Über die Zusammensetzung des Samenöles aus Traubenholunder (Sambucus racemosa L.)

Composition of the Seed Oil of Clustered Elder (Sambucus racemosa L.) The seeds of clustered elder (Sambucus racemosa L.) contain 28% of an oil. Its separation to groups of components was carried out by adsorption chromatography. Single groups of substances were further analysed by TLC and GLC in combination with physico-chemical methods (IR and MS). Thus the following substances were found and identified: n-alkanes (C₁₇–C₃₃), 2-methyl- and 3-methylalkanes, squalene, a mixture of esters, triglycerides, β-sitosterol, campesterol and a mixture of diglycerides. The conditions for the separation of esters by silica gel column chromatography were also found. The mixture of esters was thus separated into a group of aliphatic wax esters (C₃₄–C₄₄) and four groups of esters of steroidal alcohols (mainly β-sitosterol and campesterol) with aliphatic unsaturated acids (predominantly C_18:1, C_18:2 and C_18:3). In the triglycerides representing the main fraction of the oil (93%), the acids C_16:0, C_18:1, C_18:2 and C_18:3 are present. The same acids were also found in diglycerides.     

Molecular species of wax esters in jaw fat of atlantic bottlenose dolphin,Tursiops truncatus

The jaw fat of the Atlantic bottlenose dolphin (Tursiops truncatus) contains unusual wax esters which can be separated into short chain (<C₂₄) and long chain (>C₂₄) fractions by thin layer chromatography. The short chain wax esters (28 wt. %) have been characterized as a 72∶24∶4 mixture of isovaleroyl, isobutoryl, and 2-methylbutyrol, esters of C₁₄–C₁₈ n- and iso-alcohols. The intact <C₂₄ esters have been resolved into individual molecular species by gas liquid chromatography on open-tubular polyester columns. The long chain wax esters (12 wt. %) contain C₁₀–C₂₂ n- and iso-acids esterified to the same C₁₄–C₁₈ n- and iso-alcohols. Gas liquid chromatography of the intact, hydrogenated >C₂₄ esters on a short JXR column has characterized them according to carbon number and the number of methyl branches they contain.     

Biochemical and physiological studies of certain ticks (Ixodoidea): Isolation and partial identification of a new fatty acid in eggs ofDermacentor andersoni Stiles (Ixodidae)

Gas liquid chromatographic analysis of the fatty acid methyl esters from eggs ofDermacentor andersoni Stiles (Ixodidae) revealed the presence of significant quantities (15% total fatty acids) of an unidentified component with a retention time between C_18∶3−C_22∶0 fatty acids. Smaller amounts of the unidentified component (ca. 5% total fatty acid) also were detected in host rabbit serum. Purified, the unidentified component's methyl ester collected from the tick eggs by preparative gas liquid chromatography was partially identified and characterized by chemical and spectroscopic analyses. The evidence suggests that the unidentified component is a methyl branched C₁₅ tricarboxylic acid containing two vicinal carboxylic acid groups. Biosynthesis of the unidentified component by the tick is under investigation.     

Studies of waxes. VI. The n-acids of carnauba wax

Summary The proportion of normal acids among the acids of carnauba wax has been found, by a chromatographic separation method, to be 38%. The normal acid mixture has been separated into its components by amplified distillation of the methyl esters, which revealed the presence of the acids of even carbon number from C₁₈ to C₃₀. The acids were identified by their melting points, long crystal spacings, and crystal habit. An estimate of their relative amounts is as follows: C₁₈, 3%; C₂₀, 11.5%; C₂₂, 9%; C₂₄, 30%; C₂₆, 12%; C₂₈, 16.5%; C₃₀, 7%. Part V of this series: The Aliphatic Alcohols of Wool Wax, by K. E. Murray and R. Schoenfeld, J. Am. Oil Chem. Soc.,29; 416–420 (1952).     

