Lipid Composition of Garlic

Neutral, glyco- and phospholipids of garlic were resolved into their component fractions by thin layer chromatography. Neutral lipids contained considerable quantities of monoglycerides (18.5%), diglycerides (14.2%), sterols (16.3%) and triglycerides (41.5%) respectively. The phospholipid fraction was rich in phosphatidyl choline (23.5%), phosphatidyl ethanolamine (17.9%), lysophosphatidyl choline (11.8%) and lysophosphatidyl ethanolamine (8.2%). Digalactosyl diglyceride (10.1%), sterol glycoside (15.6%), cerebrosides (8.1%), acylsterol glycoside (38.6%) and monogalactosyl diglyceride (22.5%) were the major components of the glycolipids of garlic. Lauric, myristic, palmitic and linoleic acids constituted the major fatty acids of monoglycerides, diglycerides and free fatty acid fractions whereas palmitic, linoleic and linolenic acids were the major fatty acids of triglycerides. Palmitic and linoleic acids were the major fatty acids of garlic phospholipids. Except the acylsterol glycoside fraction glycolipids were rich in lauric, palmitic, linoleic and linolenic acids; palmitic acid was the only major fatty acid of acylsterol glycosides.     

The major component phospholipids of rapeseed gum

The phospholipids from a commercial rapeseed gum have been fractionated on DEAE-cellulose and silicic acid columns. The molar percentages of the major components were phosphatidyl choline (22), phosphatidyl inositol (18) and phosphatidyl ethanolamine (15). Other acidic phospholipids (16) were also observed but were not further investigated. The fatty acids from the phospholipid fractions showed little variation in composition. The chief components were palmitic, oleic and linoleic acids. Issued as NRC No. 8947. National Research Council of Canada Postdoctorate Fellow 1964–65.     

Phospholipids of the pulmonate land snailCepaea nemoralis (L.)

The phospholipids of the snailCepaea nemoralis, comprising the major lipid fraction (65%) in this terrestrial pulmonate, were investigated by thin-layer and column chromatography. Detailed gas chromatographic analyses of liberated fatty acid fractions and amino acid analyses of the water soluble moieties of isolated phospholipid classes were carried out. Phosphatidyl choline (47%) and phosphatidyl ethanolamine (21%) were found to be the predominant phospholipid classes, while phosphatidyl serine (8%), phosphatidyl inositol (6%), diphosphatidyl glycerol (3%), ceramide amino-ethylphosphonate (7%), lysophosphatidyl choline (1%), and phosphatidic acid (1%) were present in lesser amounts. In the phosphatidyl ethanolamine and phosphatidyl serine fractions, minor quantities of plasmalogen analogues were detected. Fatty acid profiles of the various phospholipid classes appeared to be strikingly diverse, e.g. a characteristic component, such as linoleic (18∶2ω6) acid, ranging from 3–54%. In vivo radioisotope studies using 1-¹⁴C-acetate demonstrated the high biosynthetic rate of all phospholipid classes and their respective fatty acid fractions. Results are discussed in relation to data on the phospholipids from other invertebrate species.     

Phospholipid composition of the boll weevil,Anthonomus grandis boheman

When phospholipids of newly-emerged adults of the boll weevil,Anthonomus grandis Boheman, were studied in detail, phosphatidyl choline and phosphatidyl ethanolamine were found to be the major phospholipids; sphingomyelin and cardiolipin were present in smaller amounts, and four other minor components were identified. Fatty acid analyses performed on the intact phospholipids and on the enzyme degradation products of phosphatidyl choline and phosphatidyl ethanolamine demonstrated that oleic and linoleic acids were the major fatty acids present in the glycerophosphatides; the sphingomyelin contained fatty acids in the range of 20–22 carbons.     

Phospholipid Composition of <Emphasis Type="Italic">Jatropha curcus</Emphasis> Seed Lipids

Jatropha curcus L. oil has emerged as one of the most important raw materials for biodiesel production. However, no detailed study has been reported on characterizing the lipid constituents of jatropha oil. The present study revealed that the total oil content of jatropha seeds was 32% with a composition of 97.6% neutral lipids, 0.95% glycolipids and 1.45% phospholipids. The fatty acid composition of total lipids, neutral lipids, phospholipids and glycolipids was also determined and found to contain oleic acid (18:1) and linoleic acids (18:2) as major fatty acids. The phospholipids fraction was further characterized and quantified and found to contain phosphatidyl choline (PC) 60.5%, phosphatidyl inositol (PI) 24% and phosphatidyl ethanolamine (PE) 15.5%. The fatty acid composition and the positional distribution of the fatty acids of individual phospholipids were also reported.     

