Structures of the Ceramides from Porcine Palatal Stratum Corneum

Ceramides are the major type of lipid found in stratum corneum from the skin, gingiva and hard palate. The present study examined the ceramides of the stratum corneum from the hard palate. Six fractions of ceramides were isolated by preparative thin-layer chromatography. The least polar fraction contained an unusual acyl ceramide (EOS) consisting of long ω-hydroxy acids amide-linked to sphingosine with mostly saturated fatty acids ester-linked to the ω-hydroxyl group. The second and third fractions contained normal fatty acids amide-linked to sphingosine (NS) and phytosphingosine (NP), respectively. In each of these ceramides, the fatty acids consisted of a mixture of saturated and monoenoic species. The three most polar fractions all contain amide-linked α-hydroxy acids. The fourth fraction contained long α-hydroxy acids amide-linked to sphingosine (ASl), while the fifth fraction contained short α-hydroxy acids amide-linked to sphingosine (ASs). The most polar ceramide contained α-hydroxy acids amide-linked to phytosphingosine (AP). EOS, NS and NP differed from their epidermal counterparts in terms of the compositions of the normal fatty acids. ASl, ASs and AP from palatal stratum corneum were essentially identical to their epidermal counterparts. The differences between palatal and epidermal EOS, NS and NP contribute to the differences in permeability of palate compared to skin.     

The Triglyceride Composition of Mango (Mangifera indica) Kernel Fat

The triglyceride composition of the kernel fat of 9 different mango varieties has been determined. Stearic and oleic acids represent respectively from 32.7 to 44.0% and from 43.7 to 53.4% of the total fatty acids. The remaining fatty acids were palmitic (6.7-9.7%), linoleic (3.6-6.9%), arachidic (1.1-2.5%) and linolenic (0.3–1.0%) acids. The triglyceride components were determined by separating the triglycerides according their degree of unsaturation by means of thin-layer chromatography on silica gel impregnated with silver nitrate. The fatty acid composition of the different triglyceride fractions and of the fatty acids incorporated at the sn-2-position of each triglyceride fraction was determined. Moreover, the triglycerides were separated according to their carbon number by gas liquid chromatography using an open-tubular glass column, wall-coated with CP-Sil 5. The triglyceride compositions obtained by thin-layer chromatography on silica gel impregnated with silver nitrate were in agreement with the compositions predicted by the 1,3-random-2-random distribution hypothesis.     

Thin-layer chromatographic separation of dimethylphosphatidates derived from lecithins

The separation of lecithin derivatives based on their fatty acid substituents has been investigated. Several synthetic and natural lecithins were converted to their corresponding dimethylphosphatidates by hydrolysis with phospholipase D (phosphatidylcholine phospholipase D (phosphatidylcholine phosphatidohydrolase, EC 3.1.4.4) and methylation of the resulting phosphatidic acids with diazomethane. These dimethylphosphatidates were separated into fractions by reversed-phase thin-layer chromatography. Separations were dependent on the total number of methylene groups and double bonds in the two fatty acid chains. Fractionated dimethylphosphatidates were extracted from the plates and fatty acids were determined.     

Studies on Mirabilis jalapa (Four O'clock Plant) Seed Oil

Mirabilis jalapa of Nyctaginaceae plant family yields 4–5% of fatty oil. The oil is investigated for its glycerides and fatty acid composition by gas liquid chromatography. The fatty acids in the seed oil constitute C_16:0, 18.3%;C_18:1,55.3%;C_18:2,11.5%;C_18:3,14.9%. The triglyceride components were also determined by separating the triglycerides according to their degree of unsaturation by means of thin-layer chromatography on silica gel impregnated with silver nitrate. The fatty acid composition of the different triglyceride fractions was determined. Moreover, the triglycerides were separated according to their carbon number by gas liquid chromatography.     

