Analysis of the conjugated trienoic acid containing oil fromFevillea trilobata by13C nuclear magnetic resonance spectroscopy

Oil ofFevillea trilobata has the composition palmitic acid 31%, stearic acid 12%, oleic acid 11%, linoleic acid 7%,cis,trans,cis 9,11,13-octadecatrienoic acid 30% andcis,trans,trans-9,11,13-octadecatrienoic acid 9%. The oil was analyzed and components identified by¹³C NMR spectroscopy; the composition was also confirmed by conventional methods. Assignment of¹³C NMR signals of conjugated trienoic acids is discussed and it is shown that mixtures of isomeric conjugated acids can be analyzed by this method. NRCC No. 17797.     

Occurrence of mixtures of geometrical isomers of conjugated octadecatrienoic acids in some seed oils: Analysis by open-tubular gas liquid chromatography and high performance liquid chromatography

Analytical methods to obtain the detailed compositions of the fatty acids in oils containing more than one conjugated octadecatrienoic acid by open-tubular gas liquid chromatography (GLC) and by reversed-phase high performance liquid chromatography (HPLC) were established. Effective GLC separations ofcis,trans,trans-9,11,13-octadecatrienoic acid (ctt-9,11,13–18∶3),ctc-9,11,13–18∶3,ttc-9,11,13–18∶3,ttt-9,11,13–18∶3,ttc-8,10,12–18∶3, andttt-8,10,12–18∶3 were obtained with an opentubular column coated with the nonpolar liquid phase OV-1 using an instrument having all-glass carrier gas pathways. The HPLC method also gave satisfactory separations for the isomeric conjugated octadecatrienoates on the basis of number of thecis andtrans double bonds. Two or three minor conjugated trienoic acids were found along with the principal conjugated trienoic acid in tung oil, and seed oils of cherry,Prunus sp., Momordica charantia, Trichosanthes anguina, Punica granatum, Catalpa ovata, andCalendula officinalis. The mechanism for the formation of the conjugated trienoic acid mixtures in the seed oils is discussed. TheC. ovata seed oil also containedct andtt-9,12-octadecadienoic acids. Thett isomer is presumed to be a precursor ofttc-9,11,13–18∶3, the main conjugated trienoic acid in this oil.     

OxygenatedTrans-3-olefinic acids in aStenachaenium seed oil

Interesting differences were found in oils from two samples ofStenachaenium macrocephalum (Compositae) seed with dissimilar storage histories. One contained significant amounts of epoxy acids (6.5%) and hydroxy conjugated dienoic acids (5.6%), but the other contained no more than 1% of these oxygenated acids. Characterization of components in the former oil established that the principal epoxy acid (4.0%) is the previously unknowncis-9, 10-epoxy-trans-3,cis-12-octadecadienoic acid. The conjugated dienols include two additional new acids with Δ3 unsaturation (2.5%): 9-hydroxy-trans-3,-trans-10,cis-12-octadecatrienoic and 13-hydroxy-trans-3,cis-9,trans-11-octadecatrienoic acoids. The nonoxygenated acids, except for the large amount (40%) oftrans-3,cis-9,cis-12-octadecatrienoic, are those that commonly occur in seed oils. Presented at the AOCS Meeting, San Francisco, April 1969. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Therans-3-enoic acids ofAster alpinus andArctium minus seed oils

Thetrans-3-enoic acids ofAster alpinus (dwarf aster, rock aster) andArctium minus (burdock) seed oils have been isolated and characterized.Arctium seed oil containstrans-3,cis-9,cis-12-octadecatrienoic acid (9.9%), andAster oil containstrans-3-hexadecenoic (7.1%),rans-3-octadecenoic (1.9%),trans-3,cis-9-octadecadienoic (3.0%),a ndtrans-3,cis-9,cis-12-octadecatrienoic (13.7%) acids.Aster oil also has an epoxy acid as a minor constituent (ca. 2.0%), which has been identified ascis-9,10-epoxy-cis-12-octadecenoic acid.     