The lipid components of white potato tubers (Solanum tuberosum)

Four Canadian varieties of potatoes were examined for their lipid composition. Lipids, extracted with chloroformmethanol, were shown by TLC and column chromatography to consist of 16.5% neutral lipids, 45.5% phospholipids and 38.1% glycolipids. Among the phospholipids and glycolipids, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol, the galactolipids and the sterol glucosides were the major lipids. The predominant fatty acids were palmitic (19.5%), linoleic (44.8%) and linolenic (30.4%, in Kennebec). Analyses of the fatty acids of stored potatoes showed a marked decrease in linoleic acid and an increase in linolenic acid, in the Irish Cobbler and Sebago potatoes. β-sitosterol comprised 85.0% of total sterols. Nearly half of the carotenoids was lutein (xanthophyll), the others being α-carotene, β-carotene, an unidentified pigment and lutein epoxide. Contribution No. 101 of the Food Research Institute, Canada Department of Agriculture.     

Composition and structure of phospholipids in chicken muscle tissues

Lipids extracted from breast muscle and thigh muscle of one-year old chickens on a standard MSU-Z-4 diet have been fractionated by silicic acid column chromatography into nonphospholipids, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl choline (lecithin), and sphingomyelins. Phospholipid fractions were identified by thin-layer chromatography and the quantity of each determined by gravimetric analysis, analysis of the phosphorus content, and infrared spectra. The phospholipid content of thigh muscle (dark meat) lipids was higher than that in the breast muscle (white meat). Phosphatidyl choline and phosphatidyl ethanolamine were found in relatively greater amts than phosphatidyl serine and sphingomyelins. Enzymatic hydrolysis followed by gas-liquid chromatographic analysis of the fatty acids liberated and those in the lysocompounds was used to establish the positional specificity of the fatty acids in the phosphoglycerides. The polyunsaturated fatty acids are located primarily at the β-position and the saturated fatty acids at the α′-position. The qualitative and quantiative determination of the plasmalogens was also accomplished. Michigan Agriculture Experiment Station Journal Article No. 3527. Presented at the AOCS Meeting in Chicago, October, 1964.     

Phospholipids of human serum

Phospholipids extracted from normal human serum were fractionated into lecithin, lysolecithin, sphingomyelin, phosphatidyl ethanolamine, lysophosphatidyl ethanolamine, phosphatidyl serine, and phosphatidyl inositol. Identification of each was established by thin-layer chromatography and infrared spectrophotometry. The content of plasmalogen was determined in both lecithin and phosphatidyl ethanolamine fractions. The composition of fatty acids and fatty aldehydes in isolated phospholipids is presented. The degree of unsaturation as reflected in the average content of double bonds per molecule of the fatty acids in phospholipids was: lecithin 1.2, choline plasmalogen 2.1, lysolecithin 0.6, sphingomyelin 0.2, phosphatidyl ethanolamine 2.8, lysophosphatidyl ethanolamine 1.0, phosphatidyl serine 1.0, and phosphatidyl inositol 1.8. Both chlline and ethanolamine plasmalogen aldehydes were predominantly saturated. Molecular weight of each phospholipid was calculated from determined fatty acid and fatty aldehyde compositions; the phosphorus factor for each phospholipid was computed. On a weight percent basis, lecithin, sphingomyelin, and lysolecithin accounted for 95% of the total phospholipids. The ethanolamine-containing phospholipids accounted for 2.5%, and the remainder was divided among phosphatidyl inositol, choline plasmalogen and phosphatidyl serine. Presented in part at the AOCS Meeting, Houston, April, 1965. Dept. of Health, Education and Welfare, USPHS.     

Methodology for the separation o plant lipids and application to spinach leaf and chloroplast lamellae

