The effect of lyophilization on the solvent extraction of lipid classes,fatty acids and sterols from the oysterCrassostrea gigas

The lipid class compositions of adult Pacific oysters [Crassostrea gigas (Thunberg)] were examined using latroscan thin-layer chromatography/flame-ionization detection (TLC/FID), and fatty acid compositions determined by capillary gas chromatography and gas chromatography/mass spectrometry (GC/MS). The fatty acid methyl esters were separated using argentation TLC and also analyzed as their 4,4-dimethyloxazoline derivatives using GC/MS. Major esterified fatty acids inC. gigas were 16∶0, 20∶5n−3, and 22∶6n−3. C₂₀ and C₂₂ nonmethylene interrupted (NMI) fatty acids comprised 4.5 to 5.9% of the total fatty acids. The NMI trienoic fatty acid 22∶3(7,13,16) was also identified. Very little difference was found in the proportions of the various lipid classes, fatty acids or sterols between samples of adult oysters of two different sizes. However, significant differences in some of the lipid components were evident according to the method of sample preparation used prior to lipid extraction with solvents. Lyophilization (freeze drying) of samples led to a significant reduction in the amounts of triacylglycerols (TG) extracted by solvents in two separate experiments (7.0 and 52.5% extracted). Extracts from lyophilized samples had less 16∶0, C₁₈ unsaturated fatty acids, and 24-ethylcholest-5-en-3β-ol, while C₂₀ and C₂₂ unsaturated fatty acids comprised a higher proportion of the total fatty acids. There was no significant change in the amounts of polar lipids, total sterols, free fatty acids or hydrocarbons observed in extracts from lyophilized samples relative to extracts from nonlyophilized samples. Addition of water to the freezedried samples prior to lipid extraction greatly improved lipid yields and resulted in most of the TG being extracted.     

Molecular analysis of intact preen waxes of Calidris canutus (Aves: Scolopacidae) by gas chromatography/mass spectrometry

The intact preen wax esters of the red knot Calidris canutus were studied with gas chromatography/mass spectrometry (GC/MS) and GC/MS/MS. In this latter technique, transitions from the molecular ion to fragment ions representing the fatty acid moiety of the wax esters were measured, providing additional resolution to the analysis of wax esters. The C₂₁−C₃₂ wax esters are composed of complex mixtures of hundreds of individual isomers. The odd carbon-numbered wax esters are predominantly composed of even carbon-numbered n-alcohols (C₁₄, C₁₆, and C₁₈) esterified predominantly with odd carbon-numbered 2-methyl fatty acids (C₇, C₉, C₁₁, and C₁₃), resulting in relatively simple distributions. The even carbon-numbered wax esters show a far more complex distribution due to a number of factors: (i) Their n-alcohol moieties are not dominated by even carbon-numbered n-alcohol moieties are not dominated by even carbon-numbered n-alcohols esterified with odd carbon-numbered 2-methyl fatty acids, but odd and even carbon-numbered n-alcohols participate in approximately equal amounts; (ii) odd carbon-numbered methyl-branched alcohols participate abundantly in these wax ester clusters; and (iii) with increasing molecular weight, various isomers of the 2,6-, 2,8-, and 2,10-dimethyl branched fatty acids also participate in the even carbon-numbered wax esters. The data demonstrate that there is a clear biosynthetic control on the wax ester composition although the reasons for the complex chemistry of the waxes are not yet understood.     

Selective Microbial Degradation of Saturated Methyl Branched-Chain Fatty Acid Isomers

Three strains of Pseudomonas (P.) bacteria were screened for their capabilities of degrading chemically synthesized saturated branched-chain fatty acids (sbc–FA). Mixtures of sbc–FA with the methyl-branch located at various locales along the fatty acid were used as a carbon feedstock in shake-flask culture. Utilization (and hence degradability) of the sbc–FA was monitored based on positive bacterial growth, fatty acid recovery rates and chromatographic (gas chromatography (GC) and GC-mass spectroscopy (MS)) analysis of the recovered carbon source. P. putida KT2442 and P. oleovorans NRRL B-14683 were both able to grow on sbc–FA utilizing 35 wt% and 27 wt% of the carbon source, respectively after 144 h. In contrast, P. resinovorans NRRL B-2649 exhibited the most efficient use of the carbon source by utilizing 89 % of the starting material after 96 h resulting in a cell dry weight (CDW) of 3.1 g/L. GC and GC–MS analysis of the recovered carbon source revealed that the bacterial strains selectively utilized the isostearic acid in the sbc–FA mixture, and a new group of C₁₀, C₁₂, C₁₄ and C₁₆-linear and/or branched-chain fatty acids (approximately 4–29 wt%) were formed during degradation.     