The triglycerides of sable fish (anaplopoma fimbria). II. fatty acid distribution in triglyceride fractions as determined with pancreatic lipase

Sable fish muscle lipids were fractionated on a silicic acid column with mixtures of chloroform and methanol as eluting solvents. Three main peaks containing only triglycerides were isolated; 11 additional peaks contained phosphorous. Each of the 3 triglyceride peaks was separately fractionated into 300 fractions on silica gel columns impregnated with silver nitrate. Mixtures of petroleum ether and ethyl ether were the eluting solvents. About 25 distinct fractions were isolated from each column. The fractions were characterized for fatty acid content by gas chromatography of the methyl esters. The results showed that the fractionation did not depend upon the presence of single fatty acids but upon total unsaturation. Fatty acid distribution within each fraction was determined with the use of hog pancreatic lipase, followed by thin-layer chromatography and gas chromatography. Presented at the AOCS Meeting in Houston, Texas, 1965.     

Hydrierung von Lipiden als Hilfsmittel zur Identifizierung und Quantifizierung von Phosphatiden aus Membransystemen des Herzmuskels

Hydrogenation of Lipids for Identification and Quantification of Phosphatides from Pellicle Systems of Cardiac Muscle. It was the aim of our research to show that hydrogenation of lipids is an auxiliary technique in phospholipid analysis of cardiac membranes. This is of interest if a preliminary overview on lipid fractions containing highly unsaturated fatty acids is needed. The fatty acids and the diglycerides from phospholipids were hydrogenated according to the procedure described by Appelqvist (A simple and convenient procedure for the hydrogenation of lipids on the micro- and nanomole scale, J. Lipid Res. 13 (1972), 146) with platinum oxide as a catalyst. The lipids (fatty acid methyl esters or acetylated diglycerides) were taken to dryness in a test-tube under nitrogen and flushed with hydrogen. The catalyst, suspended in methanol was injected through a septum. For identification purposes thin-layer chromatography on silica gel and on silica gel impregnated with silver nitrate was combined with gas chromatography before and after hydrogenation. After hydrogenation the fatty acid profile is much simpler and the fatty acid methyl esters can easily be differentiated from dimethyl acetals, as the latter are more volatile. Diacylglycerides and alkenylacylglycerides were also separated by thin-layer chromatography in individual subclasses before they were analysed by gaschromatography. Hydrogenating the lipids makes it possible to circumvent in part difficulties which arise often with polyunsaturated fatty acids. As the chain length of C20 and C22 are mainly represented by C20:4 , the arachidonic acid and C22:6 the docosahexaenoic acid, both fatty acids can be assessed after hydrogenation. The fatty acid profile of phosphatidylcholine and phophatidylethanolamine of cardiac muscle from rat, guinea pig and pig was determined. Each sample was analysed before and after hydrogenation. The fatty acids with the same chain length were summed up and compared to the corresponding chain length after hydrogenation.     

Comparison of column chromatographic methods for the quantitative determination of mono- and digalactodiglycerides in fresh alfalfa (Medicago sativa)

Three column chromatographic procedures for separating and recovering the galactolipids in fresh alfalfa extracts were compared. Silicic acid chromatography yielded pure fractions by thin-layer chromatography, infrared absorption, and chemical analysis. The carbon-Celite column gave the highest yield of monogalactodiglyceride. Of the 1.2% total lipids of fresh alfalfa, approximately 12% was monogalactodiglyceride and 8% was digalactodiglyceride. Linolenic acid accounted for about 90% of the total fatty acids in these components.     

Composition of fatty acids and structure of triglycerides in medium and low erucic acid rapeseed

Total triglycerides in medium (MEAR) and low (LEAR) erucic acid cultivars of rapeseed were fractionated by argentation chromatography into twelve and ten fractions, respectively. Gas liquid chromatography of the fatty acids in the triglyceride fractions and their 2-monoglycerides was used to evaluate the structural characteristics of the individual fractions. Fractionation occurred on the basis of degree of unsaturation, molecular weight and positional characteristics. The most mobile fractions contained 34–50% of saturated fatty acids while the less mobile had 59–65% of polyunsaturated fatty acids. In the medium erucic acid oil, long chain fatty acids (C20–C22) were found in all fractions, but four fractions of low erucic acid oil were essentially free of long chain acids. Two of these fractions in the latter oil, which represented 44% of the total triglycerides, were glycerol trioleate and dioleoyllinoleoylglycerol. The majority of the 2-positions were occupied by unsaturated C18 fatty acids, generally in the order of linoleic ≥linolenic> oleic acids. The saturated and long chain fatty acids occurred predominantly in the 1-and 3-positions. The various fractions of medium and low erucic acid oils were similar in structural composition except that eicosenoic and erucic acids substituted for oleic acid in some external positions. Erucic acid did not appear to substitute directly for oleic acid in the 2-position.     