Procedures for separation of complex plant lipids and results obtained are reviewed. Procedures based on DEAE cellulose and silicic acid chromatography, which may be preceded by countercurrent distribution, are presented for separation of the individual glyceroland sphingolipid classes of spinach leaf and chloroplast lamellae. These procedures appear to be generally applicable to photosynthetic tissue of plants and algae. The separation and infrared spectra of mono-and digalactosyl diglycerides, lecithin, phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl inositol, plant sulfolipid, cerebroside, and sterol glycosides from spinach are recorded. Chloroplast lamellae lipids are in the molar ratio monogalactosyl diglyceride (14.0), digalactosyl diglyceride (8.0), phosphatidyl glycerol (5.5), sulfolipid (3.9), lecithin (2.0), phosphatidyl inositol (1.0). Phosphatidyl ethanolamine, cerebrosides, and sterol glycosides were not detected in chloroplast lamellae. Fatty acid composition of individual lamellae lipids have been determined: The galactosyl lipids contain more than 90% trienoic acids.Trans-3- hexadecenoic acid is restricted almost exclusively to phosphatidyl glycerol. In this report techniques which have been applied to the isolation of plant glycero- and sphingolipids are reviewed and a new scheme presented for the separation of several of the plant lipid classes. Results obtained with spinach leaf and its photosynthetic apparatus are presented and discussed. Paper III in the series Plant and Chloroplast Lipids. Presented at the 15th Annual Summer Program, “Symposium on Quantitative Methodology in Lipid Research”, Aug. 3–7, 1964. Literature is reviewed to July 1964.     

The effect of diet on the phospholipid composition of the red blood cells of man

Short term (16 day) controlled fat (formula type diet) feeding to 10 healthy adult males led to no detectable change in the total amt or the relative proportions of the individual phospholipids of the red blood cells, although limited changes did occur in the fatty acids of certain of the phospholipids. The total phospholipid content of the red blood cells was 315±10 mg/100 ml (average of 20 samples). Lecithin accounted for 34% of the total, with sphingomyelin, phosphatidyl ethanolamine and phosphatidyl serine representing 25, 25 and 16%, respectively. Approx 36% of the phosphatidyl ethanolamine, 4% of the phosphatidyl serine and 6% of the lecithin was present in the plasmalogen form. Each phospholipid class was found to have a distinctive fatty acid spectrum. The M ratio of saturated to unsaturated fatty acids in all three phosphoglycerides was nearly 1:1. Behenic, lignoceric and nervonic acids made up almost half of the sphingomyelin fatty acids, and the M ratio of saturated to unsaturated fatty acids in this lipid was 3:1. When compared with red cells from subjects consuming a diet with a high butter fat content, red cells from subjects on a diet rich in corn oil were found to contain higher levels of linoleic acid in the lecithin and phosphatidyl serine fractions, and lower levels of oleic acid in the lecithin fraction. No changes were observed in the fatty acids of the phosphatidyl ethanolamine and sphingomyelin fractions. It is probable that these alterations represent the result of highly specific exchanges with plasma fatty acids, and it is suggested that three levels of specificity are involved: class of phospholipid, type of fatty acid, and specific fatty acid.     

Lipid composition of beef and human pituitary glands

The lipid composition of beef and human pituitary was determined by chromatographic and spectrophotometric methods. Beef pituitary lipid contained about 25% nonpolar lipids and 75% phospholipids whereas nonpolar lipids made up approximately 60% of the total in human pituitaries. The main nonpolar (i.e., low polarity) lipids in human pituitary were triglycerides, cholesterol, free fatty acids and an unidentified component in the triglyceride fraction. Cholesterol was the major nonpolar lipid component in freshly collected beef anterior and posterior pituitary, but the amount of free fatty acids appeared to increase during storage. Preliminary investigation of the unknown nonpolar lipid in human pituitaries suggested that it was an unsaturated hydroxy compound with no carbonyl functions. Thin layer chromatography indicated that it was also present in smaller amounts in freshly collected beef pituitaries. The main phospholipids of beef anterior, posterior and human pituitary were phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl inositol, phosphatidyl serine and sphingomyelin. The fatty acid composition of total nonpolar lipids, free fatty acids, total phospholipids, phosphatidyl ethanolamine and phosphatidyl choline of beef anterior and posterior pituitary was determined by gas liquid chromatography. Mixtures of saturated and unsaturated fatty acids ranging from C₁₂ to C₂₂ were present; the main fatty acids were palmitic, stearic, oleic, linoleic and arachidonic.     

Lipids of cultured hepatoma cells: VI. Glycerolipid and monoenoic fatty acid biosynthesis in minimal deviation hepatoma 7288C