Chromatograpahic Analysis of Eriobotrya japonica Fat

Thin layer chromatography and infrared spectophotometry of the fat of Eriobotrya japonica shows the presence of hydrocarbons (3.1%), was esters (5.3%), triglycerides (786%) and the last polar fraction containing fatty acids, colouring matter etc. (13.0%). After the saponification of the fat, the fatty acids C₁₂-C₂₄ and fatty alcohols C₁₂-C₂₆ determined first time as methyl esters and acetates respectively were identified by the application of gas liquid chromatography.     

Characterization of skin-surface lipids from the monkey (Macaca fascicularis)

Skin-surface lipids from the monkeyMacaca fascicularis are composed of sterol esters (38%), cholesterol (4%) and two types of wax diesters, identified as Type II (IIa and IIb, 17% and 40%, respectively). Type IIa contained diesters of 1,2-alkanediols esterified with two molecules of long-chain (C₁₄−C₃₄) fatty acids having straight and branched chains. In the diesters IIa, fatty acids shorter than C₁₉ predominated in position 1, and fatty acids longer than C₂₀ predominated in position 2. Type IIb contained diesters of 1,2-alkanediols esterified with C₄ and C₅ branched-chain fatty acids (predominantly isovaleric acid) at position 1 and long-chain (C₁₄−C₂₇) acids, having straight and branched chains, at position 2. The shortchain acids were converted to 2-nitrophenylhydrazides and analyzed by high-performance liquid chromatography (HPLC). Ammonia chemical ionization (CI)-gas chromatography (GC)-mass spectrometry (MS) resolved the intact diesters IIb into 12 peaks corresponding to molecular weights ranging from 597 to 748, and showed that the molecular species, such as C₂₁−C₁₆−C₅ (diol, fatty acid in position 2, fatty acid in position 1), C₂₂−C₁₆−C₅ and C₂₃−C₁₆−C₅, were prevalent. The fatty acids from both diesters were mostly (>98%) saturated. The 1,2-alkanediols from both diesters consisted of C₁₆−C₂₆ saturated straight- and branched-chain components. The acyl groups of sterol esters contained 86% C₁₄−C₃₄ branched-chain acids. The unsaturated fatty acids (5.4%) belonged to a straight-chain monoenoic series having extremely long chains (C₁₈−C₃₄). The branched-chain structures in the fatty acids and diols were iso and anteiso. These results show the species-specific profile for the skin-surface lipid synthesis.     

The steryl ester and phospholipid fatty acids of the spongeAgelas conifera from the Colombian Caribbean

The steryl ester and phospholipid fractions of the marine spongeAgelas conifera were isolated and analyzed. The fatty acyl components of the steryl ester and phospholipid fractions as determined by gas chromatography and gas chromatography/mass spectrometry were very similar and consisted of 56.8 and 62.7% of C₁₄−C₂₀ acids (normal; branched, especiallyiso andanteiso; and monounsaturated, particularly Δ⁹ and Δ¹¹ acids) and of 43.1 and 35.5% of C₂₄−C₂₆ acids (Δ^5,9 diunsaturated acids), respectively. The major constituent fatty acids detected were 13-methyltetradecanoic,n-hexadecanoic, 10-methylhexadecanoic, 11-octadecenoic, 12-methyloctadecanoic, 5,9-pentacosadienoic and 5,9-hexacosadienoic acids. The phospholipids isolated were identified as phosphatidylcholine (37%), phosphatidylserine (34%), phosphatidylethanolamine (16%) and phosphatidylinositol (11%). The distribution of fatty acids within the phospholipid classes was also determined.     