Identity and configuration of conjugated fatty acids in certain seed oils

Punicic acid was found in the seed oils ofCayaponia grandifolia, Trichosanthes cordata andT. cucumerina. α-Eleostearic acid was found inMomordica cochinchinensis, M. cochinchinesis varietymixta andM. cymbalaria. The identity of the conjugated triene acid ofAleurites trisperma, Garcia nutans andCyclandrophora laurina was confirmed as α-eleostearic acid (cis, trans, trans configuration). The configuration of kamlolenic acid was proved to becis-9,trans-11,trans-13. The oils ofAleurites remyi andLicania platypus did not contain any conjugated acid. Issued as N.R.C. No. 10589.     

Alkali isomerization of linoleate isomers: Characterization of products

Geometrical isomers of methyl linoleate were reacted with alakli, and the resulting conjugated isomers were separated intotrans,trans;cis,trans; andcis,cis fractions. The position of double bonds in the various fractions was determined by reductive ozonolysis.trans-9,trans-12-Isomer of linoleate formedtrans,trans- andcis,trans-conjugated dienes, whereascis-9,trans-12- andtrans-9,cis-12-isomers in addition formedcis,cis-conjugated dienes. The formation of the products is in accordance with the theoretical predictions. During conjugationtrans double bonds shifted to form atrans bond preferentially. During conjugation ofcis-9,trans-12- andtrans-9,cis-12-linoleate isomers, thecis double bond shifted preferentially over thetrans double bond. A small amount of diene not conjugated was probably a geometrical and positional isomer of the starting material.     

Selective homogeneous hydrogenation of triunsaturated fats catalyzed by tricarbonyl chromium complexes

The need for a selective catalyst to hydrogenate linolenate in soybean oil has prompted our continuing study of various model triunsaturated fats. Hydrogenation of methylβ-eleostearate (methyltrans,trans,trans-9,11,13-octadecatrienoate) with Cr(CO)₃ complexes yielded diene products expected from 1,4-addition (trans-9,cis-12- andcis-10,trans-13-octadecadienoates). Withα-eleostearate (cis,trans,trans-9,11,13-octadecatrienoate), stereoselective 1,4-reduction of thetrans,trans-diene portion yielded linoleate (cis,cis-9,12-octadecadienoate). However,cis,trans-1,4-dienes were also formed from the apparent isomerization ofα- toβ-eleostearate. Hydrogenation of methyl linolenate (methylcis,cis,cis-9,12,15-octadecatrienoate) produced a mixture of isomeric dienes and monoenes attributed to conjugation occurring as an intermediate step. The hydrogenation ofα-eleostearin in tung oil was more stereoselective in forming thecis,cis-diene than the corresponding methyl ester. Hydrogenation of linseed oil yielded a mixture of dienes and monoenes containing 7%trans unsaturation. We have suggested how the mechanism of stereoselective hydrogenation with Cr(CO)₃ catalysts can be applied to the problem of selective hydrogenation of linolenate in soybean oil. No. Market. Nutr. Res. Div., ARS, USDA.     

Hydrogenation of linolenate. VII. Separation and identification of isomeric dienes and monoenes

Isomeric dienes and monoenes produced by partial hydrogenation of linolenic acid have been separated by the combined use of low-temp crystallization and countercurrent distribution.Cis, trans dienes have been separated fromcis, cis dienes.Cis, cis conjugatable dienes have been partially separated fromcis,cis nonconjugatable dienes. Dienes with onetrans double bond were separated by gas chromatography into two groups:cis, trans andtrans, cis. Individual positional isomers could not be separated. Cis-9 monoene was separated fromcis-12,cis-15, andtrans monoenes by low-temp crystallization. Countercurrent distribution at 3,000 transfers only partially separated this mixture ofcis-12,cis-15, andtrans monoenes. The double bond in bothcis andtrans monoenes was found in all carbon positions, 7 through 16, showing for the first time that the 15, 16 bond of linolenic acid had moved away from the carboxyl. The majorcis bonds remained at carbons 9, 12, and 15. Combination of countercurrent distribution fractions has produced samples containing 95%cis, cis dienes; 90%cis, trans ortrans, cis dienes; 95%cis monoenes; and 90%trans monoenes. Presented at the AOCS meeting at Chicago, Ill., 1961. A laboratory of the No. Utiliz. Res. & Dev. Div., ARS, U.S.D.A.     