Chromatographic analysis of polyglycerols and their fatty acid esters

Polyglycerols and their fatty acid esters have been analyzed by gas-liquid chromatography (GLC) as trimethylsilyl ether derivatives. Linear diglycerols and triglycerols were isolated from commercial polyglycerols by vacuum distillation. Mono- and di-fatty acid esters were synthesized in the laboratory. Two isomers of diglycerol have been separated and identified. GLC analysis was carried out on columns packed with 3% JXR on Gas Chrom Q. Response factors for diglycerol and triglycerol relative to glycerol have been established. Commercial polyglycerol esters are shown to be mixtures of glycerol, free fatty acids, mono- and diglycerides, and mono- fatty acid esters of diglycerol and triglycerol. Separation of free polyglycerols and their esters is also demonstrated by thin-layer chromatography on Silica Gel-G containing 4.0% boric acid.     

Studies on the fatty acid composition of crayfish lipids

The fatty acid composition of carcass and exoskeleton lipids was determined for the freshwater crayfishOrconectes rusticus. Lipid fractions were isolated by column and thin-layer chromatography. Fatty acid methyl esters and alcohol acetates were then prepared and analyzed by gas-liquid chromatography. Peak identities were established from retention time data for methyl esters, hydrogenated methyl esters, and saturated, monoene, diene, and polyene methyl esters separated as acetoxy-mercuri-methoxy derivatives. Minor component acids were estimated from their relative compositions in these fractions. Presented at the symposium honoring J. B. Brown, AOCS meeting in Chicago, 1964.     

Fatty acid composition of free ceramides of kidney and cerebellum from a patient with Farber's disease

The fatty acid composition of ceramides has been determined in kidney and cerebellum of a patient with Farber's disease, which is characterized by ceramidase deficiency. Farber cerebellum and kidney contained a five- and ten-fold excess, respectively, of free ceramides. The nonhydroxy fatty acid patterns of the ceramides from Farber kidney and cerebellum showed considerable similarities to those from control tissues, whereas large amounts of ceramides containing hydroxy fatty acids are found in Farber's disease tissues.     

Lipids of hamster cheek pouch epithelium

The hamster cheek pouch is a much used but incompletely understood experimental model. In particular, the cheek pouch epithelial lipids, which are important for permeability barrier function as well as other aspects of epithelial biology, have not been completely characterized. In the present study, the complete lipid class composition has been determined by thin-layer chromatography in conjunction with photodensitometry. The major lipid classes were phospholipids, free sterols, and ceramides. Minor amounts of monohexosylceramides, sterol esters, fatty acids, and triglycerides were also present. Significant amounts of covalently bound ω-hydroxyceramide was also detected. Transmission electron micrographs reveal extensive, largely paired, lipid bilayers in the intercellular spaces of the stratum corneum.     

Lipid composition of ten edible seed species from North Vietnam

The lipid composition and oil content of ten edible seed species from North Vietnam(Cassia tora, Ipomoea aquatica, Raphanus sativus, Citrullus lanatus, Cucumis melo, Cucurbita pepo, Luffa cylindrica, Phaseolus vulgaris, Vigna aurea, Sesamum orientale) have been investigated. The contents of hydrocarbon, triacylglycerol, free fatty acid, sterol, di- and monoglycerol, and polar lipid fractions have been determined with a thin-layer chromatography (TLC)/flame-ionization detection analyzer. Molecular species of hydrogenated triacylglycerols and the fatty acid composition of total lipids also have been analyzed by capillary gas-liquid chromatography. The quantities of major phospholipid classes of four seed species(C. tora, I. aquatica, R. sativus, V. aurea) have been determined by two-dimensional TLC and the spectrophotometrical phosphorus analysis. The fatty acid compositions of nonpolar and polar lipid fractions of these four species also have been analyzed.     