Up to 21 Different Sulfur-Heterocyclic Fatty Acids in Rapeseed and Mustard Oil

Three sulfur-heterocyclic fatty acids (SHFA) had been tentatively identified in rapeseed oil in the late 1980s. In this study we aimed to enrich and verify the presence of potential SHFA in one sample of native rapeseed oil, refined rapeseed oil and mustard seed oil. Fifty-gram samples of the three oils were individually saponified and converted into methyl esters. The resulting samples were hydrogenated and subjected three times to urea complexation. The resulting extracts of native rapeseed oil and mustard oil contained 21 different SHFA with 18, 20, 22 or 24 carbons. The refined rapeseed oil contained only nine C₁₈-SHFA. Structure investigation of the SHFA was performed by gas chromatography with mass spectrometry (GC/MS) using methyl esters and also 3-pyridylcarbinol esters. A direct screening of non-enriched samples by GC/MS in the selected ion monitoring mode and by GC with flame photometric detector (sulfur-selective) verified that the SHFA were native compounds of the oils and no artefacts of the sample preparation. Similar abundances of the four isomer groups of SHFA with monoenoic fatty acids of the same carbon number in these and five further rapeseed and mustard samples indicated that these could be the precursors of the SHFA.     

Characterization and Authentication of Significant Chinese Edible Oilseed Oils by Stable Carbon Isotope Analysis

The ratios of stable carbon isotopes (δ¹³C) of 12 oils extracted from Chinese edible oilseed samples and their individual fatty acids were determined by elemental analysis-isotope ratio mass spectrometry (EA-IRMS) and gas chromatography-isotope ratio mass spectrometry (GC-IRMS). The results have demonstrated that the δ¹³C ratios of the oils from C3-plant seeds range from ?26.8 to ?30.7‰, while the δ¹³C ratios of C4-plant maize oil are in the interval of ?14.1 to ?16.2‰. Eighteen fatty acids were identified and their abundances were measured by gas chromatography–mass spectrometry (GC–MS) in these oils with C_16:0, C_18:0, C_18:1 and C_18:2 as the major constituents. From the data on fatty acids and stable carbon isotopes, several sensitive markers were developed to detect the adulteration of Chinese edible oilseed oils. Examples are provided with pre-blended samples to illustrate the discrimination procedures and corresponding sensitive markers with emphasis on camellia seed oil, flax seed oil and perilla seed oil.     

Linear, steroidal, and triterpene esters, and steryl glycosides from Festuca argentina

Ester waxes and steryl glycosides of the grass Festuca argentina were studied. Saponification of the waxes from the petroleum ether extract led to n-hexacosanol as the major single linear alcohol, along with pentacyclic triterpenols, such as β-amyrin, germanicol, isobaurenol, lupeol, hopenol-a and hopeol, and low amounts of sterols, such as cholesterol, campesterol, stigmasterol, sitosterol and dihydrositosterol, identified by gas chromatography/mass spectrometry (GC/MS). Fatty acids were identified as methyl esters as C_12∶0, C_14∶0, C_16∶0, C_18∶0, C_18∶2, and C_20∶0. The occurrence of a wide chainlength range of fatty acids and a single linear alcohol closely matched for other reports on the tribe Festuceae. On the contrary, pentacyclic triterpenols with a variety of skeletons, especially isobauerenol, are not usual as esters of fatty acids in the Gramineae. Low amounts of steryl glycosides were also obtained from the methylene chloride percolate of the methanol extract. Upon acetylation followed by hydrolysis, aglycones were identified by capillary gas-liquid chromatography (GLC) and GC/MS. As Δ⁷-cholesterol, campesterol, stigmasterol, sitosterol, dihydrositosterol, and the sugars as glucose, xylose, and arabinose by GLC of the respective alditol acetates. This is the first report on the linear, steryl, and triterpenyl esters of F. argentina. It is noteworthy that Δ⁷-steryl glycosides are rare, and steryl monoarabinosides have not been proviously reported on the family Gramineae.     