Kamlolenic acid and other conjugated fatty acids in certain seed oils

The major fatty acid of the seed oil ofTrewia nudiflora is shown to be α-kamlolenic acid, not α-eleostearic acid as believed earlier. Other conjugated acids were found and identified in seed oils not previously studied, viz., α-eleostearic acid inParinari insularum andRicinodendron rautanenii; trans,8-trans,10-cis,12-octadecatrienoic acid inCalendula stellata. The identity of the conjugated acids in four other seed oils was established, viz., α-eleostearic acid inPrunus yedoensis andValeriana officinalis; punicic acid inCucurbita digitata andC. palmata. Issued as N.R.C. No. 9063.     

Partial reduction of α-Eleostearic acid with hydrazine

The following products are formed during partial reduction of α-eleostearic acid with hydrazine:cis,trans-9,11-octadecadienoic andtrans,trans-11,13-octadecadienoic acids;cis-9-,trans-11- andtrans-13-octadecenoic acids; and stearic acid. The double bonds are reduced individually in the conjugated triene and also in the conjugated dienes that are formed. However, the reduction is selective since thetrans-11 double bonds in the conjugated triene is reduced only slightly to yield the isolated 9,13-diene. Thetrans double bond of thecis,trans conjugated diene reduces at a faster rate than thecis bond. No differences were observed in the rate of reduction of thecis-9 andtrans-13 bonds in the triene or of the bonds in thetrans,trans conjugated diene. No. Utiliz. Res. & Dev. Div., ARS, USDA.     

Teucrium depressum seed oil: A new source of fatty acids with Δ--unsaturation

The seed oil ofTeucrium depressum Small yields two unusual trienoid components, all-cis-5,9,12-octadecatrienoic acid (6.7%) andtrans-5,cis-9,cis-12-octadecatrienoic acid (2.0%). A third unusual component, identified ascis-5,cis-9-octadecadienoic acid, also occurs in this oil as a trace constituent. Student trainee, 1965–1968. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Formation of conjugated diene and triene products in lipoxygenase oxidation of C18, C20, C22 PUFAs

The determination of conjugated diene formation revealed that the mol % conversions of allcis-6,9,12-octadecatrienoic acid [γ-linolenic, 18:3(n-6)], allcis-5,8,11,14-eicosatetraenoic acid [arachidonic, 20:4(n-6)], allcis-5,8,11,14,17-eicosapentaenoic acid [20:5(n-3)], and allcis-4,7,10,13,16,19-docosahexaenoic acid [22:6(n-3)] into conjugated diene products by soybean lipoxygenase-1 at pH 9.0 were 84, 86, 60 and 40% of that of allcis-9,12-octadecadienoic acid [linoleic, 18:2(n-6)], respectively. On the other hand, the conversions of allcis-9,12,15-octadecatrienoic acid [α-linolenic, 18:3(n-3)], allcis-5,9,12-octadecatrienoic acid (c5,c9,c12-18:3), andtrans-5,cis-9,cis-12-octadecatrienoic acid (t5,c9,c12-18:3) were equal to that of 18:2(n-6). The lowering of the conjugated diene formation in the oxidation of 18:3(n-6), 20:4(n-6), 20:5(n-3), and 22:6(n-3) by the lipoxygenase was thought to be caused by the further oxidation of conjugated diene monohydroperoxides to yield conjugated triene products. For this reason, the conventional lipoxygenase method gave erroneous values forcis,cis-methylene interrupted polyunsaturated fatty acids (PUFA) in oils containing a large amount of 20:5(n-3) and 22:6(n-3) such as fish oils. However, by changing the pH of reaction mixtures from 9.0 to 11.0, the secondary oxidation of conjugated diene monohydroperoxides was completely inhibited, and the PUFA values in fish oils obtained by this improved method were in good agreement with those obtained by a GLC method.     