1-14C-Acetic, 1-14C-palmitic, or 1-14C-stearic acid was incubated with minimal deviation hepatoma 7288C cells grown in culture to assess: de novo fatty acid synthesis, oxidation, desaturation, and elongation of saturated fatty acids, as well as the ability of media fatty acids to serve as precursors of cellular glycerolipids. Distribution of radioactivity in the individual lipid classes and the various fatty acids of triglyceride, phosphatidyl choline, and phosphatidyl ethanolamine was determined. The radioactivity among the monoenoic acid isomers derived from triglyceride, phosphatidyl choline, and phosphatidyl ethanolamine was analyzed by reductive ozonolysis. Only small amounts of the labeled substrates were oxidized to carbon dioxide. Except for labeled stearic acid, which also was incorporated heavily into phosphatidyl inositol and phosphatidyl serine, most radioactivity was recovered in triglyceride, phosphatidyl choline, and phosphatidyl ethanolamine. Synthesis of cholesterol and long chain fatty acids from labeled acetic acid demonstrated that these cells can perform de novo synthesis of fatty acids and cholesterol. Both labeled palmitic and stearic acids were desaturated to the corresponding delta9 monoenes, and palmitic and palmitoleic acids were elongated. The nexadecenoic acid fraction isolated from triglyceride, phosphatidyl choline, and phosphatidyl ethanolamine, when acetic or palmitic acid was the labeled substrate, showed that greater than 70 percent of the labeled acids were the delta9 isomer. Radioactivity of the octadecenoic acid fraction derived from labeled acetic or palmitic acids was nearly equally divided between the delta9 isomer, oleic acid, and the delta11 isomer, vaccenic acid. Desaturation of labeled stearic acid produced only oleic acid. These data demonstrate that the biosynthesis of vaccenic acid in these cultured neoplastic cells proceeds via the elongation of palmitoleic acid. The relatively high level of vaccenic acid synthesis in these cells suggests that the reported elevation of "oleic acid" in many neoplasms may result from increased concentration of vaccenic acid.     

Phosphatides isolated from seeds of commercial and experimental safflower varieties

Studies on phosphatides from several safflower varieties show the following five major results. The total phosphatide contents of the various safflower seeds are quite similar (0.48% for a commercial and 0.58% for a brown-striped variety). The same three major and one minor phosphatides were present in all varieties: phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI) and phosphatidyl serine (PS). The amounts of these lipids present in the crude phosphatide mixture were quite similar in all varieties tested (}36% for PC, }15% for PE, }23% for PI, and less than 2% for PS). The fatty acid composition of the phosphatides of UC-1 high oleic safflower is very different from that of the other varieties, but it reflects the composition of the corresponding oil triglycerides as far as the major acid is concerned. All other safflower seed phosphatides investigated have linoleic acid as the major fatty acid constituent. A simple and very sensitive color test has been found which can differentiate phosphatides of the high linoleic from the high oleic type. W. Utiliz. Res. Dev. Div., ARS, USDA.     

Molecular species of phosphatidyl ethanolamine from egg yolk

Phosphatidyl ethanolamine was isolated from total egg yolk lipids by preparative thin layer chromatography (TLC). The purified phosphatide contained 3% of the alkoxy derivative. It was degraded to diglycerides in the presence of purified sphingomyelin by phospholipase C fromClostridium welchii. The diglycerides were acetylated and resolved on the basis of unsaturation by argentation TLC. The fatty acid composition of the original phosphatidyl ethanolamine and the derived acetates was determined by gas chromatography, as was the molecular weight distribution of the diglyceride acetates. The placement of the fatty acids in the parent phosphatide was deduced by hydrolysis with phospholipase A fromCrotalus atrox, and in the acetates with pancreatic lipase. Some 33 major species of phosphatidyl ethanolamine were identified and compared to those for egg yolk lecithins. Presented in part at the Canadian Federation of Biological Societies Meeting, Kingston, June 1968.     

Liver lipids during development

The fatty acid composition of the major liver microsomal phospholipids has been studied during pre- and postnatal development of the rabbit. The fatty acid composition of the total lipids, phosphatidyl choline, and phosphatidyl ethanolamine from animals −6, −3, 0, +3, +6, +9, +16, and +112 days of age was determined. Fatty acid composition is similar in phosphatidyl choline and phosphatidyl ethanolamine for oleic acid at +3, +6, +9, and +16 day old animals; palmitoleic acid at +9 day old animals and linoleic acid at −6, −3, and 0 day old animals. Palmitoleic acid demonstrated a uniform decrease during early development in the total lipids and in both phosphatidyl choline and phosphatidyl ethanolamine; however, in the 112 day animal, the amount was just slightly lower than that observed for the earliest prenatal animal studied. Oleic acid decreased considerably during early postnatal development in the total lipids, phosphatidyl choline and phosphatidyl ethanolamine, but an increase in the 112 day animal was observed. Linoleic acid fluctuated considerably throughout postnatal development in the total lipids as well as in the two major phosphatides. Lecithin biosynthesis has been studied by two pathways during development of rabbit liver from −6 days to +110 days. The two pathways of lecithin biosynthesis were evaluated by assaying the activities of the liver enzymes choline phosphotransferase and phosphatidylmethyltransferase at different time intervals during development. The greater enzymatic activity was observed in the cholinephosphotransferase during development.     