Distribution of cholesteryl esters and other lipid in subcellular fractions of the adrenal gland of the pig

Total lipids from whole pig adrenal glands as well as from their mitochondria, microsomes, liposomes, and cell sap were extracted and fractionated first into neutral lipids and phospholipids. The highest percentage of neutral lipids was found in the cell sap, and the lowest in the microsomal fraction. Neutral lipids were subfractionated into cholesteryl esters, free cholesterol, triglycerides, and free fatty acids. Cholesteryl esters were distributed throughout the liposomes. Free fatty acids represented a substantial part of cell sap lipids, but were present also in the mitochondria, microsomes, and liposomes. Fatty acids of all fractions were analyzed by gas liquid chromatography. Free fatty acids and cholesteryl ester fatty acids from all cellular fractions were similar in composition and were characterized by considerable quantities of linoleic and arachidonic acid. Triglycerides were characterized by an increased percentage of palmitic and a low content of arachidonic acid. Phosphatidyl choline, phosphatidyl ethanolamine, diphosphatidyl glycerol, and sphingomyelin plus phosphatidyl inositol were isolated from the lipids by preparative thin layer chromatography, and their fatty acids analyzed by gas liquid chromatography. Phosphatidyl choline and phosphatidyl ethanolamine from mitochondria, microsomes, and cell sap were very similar in respect of their fatty acid composition. Sphingomyelin plus phosphatidyl inositol was characterized by a high content of C22:2omega6. Diphosphatidyl glycerol was present in mitochondria and in the cell sap.     

Use of acetyl chloride/methanol for assumed selective methylation of plasma nonesterified fatty acids results in significant methylation of esterified fatty acids

The albumin-bound nonesterified fatty acid pool in plasma, which represents a very small percentage of total plasma fatty acids, has previously been quantitated by a variety of methods. In the present study we determined that the nonesterified fatty acid concentrations in the plasma, quantitated by a popular method using acetyl chloride and methanol which is reported to be specific for methylation of nonesterified fatty acids in the presence of esterified fatty acids (i.e., without prior isolation of the plasma non-esterified fatty acids), were significantly overestimated due to cleavage and methylation of esterified fatty acids. Quantitation of the contaminating fatty acid from the esterified pool demonstrated that the amount of fatty acid cleaved from the esterified pool was enough to exceed the entire mass of nonesterified fatty acids. As an established method for comparison, we isolated nonesterified fatty acids from the plasma by thin-layer chromatography prior to methylation, using a number of simple precautions to limit oxidation. By performing all thin-layer chromatography steps in an atmosphere of nitrogen and by including fatty acid standards in the plasma with 0,1, 2 or 4 double bounds, we were able to accurately and reproducibly determine the concentration of nonesterified fatty acids in the plasma, including arachidonate. We demonstrated that no oxidation occurred in the thin-layer chromatographic isolation of homonesterified fatty acids and that the coefficients of variation for repeat measurements of the same sample were <11% using our reference method. Our data indicate that the use of acetyl chloride and methanol for assumed selective methylation of plasma nonesterified fatty acids results in significant methylation of esterified fatty acids.     

The isolation and characterization of lipid compounds from black soldier fly larvae

With the global demand for lipid compounds on the rise, concerns are growing about the environmental and economic impact of traditional lipid sources. This concern is exacerbated by the ever-increasing demand for plant-based lipids, which is contributing to unsustainable production practices and competition for land and food. Considering this challenge, this work aimed at exploring the potential of black soldier fly larvae (BSFL) as a low-cost and ecofriendly source of lipid classes. A fractionation scheme consisting of a mixture of polar and nonpolar solvents at different ratios was employed to isolate the lipid classes from BSFL oil using silica gel column chromatography, which is a conventional method of chromatography. The fraction's separation efficiency was validated using thin-layer chromatography and characterized with Fourier-transformed infrared spectroscopy and gas chromatography flame ionization (GC-FID). Triglycerides (30%) were found to be the most abundant component while cholesterols (0.6%) were the least abundant lipid fractions in the lipid mixture. GC-FID in the various lipid fractions analyzed, lauric acid exhibited the highest percentage among the triglycerides (16.64%), diglycerides (19.10%), monoglycerides (22.70%), and free fatty acids (27.65%) fractions. The fractionation scheme proposed achieves high efficiency in separating and recovering different lipid classes extracted from the BSFL.     