Wax ester fraction of edible oils: Analysis by on‐line LC‐GC‐MS and GC×GC‐FID

The wax ester fraction of various plant oils was isolated by normal‐phase HPLC (NPLC) on‐line coupled to GC via the on‐column interface and applying concurrent eluent evaporation. The esters were analyzed by on‐line NPLC‐GC‐MS and by comprehensive two‐dimensional GC with flame ionization detection (GC×GC‐FID) off‐line combined with NPLC‐GC. GC×GC‐FID enables to group the various classes of wax esters, in particular the phytol esters, geranylgeraniol esters and the straight‐chain esters of palmitic acids and the unsaturated C₁₈ acids. Optimization of the GC×GC columns and the conditions must take into account the limited thermostability of the diterpene esters. Chromatograms are shown for a range of oils, with particular focus on the various classes of wax esters in olive oil and the geranylgeraniol esters 22:0 and 24:0 in a variety of oils.     

Preparation of alcohols from cyclic fatty acids

Saturated C_18- and C₂₀-cyclic alcohols have been prepared by catalytic hydrogenation of methyl esters from cyclized linseed monomeric acids, purified saturated C₁₈-cyclic acids, ethylene adduct of conjugated soybean fatty acids, and ethylene adduct of conjugated octadecadienoic acids. The cyclic alcohols have also been prepared from free acids of crude cyclic linseed, cyclic linseed monomeric, and ethylene adduct of 9,11,t,t,-octadecadienoic. Conversion of esters and acids was 88–99% by hydroxyl determination; by gas-liquid chromatographic analysis, almost quantitative Hydrogenations were carried out with 10%, by weight, copper chromite catalyst, an initial hydrogen pressure of 2,100 psi, and a temperature of 280C for 3–5 hr. Preliminary evaluations indicate that saturated C_18- and C₂₀-cyclic alcohols have a potential use in cosmetic formulations. Presented AOCS meeting in Minneapolis, Minn., 1963. No. Utiliz. Res. & Dev. Div., ARS, USDA.     

Fatty acids,fatty alcohols,wax esters,and methyl esters fromCrambe abyssinica andLunaria annua seed oils

Crambe abyssinica andLunaria annua, members of the Cruciferae family, have seed oil glycerides containing ca. 55–65% of C₂₂ and C₂₄ unsaturated fatty acids. Fatty acids were prepared by saponification; fatty alcohols, by sodium reduction of glycerides; liquid wax esters, byp-toluenesulfonic acid-catalyzed reaction of fatty acids with fatty alcohols; and methyl esters, by reaction of fatty acids with diazomethane. Solid hydrogenated glyceride oils and wax esters were compared with several commercial waxes. Chemical and physical constants were determined for the seed oils and their derivatives. Position of unsaturation in theCrambe fatty acids was determined by gas chromatographic analysis of the permanganate-periodate degradation products. The major dicarboxylic acid was brassylic (C₁₃), proving the docosenoic acid to be erucic. Presented in part at the AOCS meeting in New Orleans, La., 1962. A laboratory of the No. Utiliz. Res. & Dev. Div., ARS, U.S.D.A.     

Structure-retention correlation of isomeric bile acids in inclusion high-performance liquid chromatography with methyl β-cyclodextrin

The structure-retention correlation of various C₂₄ bile acid isomers was studied by the addition of methyl β-cyclodextrin (Me-β-CD) to mobile phases in reversed-phase high-performance liquid chromatography (HPLC). The compounds examined include a series of monosubstituted bile acids related to cholanoic acids differing from one another in the position and configuration of an oxygen-containing function (hydroxyl or oxo group) at the position C-3, C-6, C-7, or C-12 and the stereochemistry of the A/B-ring fusion (trans 5α-H and cis 5β-H) in the steroid nucleus. The inclusion HPLC with Me-β-CD was also applied to biologically important 4β- and 6-hydroxylated bile acids substituted by three to four hydroxyl groups in the 5β-steroid nucleus. These bile acid samples were converted into their fluorescence prelabeled 24-pyrenacyl ester derivatives and chromatographed on a Capcell Pak C₁₈ column eluted with methanol-water mixtures in the presence or absence of 5 mM Me-β-CD. The effects of Me-β-CD on the retentions of each compound were correlated quantitatively to the decreasing rate of capacity factors and the relative strength of host-guest inter-actions. On the basis of the retention data, specific and nonspecific hydrogen-bonding interactions between the bile acids and the Me-β-CD were discussed.     