Identification of Unique Aldehyde Dimers in Sorghum Wax Recovered after Fermentation in a Commercial Fuel Ethanol Plant

Sorghum wax can be extracted from the surface of sorghum (Sorghum bicolor) kernels. It is composed mostly of a mixture of unsaturated C₂₈ and C₃₀ alkanes, fatty acids, fatty alcohols, and fatty aldehydes. Like carnauba wax, sorghum wax is a hard wax with a high melting point and it has potential edible and industrial applications. The yield of sorghum wax from the surface of sorghum kernels is 0.2–0.5 g of wax per 100 g of kernels. Sorghum wax can also be recovered from the “distillers oil” which is obtained after fermentation of sorghum (milo) or sorghum/corn blends in dry grind fuel ethanol plants. This distillers sorghum wax can potentially be obtained in yields of up to 10% by chilling the distillers oil to precipitate the wax and then recovering it via centrifugation or filtration. Like sorghum kernel wax, distillers sorghum wax is mainly composed of C₂₈ and C₃₀ alkanes, alcohols, and aldehydes in the molecular weight (MW) range of 350–450. However, we found that 7–49% w/w of distillers sorghum wax is composed of larger wax components with MW of 799–912. Analysis via high-resolution atmospheric pressure chemical ionization mass spectrometry (APCI) and gas chromatography with electron ionization mass spectrometry (GC/MS-EI) resulted in exact mass data and fragmentation patterns that suggested that these high MW compounds are monounsaturated fatty aldehyde dimers, likely formed by aldol condensation. Further confirmation supporting the GC/MS data for the aldol reaction was obtained by comparison with similar aldol products.     

Sponge fatty acids. 3. Occurrence of series of n−7 monoenoic andiso-5,9 dienoic long-chain fatty acids in the phospholipids of the marine spongeCinachyrella aff.schulzei keller

The fatty acid composition of phospholipids from the New Caledonian spongeCinachyrella aff.schulzei Keller was studied. More than 60 fatty acids were identified as methyl esters andN-acyl pyrrolidides by gas chromatography and gas chromatography/mass spectrometry. Two isoprenoid fatty acids also were shown to be present, namely 4,8,12-trimethyltridecanoic and 5,9,13-trimethyltetradecanoic acids. The unusual 6-tetradecenoic, 6-pentadecenoic, 12-nonadecenoic and 26-methylheptacosanoic (iso-28∶0) acids were found for the first time in sponge phospholipids. A series of six n−7 monoenoic long-chain fatty acids (C₂₃ to C₂₈) were identified, including the rare 16-tricosenoic, 18-pentacosenoic and 21-octacosenoic acids. Fifteen fatty acids possessing the typical 5,9 dienoic moiety accounted for 30% of the total fatty acid mixture. Two new fatty acids were identified, namely 5(Z)-octacosenoic and 27-methyl-5(Z),9(Z)-octacosadienoic (iso-5,9-29∶2). Based on gas chromatography/Fourier transform infrared experiments, the double bonds were assigned the (Z) configuration. For part 2 of this series, see Reference 1.     

Quantitation of acyl migration during lipase-catalyzed acidolysis, and of the regioisomers of structured triacylglycerols formed

Various MLM-type (M, medium-chain fatty acids; L, long-chain fatty acids) structured triacylglycerols were produced in pilot- or small-scale packed-bed reactors by lipasecatalyzed acidolysis. The incorporation and acyl migration of octanoic acid were measured by gas chromatography and Grignard degradation, and ranged from 39.0 to 48.7% and 0.6 to 9.3%, respectively. Quantitation of triacylglycerol molecular species was performed by ammonia negative ion chemical ionization (NICI) mass spectrometry (MS). The proportion of ACN (acyl carbon number) 34 species that contained one C₁₈ fatty acid and two C_8∶0′ in samples analyzed, varied from 12.5 to 23.2%. The selected regioisomers MLM and MML within the ACN 34 species group were quantified by NICI tandem MS (MS/MS) and were in the range of 97.1 to 98.4% and 1.6 to 2.9%, respectively. There was no correlation between the level of acyl migration during lipase-catalyzed esterification and the level of regioisomers of the selected MLM-type triacylglycerols in the structured lipid samples.     