The geometric isomers of conjugated octadecadienoates from dehydrated methyl ricinoleate

The dehydration of methyl ricinoleate by heatingin vacuo in the presence of KHSO₄ resulted in the formation of the following conjugated octadecadienoates expressed as a percentage of the final product:cis, trans (trans, cis), 14.3;cis, cis, 11.2;trans, trans, 7.3. The isomers contained the double bonds predominantly in the 9,11 position but the possible presence of traces of 8,10 and other conjugated isomers is not excluded. Using urea “inclusion” fractionation and low temp crystallization from acetone methyl,cis-9,cis-11-octadecadienoate was isolated. The methyl esters of commercially dehydrated castor oil fatty acids on the other hand, contained the following percentages of conjugated octadecadienoate isomers:cis, trans (trans, cis), 20.3;cis, cis, 8.0;trans, trans, 5.4. From these mixtures conc ofcis, trans (trans, cis)- andtrans, trans-octadecadienoates were prepared by fractional distillation and low temp crystallization. It was found that the conjugated octadecadienoates consisted of mixtures of positional isomers with double bonds mainly in the 8,10 and 9,11 positions with lesser amounts in the 7,9 and 10,12 positions.     

Essential fatty acid-deficient rats: II. On the fatty acid composition of partially hydrogenated arachis,soybean and herring oils and of normal,refined arachis oil

The fatty acid composition of partially hydrogenated arachis (HAO), partially hydrogenated soybean (HSO) and partially hydrogenated herring (HHO) oils and of a normal, refined arachis oil (AO) was studied in detail by means of direct gas liquid chromatography, ultraviolet and infrared spectrophotometry and by thin layer chromatography fractionation on silver nitrate-silica gel plates followed by gas liquid chromatography. It was shown that the partially hydrogenated oils all contained fatty acids withtrans double bonds. In the plant oils, thetrans acids were present mainly as elaidic acid. The HHO showed an almost equal distribution betweentrans 18∶1 ω9,trans 20∶1 ω>9 andtrans 22∶1 ω>9. Sometrans configuration was also found in the C₂₀-and C₂₂-dienes and trienes of the HHO. In all the oils, conjugated fatty acids were present in minor amounts only (<0.5%). Special attention was given to the ω-acids known to be of specific nutritional value. The HSO contained about 32% linoleic acid, whereas the content ofcis, trans+trans, cis andtrans, trans octadecadienoic isomers was 1.7% and 0.5%, respectively. The amount of linoleic acid in the HSO was even higher than that of AO (29%). The HAO contained only 0.8% 18∶2 ω6 (linoleic acid). Further, two 18∶2 fatty acids with ω>6, acis, cis and atrans, trans isomer, were present in small amounts. The HHO contained 0.5% 18∶2 ω6 (linoleic acid). Isomers of 18∶2 ω>6 were also found in the HHO. They may be hydrogenation products of higher unsaturated C₁₈-acids orginally present. All the C₂₀- and C₂₂-dienes and trienes were shown to have an ω-chain greater than 6. Fatty acids with ω6-structure were not formed during partial hydrogenation of the oils studied.     

Methyl punicate,Alpha andBeta eleostearate.Cis,trans isomerism and structure

Methyl esters ofbeta eleostearic,alpha eleostearic, and punicic acids have been isomerized with iodine and light to the same equilibrium mixture of 64%beta, 33%alpha, and 2.6% punicate structures. The course of isomerization is in agreement with the following structures:trans, trans, trans forbeta; cis, trans, trans foralpha; andcis, trans, cis for punicate, in agreement with structures provenvia synthesis by Crombie and Jackson. There is some theoretical and experimental evidence that the center double bond of this type of conjugated triene isomerizes less readily than the outer double bonds. Paper No. 253, Journal Series, General Mills Inc., Central Research.     

Analyses of fatty acid isomers in two commercially hydrogenated soybean oils

A conventional shortening and a hydrogenated winterized oil have been investigated to determine their composition of natural and isomeric fatty acids. Two solvent systems were applied in countercurrent distributions: the acetonitrile pentane-hexane system for separation of monoenoates from dienoates and the methanolic silver nitrate pentane-hexane system for separation of geometric isomers. Whilecis andtrans monoenoates were well resolved, the separation ofcis,cis fromcis,trans dienoates was complicated by the presence of positional isomers. The fractions isolated were oxidatively cleaved, and the esters of the resultant acids were quantitatively analyzed by gas-liquid chromatography. Although the amounts of saturated components of the two fat products were similar, the percentage oftrans isomers of the shortening was more than twice that of the winterized oil. The amount of oleic acid (cis-9-octadecenoic) was 19.6% for the shortening and 25.4% for the winterized oil. The shortening contained 13.3% linoleic acid (cis,cis-9,12-octadecadienoic), whereas the winterized oil contained 30% linoleic acid. Although our primary interest was in the estimation ofcis-9-octadecenoic andcis,cis-9,12-octadecadienoic acids, the completeness of cleavage data makes it possible to estimate all geometric and positional monoenoate and dienoate isomers in the two fat products. Presented at AOCS meeting in New Orleans, La., May, 1962. A laboratory of the No. Utiliz. Res. and Dev. Div., ARS, USDA.     