Fatty Acid Specificity of Candida cylindracea Lipase

Candida cylindracea lipase (SIGMA) was tested against triglycerides (TG) and wax esters (WE) of marine origin as substrates. Under the same conditions, wax esters were hydrolysed at a lower rate than the triglycerides. The C₁₄ to C₁₈ saturated and monounsaturated fatty acids were preferentially hydrolysed whereas the longer chain monoenes (20:1 and 22:1) and particularly the polyunsaturated fatty acids (18:4,20:5 and 22:6) were resistant to the hydrolysis in triglycerides as well as in wax esters. No specificity was demonstrated for the fatty alcohols in the wax esters.     

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.     

Long-chain fatty acids containing ether linkage. I. The antibacterial and fungicidal activities of some new β-alkyloxypropionic acids and their methyl esters

β-Alkoxypropionic acids and their methyl esters were made with alkoxy groups ranging from C₄H₉O to C₁₈H₃₅O: R-O-CH₂CH₂COOH (CH₃). Methyl esters and acids were also made with one and with two oxyethylene groups between the alkoxy group and the propionic acid group: RO (CH₂ CH₂ O) n-CH₂CH₂ COOH(CH₃). The compounds were tested againstStaphylococcus aureus and againstPenicillium for growth inhibition. The optimum size of the alkoxy group appears to be R=C₁₂H₂₅. Oxyethylene groups enhanced the activity againstS. aureus, but had relatively little effect againstPenicillium.     

Occurrence of wax esters and 1-O-alkyl-2,3-diacylglycerols in goat epididymal sperm plasma membrane

Two unusual lipid classes were detected by thin-layer chromatography in the neutral lipids derived from goat cauda-epididymal sperm plasma membrane. The lipids were identified as wax esters and 1-O-alkyl-2,3-diacylglycerols based on chromatographic properties, identity of their hydrolysis products, and infrared/¹H nuclear magnetic resonance spectral evidence. The membrane containedca. 3 and 5 μg/mg protein of wax esters and alkyldiacylglycerols, respectively. The relative proportions of wax esters and alkyldiacylglycerols in the total neutral lipids were 1.5% and 2.4%, respectively. The lipids contained fatty acids with chain lengths of C₁₄ to C₂₂. The major fatty acids of the wax esters were 14∶0, 16∶0, 16∶1ω7, 18∶0 and 18∶1ω9. The fatty acids in alkyldiacylglycerol were 16∶0, 18∶0, 22∶5ω3 and 22∶6ω3. Alkyldiacylglycerol was particularly rich in docosahexaenoic acid 22∶6ω3) representing 30% of the total fatty acids. The alcohols of wax ester were all saturated with C₂₀–C₂₉ carbon chains. The deacylated products derived from alkyldiacylglycerols were identified as hexadecyl, octadecyl and octadec-9′-enyl glycerol ethers.     

Component fatty acids of marine fish liver oils

Summary 1. Two liver oils (Elasmobranch) fromCarcharias melanopterus andPristis cuspidatus, caught off the Madras coast are studied, and their component fatty acids are reported. 2. The mixed acids were separated into three groups (varying unsaturation) of acids, and their methylesters were fractionated. 3. The liver oils are found to belong to the fourth group of Tsujimoto’s classification of Elasmobranch fish liver oils.Carcharias melanopterus liver oil contains 31.1% unsaturated acids (myristic 3.1, palmitic 18.4, stearic 9.5, and 0.1% arachidic) and 68.9% unsaturated acids (C₁₆ 10.8, C₁₈ 19.7, C₂₀ 15.2, C₂₂ 17.1, C₂₄ 5.3%, and traces of C₁₄ monoethenoid).Pristis cuspidatus liver oil contains 36.9% saturated acids (myristic 1.2, palmitic 22.9, stearic 12.7, and arachidic 0.1%) and 67.1% unsaturated acids (C₁₆ 8.2, C₁₈ 28.5, C₂₀ 16.4, C₂₂ 5.2, C₂₄ 4.6%, and traces of C₁₄ monoethenoid). The unsaturations of the different groups of acids are almost of the same order. 4. The abnormal content of saturated acids can be explained by the process of bio-hydrogenation. The relatively less amount of saturated acids inCarcharias melanopterus liver oil along with its higher content of polyethylenic acids (C₂₀ and above) points strongly to the possible presence of intermediate types of fats among the four groups of Elasmobranch oils.     