Identification of cyclopentenyl fatty acids by 1H and 13C nuclear magnetic resonance

Gorlic, chaulmoogric and hydnocarpic fatty acids, specific to the seed oil of the genus Hydnocarpus sp. (Flacourtiaceae), are determined only with difficulty by gas chromatography. These fatty acids were isolated in their methyl ester form by a combination of different chromatographic techniques (thin-layer chromatography/Ag⁺ and high-pressure liquid chromatography). The proton and carbon nuclear magnetic resonance analysis of these fatty acid methyl esters showed some characteristic signals of the cyclopentenyl ring. The presence of these signals in the proton and/or carbon nuclear magnetic resonance spectrum of an oil thus will allow us to confirm the presence of these cyclopentenyl fatty acids in lipids.     

Lipid changes in maturing oil-bearing plants

Seeds ofCrambe abyssinica C.D. 6619 and theBrassica napus varieties Golden and Zero-erucic were collected at different stages of maturity and the free lipid extracted with hexane. The lipid thus obtained was separated into lipid classes by silicic acid column chromatography. The lipid classes were further examined by thin-layer chromatography and the component fatty acids and sterols by gas-liquid chromatography. The relative amounts of the lipid classes in crambe and both rape varieties varied as the seed matured and a period of great change occurred about 10 days after fertilization. The greatest change was in triglycerides and phospholipids plus glycolipids. Free fatty acids, present in immature seeds, has almost disappeared at maturity. The lipid classes of crambe and both types of rape were in similar proportion at maturity. Differences in phospholipid and glycolipid composition were found between crambe and rape and between immature and mature rape. The fatty acid composition differred between lipid classes and changed with maturity. Changes in 18-carbon acids of Zero-erucic rape were concurrent with the development of erucic and eicosenoic acids in Golden rape. Contribution No. 36 of the Food Research Institute. Presented at the AOCS Meeting, Cincinnati, October 1965.     

Nachweis gesättigter cycloaliphatischer und aromatischer Fettsäuremethylester durch Dünnschicht- und Gas-Chromatographie

Detection of the Methyl Esters of Saturated Cycloaliphatic and Aromatic Fatty Acids with Thin-layer and Gas Chromatography Homologous series of methyl esters of saturated cycloaliphatic (CFA) and aromatic fatty acids (AFA), which were prepared by the alkali isomerisation of fish oil followed by hydrogenation and fractionation of the cyclised mixture as well as with the help of cyclising hydrogenation, were analysed by thin-layer and gas chromatography. Even 0.4% CFA and AFA, which do not form urea-adducts, can be detected in the fractions as a substance class in thin-layer chromatography. The equivalent carbon numbers (stearic acid = 18) of the principal isomers of CFA and AFA homologues (C₁₈ to C₂₂), which were obtained as a class in the thin-layer chromatographic separation, were determined by the gas chromatography.     

Chemoattraction ofBiomphalaria glabrata (Gastropoda: Planorbidae) to lipid standards and lipophilic factors in leaf lettuce and Tetramin

Chemoattraction of individualBiomphalaria glabrata snails for lipid standards and lipophilic fractions of leaf lettuce and Tetramin were studied in a Petri dish bioassay. Snails were more significantly attracted to a whole Tetramin lipophilic fraction than that of leaf lettuce. Thin-layer chromatography showed that major neutral lipid fractions in Tetramin were triacylglycerols, free fatty acids, and free sterols, and in leaf lettuce were free fatty acids and a mixed free sterol-chlorophyll fraction. Snails were significantly attracted to both the free fatty acid and free sterol fractions from Tetramin, but only to the free fatty acid fraction from leaf lettuce. Snails were significantly attracted to a mixed lipid standard containing equal amounts of phosphatidylcholine, cholesterol, oleic acid, triolein, and cholesteryl oleate. Of four individual neutral lipid standards tested, i.e., cholesterol, oleic acid, triolein, and cholesteryl oleate, snails were only attracted to cholesteryl oleate.