Linoleic acid isomers in heat treated sunflower oils

Heat treatment of sunflower oil resulted in the formation of linoleic geometrical and positional isomers. These isomers were isolated using a combination of column chromatography, urea fractionation, high performance liquid chromatography (HPLC) on a C18 reverse phase column and silver nitrate thin layer chromatography (TLC). Each component was submitted to hydrazine reduction and the resulting monoenes to AgNO₃-TLC. The resultingcis andtrans fractions were submitted to ozonolysis in BF₃-MeOH in order to determine the position of the ethylenic bonds. The major isomers were thecis, trans andtrans, cis 18∶2 Δ9, 12, thetrans, trans 18∶2 Δ9, 12 and somecis, trans, trans, cis andtrans, trans 18∶2 conjugated dienes. Thecis, trans andtrans, cis conjugated dienes were the Δ9, 11, Δ10, 12, Δ11, 13 and Δ12, 14 while thetrans, trans isomers were the Δ9, 11, Δ10, 12 and Δ11, 13. These C18∶2 isomers also were detected in oils collected from restaurants and market vendors. Presented in part at the AOCS/JOCS annual meeting in Honolulu, Hawaii, in May 1986.     

Hydrolysis of linoleate geometric isomers byGeotrichum candidum lipase

Lipase (EC 3.1.1.3) from the microorganismGeotrichum candidum preferentially hydrolyzescis-9 18∶1 andcis,cis-9,12 18∶2 from triacylglycerols, largely ignoring all other positional isomers ofcis 18∶1 as well astrans-9 18∶1. To obtain additional information about the specificity of the enzyme, two triacylglycerols were prepared and utilized as substrates. The lipase hydrolyzed 85%cis,cis-9,12 18∶2 and 15%trans,trans-9,12 18∶2 from the triacylglycerol, containing ca. 50% of each acid. From the triacylglycerol containing 46.3%cis,trans-9,12 18∶2 and 53.7%trans,cis-9,12 18∶2, 44.8 and 55.2% of the two acids were hydrolyzed. Therefore the enzyme discriminated against thetrans,trans isomer but not between thecis,trans andtrans,cis isomers. Scientific contribution No. 535, Agricultural Experiment Station, University of Connecticut. ARS, USDA.     

TheTrans-6 fatty acids ofPicramnia sellowii seed oil

The C₁₈ monoenoic acids inPicramnia sellowii Planch. seed oil include bothcis-andtrans-6-octadecenoic acids, as well as oleic acid. The hexadecenoic acids are also thecis- andtrans-Δ6-isomers, and the eicosenoic acids have Δ6-unsaturation of undetermined geometric configuration. The C₁₈ polyenoic acids detected are 9,12- and 6,9-octadecadienoic and 9,12,15- and 6,9,12-octadecatrienoic acids. Partial investigation of another species,P. pentandra Sw., revealed its oil to have a similar fatty acid composition. Presented in part at AOCS Meeting, New York, October 1968. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Composition and structure of some consumer-available edible fats

Samples of polyunsaturated margarines, table margarines, hard cube polyunsaturated vegetable oil, hard cube vegetable oil, animal fat, and blends of vegetable oil and animal fat were analyzed for fatty acid composition, percentage ofcis,cis-methylene interrupted polyunsaturated fatty acids, percentage isolatedtrans-unsaturation, and percentage conjugated diene. Gas liquid chromatography was used to separate and measure the geometric isomers of the octadecaenoic and octadecadienoic acids. Selected samples were analyzed for the content of positional isomers in theircis monoene andtrans monoene fractions, and for the percentage of fatty acid esterified in the 2-position of their triglycerides.