Cuticular lipids of insects: V. Cuticular wax esters of secondary alcohols from the grasshoppersMelanoplus packardii andMelanoplus sanguinipes

Wax esters of secondary alcohols constitute 18–20% of the cuticular lipid extract ofMelanoplus packardii and 26–31% of the cuticular lipids ofMelanoplus sanguinipes. The total number of carbons in the wax esters range from 37–54 with 41 predominating in both species. The fatty acids ofM. packardii wax esters are 16∶0, 18∶0, 14∶0, 20∶0 and 12∶0 in decreasing quantity. The fatty acids ofM. sanguinipes wax esters are 18∶0, 20∶0, 16∶0 22∶0, 14∶0, 19∶0 and 17∶0 in decreasing quantity. The secondary alcohols from the wax esters ofM. packardii are C₂₅, C₂₃ and C₂₇ in decreasing quantity, and the secondary alcohols of theM. sanguinipes are C₂₃, C₂₅, C₂₁, C₂₇, C₂₄, C₂₂ and C₂₆ in decreasing quantity. Each secondary alcohol consists of two to four isomers with the hydroxyl group located near the center of the chain. Montana Agriculture Experiment Station, Journal Series No. 332.     

Enzymatic synthesis of medium‐chain triacylglycerols by alcoholysis and interesterification of copra oil using a crude papain lipase preparation

We have examined the possibility of producing analogs of medium‐chain triglycerides (MCT) from copra oil, i.e. a triacylglycerol mixture with a high content of medium‐chain fatty acid moieties (C₆–C₁₀). A two‐step enzymatic process was used in which copra triacylglycerols were first split with papain lipase by alcoholysis with an alkyl alcohol and then subjected to interesterification with the alkyl esters recovered using papain lipase. Effects of temperature, water activity content, substrate ratio, biocatalyst amount, and alcohol chain length were also investigated. On the one hand, the sn‐3 stereoselectivity of the lipase in the alcoholysis of copra oil with butanol has permitted a direct enrichment of caproic, caprylic and capric moieties in the synthesized butyl esters. Thus, in the batch reactor, the reaction led to about 31% conversion of the oil after 24 h, and the content of C₆–C₁₀ acids in the synthesized esters increased from about 16% in the starting oil to almost 42%. A similar enzymatic alcoholysis in a packed‐bed column bioreactor gave 31% conversion of the oil after 120 min of reactor residence time. The reaction was also very selective because the C₆–C₁₀ fatty acyl groups represented about half of the newly formed butyl esters, whereas they accounted for only 16% of total fatty acids in the starting oil. On the other hand, the transesterification of the alkyl esters recovered (highly enriched in C₆–C₁₀ fatty acyl groups) with native copra oil directly led to an increase in the content of MCT in the oil, from 18 mol‐% at the beginning of the reaction to 61 mol‐% of MCT after a time period of 72 h in the batch reactor.     

Quantitative analysis of long-chain trans-monoenes originating from hydrogenated marine oil

Gas chromatography (GC) is used for the analysis of trans-fatty acids in partially hydrogenated vegetable oils. Although trans-isomers of C₁₈ carbon length predominate in partially hydrogenated vegetable oils, trans-isomers of C₂₀ and C₂₂ carbon length occur in partially hydrogenated fish oil. We report a simple silver ion chromatographic combined with capillary GC technique for quantitative analysis of trans-monoenes derived from partially hydrogenated fish oil. Silver nitrate thinlayer chromatographic (TLC) plates are developed in toluene/hexane (50∶50, vol/vol). Fatty acid methyl esters are separated into saturates (R _f 0.79), trans-monoenes (R _f 0.49), cis-monoenes (R _f, 0.27), dienes (R _f, 0.10), and polyunsaturated fatty acids with three or more double bonds remaining at the origin. The isolated trans-monoenes are quantitatively analyzed by capillary GC. The technique of argentation TLC with GC analysis of isolated methyl esters is highly reproducible with 4.8% variation (i.e., coefficient of variation, CV%) in R _f values and 4.3 and 6.9% CV% in quantification within batch and between batch, respectively. Furthermore, the combined technique revealed that direct GC analysis underestimated the trans-content of margarines by at least 30%. In this study, C₂₀ and C₂₂ trans-monoenes were found in relatively large quantities; 13.9% (range 10.3–19.6%) and 7.5% (range 5.3–11.5%), respectively, in margarine purchased in 1995, but these C₂₀ and C₂₂ trans-monoenes were much reduced (0.1%) in a fresh selection of margarine purchased in 1998. Compositional data from labels underestimated the trans-content of margarines, especially those dervied from hydrogenated marine oil. Low levels of C₂₀ trans-monoenes (range 0.1–0.3%) and C₂₂ trans-monoenes (range 0.0–0.1%) were identified in adipose tissue obtained from healthy volunteers in 1995, presumably indicating consumption of partially hydrogenated fish oil.     

