Two improved strains of Pennisetum typhoideum (‘bajra’) were found to have a free lipid content of about 5.0% and bound lipid content of about 0.5%. In the non-polar fraction, sterol esters and hydrocarbons, triglycerides, free fatty acids, free sterols and partial glycerides were present with triglycerides as the principal constituents. Polar lipids were separated by two-dimensional thin-layer chromatography and lecithin was found to be the major component. Sterol-containing glycolipids (sterol glycosides and esterified sterol glycosides) were present in appreciable amounts. Phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl inositol, lysophosphatidyl ethanolamine, lysolecithin, phosphatidic acid, poly-glycerophosphatide, mono- and di-galactosyl glycerides and cerebrosides have also been tentatively identified.     

Composition of the lipids of lupin seed (Lupinus angustifolius L. var. “uniwhite”)

The total lipids (8.6%) extracted from whole lupin seeds (Lupinus angustifolius L. var. “Uniwhite”) were found to be comprised of triglycerides (71.1%), phospho-lipids (14.9%), free sterols (5.2%), glycolipids (3.5%), sterol and wax esters (0.5%), free alcohols (0.4%), hydrocarbons (0.4%) and unidentified waxy material (4.0%). The main fatty acids in the total lipid extract were linoleic (48.3%), oleic (31.2%), palmitic (7.6%) and linolenic (5.4%). Erucic acid was not present, nor were cyclopropenoid acids. Seed coatings constituted 23.9% of the weight of the whole seeds and contained 1.5% lipids, the main classes of which were triglycerides (38.4%), free sterols (28.0%), phospholipids (9.7%), glycolipids (9.1%) and free alcohols (3.7%). The seed coatings and kernels contained the same fatty acid constituents, but the proportions of oleic, linoleic and linolenic acids were markedly different. β-Carotene was present, although at low concentration. When evaluated on its lipid composition, “Uniwhite” lupin seed appears suitable as a supplement for pig, poultry and stock feeding.     

Studies on Compounds and Individual Lipids of Wheat Germ

SUMMARY: A combination of chemical and chromatographic procedures demonstrated that 97% of total wheat germ lipids are nonpolar, while 3% are polar (glycolipids, phosphoipids and proteolipids). The fatty acid composition, in polar and nonpolar lipids, was determined: the former contain a higher percentage of octa-decatrienoic acid than the latter. Nonpolar lipids are tri, mono- and diglycerides and sterol esters. In addition to lecithin, phosphatidyl ethanolamine, phosphatidyl serine and their lyso derivatives, lipids contain unknown compounds tentatively identified as glycolipids or phytoglycolipids. Moreover a high content of compounds identified as phosphatidic acid or polyglycerophosphate derivatives has been observed.     

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。