Isolation,characterization, and antifungal application of a biosurfactant produced by Enterobacter sp. MS16

Isolate MS16 obtained from diesel contaminated soil, identified as Enterobacter sp. using 16S rRNA gene analysis produced biosurfactant when grown on unconventional substrates like groundnut oil cake, sunflower oil, and molasses. Of these carbon substrates used, sunflower oil cake showed highest biosurfactant production (1.5 g/L) and reduction in surface tension (68%). The biosurfactant produced by MS16 efficiently emulsified various hydrocarbons. The carbohydrates and fatty acids of the biosurfactants were studied using TLC, FTIR, NMR, and GC‐MS. The carbohydrate composition as determined by GC‐MS of their alditol acetate derivatives showed the predominance of glucose, galactose and arabinose, and hydroxyl fatty acids of chain length of C₁₆ and C₁₈ on the basis of FAMEs analysis. Biosurfactant showed antifungal activity and inhibited the fungal spore germination. Practical applications : Enterobacter sp., MS16 produces a biosurfactant composed of carbohydrates and fatty acids which exhibits excellent surface active properties. Use of industrial wastes for biosurfactant production is economical and facilitates the industrial production of this biosurfactant which has potential antifungal activity.     

Gas chromatographic resolution of butteroil and synthetic triglycerides beyond their carbon numbers

Preparative gas chromatography was used to isolate the C₂₆ to C₃₈ triglycerides of butteroil in groups of uniform molecular weight. Determination of fatty acids allowed a preliminary assignment of triglyceride structure and an estimation of the relative proportions of the various glyceride types. A basis for a further gas chromatographic resolution of saturated triglycerides of identical molecular weight but differing in composition and positional distribution of fatty acids was recognized in the easy separation of various derivatives of 1- and 2-monoglycerides. The study indicated that it might be practical to separate positional isomers of those triglycerides which contain fatty acids of both short and long chain lengths. Other promising routes to increased glyceride resolution were suggested by experiments using chromatographic conditions approaching gas-solid systems.     

Compositional and thermal characterization of genuine and randomized lard: A comparative study

Composition and thermal characterization of genuine and randomized lard were investigated comparatively in an attempt to find common merits that assess lard detection. The investigation included compositional and positional distribution of fatty acids, triacylglycerol profiling by gas chromatography (GC) and reversed-phase high-performance liquid chromatography (RP-HPLC), as well as thermal behavior by differential scanning calorimetry (DSC) of both samples. Individual and total saturated and unsaturated fatty acid composition in total fats of both genuine and randomized lard were identical. On the other hand, the results of pancreatic lipolysis/GC analysis showed that the average percent palmitic acid [PAEF(%)] and myristic acid [MAEF(%)] enrichment factors of genuine (280 and 270) and randomized lard (110 and 98) were quite different. Thus, application of PAEF to detect randomized lard is of no value. However, normalization of fatty acid distribution by randomization in 2-monoacylglycerols made the individual and total saturated and unsaturated fatty acids almost identical to that of total fat and neutral triacylglycerols (TG) of lard. TG compositional analysis by GC revealed that both genuine and randomized lard had six dominant TG (C_46′C_48′C_50′C_52′C_54′ and C₅₆) with quite different concentrations. TG with C₅₂ represent the major constituent of genuine and randomized lard. TG profiling of samples was also carried out by RP-HPLC with a refractive index detector. The same peaks were eluted in both samples, but the area % of major peaks changed due to randomization. 2-Palmitooleostearin (SPO) was found in high proportion in lard. However, the ratios of SPO to 2-palmitooleolinolein of both genuine and randomized lard are close (0.6±0.05) and significantly distinguishable from that of beef (4.24), mutton (6.17), chicken (0.21), and turkey (0.14) fats. The DSC thermogram and thermodynamics of phase transitions of both samples were quite different and do not reveal common characteristic(s) that could be used for immediate detection of lard substances in fat admixtures.     

Molecular species of glycerophospholipids and sphingomyelins of human erythrocytes: Improved method of analysis