Glyceride structure ofCardamine impatiens L. Seed oil

A group of unusual triglycerides, in which one of the acyl groups is a vicinal dihydroxy acid with one of the hydroxyl groups acetylated, has been isolated fromCardamine impatiens L. (Cruciferae) seed oil. Hydrolysis of these triglycerides with castor bean lipase facilitated isolation and identification of a mixture of C₁₈, C₂₀, C₂₂, and C₂₄ hydroxy acetoxy fatty acids. Pancreatic lipase hydrolysis data revealed that these monoacetylated dihydroxy acid residues are esterified exclusively with one of the α-positions of the glycerol moiety. The remaining acyl groups are comprised of ordinary C₁₈ unsaturated acids (which occupy 98% of the β-position), palmitic acid, and C₂₀, C₂₂, and C₂₄ monoenoic fatty acids. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Molecular species of mycolic acid subclasses in eight strains ofMycobacterium smegmatis

Thin layer chromatographic and gas chromatographic separation and mass spectrometric identification of mycolic acid subclasses and molecular species from eight strains ofMycobacterium smegmatis were established. Two major adjacent spots and a lower minor one were detected on silica gel thin layer chromatograms of methyl esters. The most abundant subclass showing the highest R_f value on TLC was that of α-mycolic acids (M₁), the second was that of α′-mycolic acids (M₁′) a shorter homologue than α-mycolates, and the third was the hydroxy mycolic acids (M₄) derived from epoxy mycolic acids. They were identified by gas chromatography-mass spectrometry as their trimethylsilylether derivatives. α′-Mycolic acids were monoenoic acids ranging from C₆₀ to C₆₆ and possessing an α-unit of C_24∶0. Such profiles of α′-mycolic acids were common in eight strains. α-Mycolates were dienoic acids ranging from C₇₅ to C₇₉ and possessing an α-unit of C_24∶0. In most strains, the major molecular species of α-mycolates were odd-carbon-numbered, centering at C₇₇ and C₇₉, possessing a methyl branch in the even-carbon-numbered straight chain. The average carbon number of α-mycolates, from seven strains examined, was about 78, but that of the Takeo strain was 76.3. The profiles of epoxy mycolic acid molecular species composition from eight strains ranging from C₇₅ to C₈₁ were very similar to their M₁ subclass profiles.     

Lipids from the paracloacal glands of the American alligator (Alligator mississippiensis)

Lipids from the paracloacal glands of adult and of immature American alligators (Alligator mississippiensis) were analyzed by gas chromatography-mass spectrometry. Acetate esters (C₁₂−C₁₈) were indicated in the adults' secretions. Immature alligators contain C₁₀−C₁₈ acetates, C₁₂−C₁₈ dodecanoates, tetradecanoates and other high molecular weight esters, and C₁₀−C₁₆ 3-methyl-butanoates. Cholesterol, C₁₆ and C₁₈ free fatty acids, and α-tocopherol (vitamin E) were detected in some samples. The results are compared with those published for South American caiman species.     

Utilization of acid oils in making valuable fatty products by microbial lipase technology

Microbial lipase-catalyzed hydrolysis, esterification, and alcoholysis reactions were carried out on acid oils of commerce such as coconut, soybean, mustard, sunflower, and rice bran for the purpose of making fatty acids and various monohydric alcohol esters of fatty acids of the acid oils. Neutral glycerides of the acid oils were hydrolyzed byCanadida cylindracea lipase almost completely within 48 h. Acid oils were converted into fatty acid esters of short- and long-chain alcohols like C₄, C₈, C₁₀, C₁₂, C₁₆, and C₁₈ in high yields by simultaneous esterification and alcoholysis reactions withMucor miehei lipase as catalyst. Acid oils of commerce can be utilized as raw materials in making fatty acids and fatty acid esters using lipase-catalyzed methodologies.