Lipid and fatty acid composition of testes of quaking mice

Testes of quaking mice (sterile mutants) and of controls were analyzed for major lipid classes and fatty acid composition. Of the main lipid classes, only cholesterol esters differed significantly in concentration between the two groups (1.01 for quakers vs 0.69 mg/g wet wt of tissue for controls). The concentration of triglycerides was 4.5–5.0, that of total phosphatides 18–19, and that of free cholesterol 1.9–2.0 mg/g for mutants and controls. The concentrations of phosphatidyl ethanolamine and of sphingomyelin were both lower in quaking than in normal mice, but only the change in the former was statistically significant. Phosphatidyl choline was the major phosphatide (43–45% of total phosphatides) followed by phosphatidyl ethanolamine (24–26%) and sphingomyelin, phosphatidyl serine, and phosphatidyl inositol (all ca. 7% of total phosphatides). Minor differences between the mutants and controls were observed in concentrations of fatty acids of major lipid classes. The mutants, sterile because of faulty spermatid differentiation, had normal quantities of 22∶6 w3 and 22∶5 w6. These data are consistent with the hypothesis that the 22-carbon polyenes are associated with the formation of spermatids, rather than with their final differentiation into spermatozoa.     

Lipid Studies of Pimpinella acuminata

The seed oil of Pimpinella acuminata species of the family Umbelli-ferae was extracted with chloroform : methanol (2:1) for the separation of different classes of lipids as hydrocarbons (1.7%), sterol esters (traces), triglycerides (74.1%), free fatty acids (6.6%), 1,3-diglycerides (1.6%), 1,2-diglycerides (1.6%), sterols (1.8%), mono-glycerides (2.0%), phosphatidyl ethanolamine (0.8%), phosphatidyl choline (1.2 %), lyso phosphatidyl ethanolamine (0.4%), phosphatidyl inositol (1.2%) and unknown (7.0%). The fatty acids as determined by application of gas liquid chromatography is C₁₀-C₂₂, whereas C_16:0, C_18:0-C_18:2 and C_18:2 are predominantly present in all polar and non-polar lipid classes.     

Phospholipid oxidation in emulsions

The autoxidation of purified phosphatidyl ethanolamine (PE) and phosphatidyl choline (PC), extracted from egg and soybean lipids, was followed by oxygen uptake measurements in emulsified systems. All emulsified phospholipid fractions had comparable activation energies. Measurement by various physico-chemical tests was made of specific changes in the phospholipid molecule during autoxidation. PE oxidized more rapidly and absorbed more oxygen than PC. Higher 2-thiobarbituric acid test and diene and triene conjugation absorbance values were observed for PE than for PC. Of the two major polyunsaturated fatty acids in egg phospholipids, arachidonic acid disappeared at a more rapid rate during oxidation while the concentration of linoleic acid decreased to a level that was relatively constant. Although typical unsaturated fatty acid oxidation appeared to occur in all phospholipid fractions, oxidation in aqueous emulsions was only partly a function of fatty acid composition. The nitrogen moieties, ethanolamine and choline influenced the induction period for the oxidation of PE and PC respectively. Michigan Agriculture Experiment Station Journal Article No. 4420. Presented in part at the AOCS Meeting, Chicago, October 1967.     

Lipids ofKlebsiella pneumoniae: The presence of phosphatidyl choline in succinate-grown cells

The carbon and energy source for aerobically grown cultures ofKlebsiella penumoniae profoundly influenced the total lipid content and phosphatide composition. Glucose-grown cells contained 13% lipid, 56% of which was phospholipids. Succinate-grow cells contained 8% lipid, 66% of which was phospholipids. The predominant phosphatides of glucose-grown cells were phosphatidyl ethanolamine, 82%; phosphatidyl glycerol, 4.5%; phosphatidic acid, 5%; cardiolipin, 6.5%; phosphatidyl serine; and trace amounts of unidentified phosphatides. Phosphatides of succinate-grown cells were phosphatidyl ethanolamine, 38%; diphosphatidyl glycerol, 14%; phosphatidyl glycerol, 13%; phosphatidyl choline, 14.5%; phosphatidyl serine, 6%; phosphatidic acid, 4%; and 10% unknown lipids. No trace of phosphatidyl choline was found in glucose-grown cells. Paper 75-11-170 of the Kentucky Agricultural Experiment Station.