This study reports the application of modern methods of molecular species analysis in determination of the structure of both major and minor glycerophospholipids and sphingomyelins of human erythrocytes. Individual phospholipid classes were resolved from total lipid extracts by thin-layer chromatography. Diradylglycerols were released by phospholipase C and converted into trimethylsilyl ethers, which were resolved into the alkenylacyl, alkylacyl and diacylglycerol subclasses by normal phase high performance liquid chromatography. Molecular species of diradylglycerols and ceramides were quantitated according to carbon and double bond number by gas liquid chromatography using a fused silica capillary column wall-coated with bonded RTx-2330. The molecular species of ceramides were determined by GC/MS. The diradyl glycerophosphocholines contained 93.0% diacyl, 4.6% alkylacyl and 2.5% alkenylacyl, white the diradyl glycerophosphoethanolamines were made up of 48.8% diacyl, 47.8% alkenylacyl and 3.4% alkylacyl subclasses. Analysis of the molecular species showed that the long chain polyunsaturated acids were mainly combined with C₁₆ in all diradyl GPC subclasses and in diacyl GPE, while in the alkylacyl and alkenylacyl GPE and in diacyl glycerophosphoinositol and diacyl glycerophosphoserine they were combined mainly with C₁₈ saturated fatty chains. In addition to the C₁₆ and C₁₈ alkyl and alkenyl, the ether fractions also contained significant proportions of C₂₀, C₂₂ and C₂₄ chains. The molecular species of the ceramide moieties of the SPH were made up largely of mono- and diunsaturated species. Over 200 molecular species were identified and quantitated in a representative sample of human red blood cells.     

Stable isotope characterization of olive oils. I—Compositional and carbon-13 profiles of fatty acids

Nearly 200 olive oils produced in the Mediterranean basin, mainly in Greece, during 4 yr from 1993 to 1996, were studied by gas chromatography (GC) and on-line GC-isotope ratio mass spectrometry (GC-C-IRMS). The composition of the oils in the more abundant fatty acids (C_16:0, C_16:1, C_18:0, C_18:1, C_18:2, and C_18:3) was obtained by GC after transesterification of the triglycerides into methyl esters. Using the hyphenated GC-C-IRMS technique, the ¹³C contents of the three most abundant acids, C_16:0, C_18:1, and C_18:2, were measured with satisfactory accuracy. The results, analyzed in terms of geographical, temporal, and botanical factors, provide new criteria for the authentication of olive oils.     

Postparturition changes in the triacylglycerols of cow colostrum

The changes in the triacylglycerol (TAG) composition of colostrum fat of three cows were studied. In addition to the determination of fatty acid composition by gas chromatography, the distribution of TAG according to the acyl carbon number (ACN) and molecular weight was analyzed utilizing both supercritical fluid chromatography (SFC) and ammonia negative-ion chemical ionization mass spectrometry (MS). Colostrum TAG contained substantially less stearic and oleic acids and more myristic and palmitic acids than the normal Finnish milk fat. The major trends in the changes of fatty acids and TAG were similar for each cow, although clear differences between individuals were found. During the first week of parturition, the proportions of short-chain fatty acids (C₄–C₁₀) typically increased as well as those of stearic and oleic acids, whereas the relative amounts of C₁₂–C₁₆ acids decreased, especially those of myristic and palmitic acids. Distinct changes occurred also in TAG distributions: the proportions of molecules with ACN 38–40 increased and those with ACN 44–48 decreased. Although there were distinct differences between individuals shortly after delivery, both the fatty acid compositions and TAG distributions of the milk samples of the cows started to resemble each other after one week. The theoretical profiles of colostrum TAG calculated based on the fatty acid compositions differed clearly from the ACN distributions analyzed by SFC and MS. Thus, the analysis of TAG is essential, because the changes in molecular species composition of colostrum TAG cannot be estimated according to the fatty acid analysis alone.     

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.     

Occurrence ofcis-5,cis-9-Hexacosadienoic andcis-5,cis-9,cis-19-Hexacosatrienoic acids in the marine spongeMicrociona prolifera

Fatty acid analysis of the total lipids from the marine spongeMicrociona prolifera by gas liquid chromatography on an EGSS-X column revealed two major peaks with equivalent chain length values of 27.08 and 27.74. Each of these components was isolated as a separate band by thin layer chromatography on AgNO₃-silicic acid. Characterization of the two unknowns by IR spectroscopy, NMR, hydrogenation, and gas liquid chromatography revealed that the unknown acids weren-26∶2 andn-26∶3 containing only nonmethylene interruptedcis-double bonds. Reductive ozonolysis identified the 26∶2 ascis-5,cis-9-hexacosadienoic acid and the 26∶3 ascis-5,cis-9,cis-19-hexacosatrienoic acid. Analysis of the fatty acid composition ofMicrociona total lipids showed 14% 26∶2 and 31% 36∶3. The neutral lipids, phosphatidylethanomaline, and phosphatidylserine all contained >41% C₂₆ acids; but only 4% C₂₆ was present in the phosphatidylcholine.