trans-Polyunsaturated fatty acids in French edible rapeseed and soybean oils

The fatty acid compositions of rapeseed and soybean oils marketed in France have been determined by gas liquid chromatography on a fused-silica capillary column coated with a 100% cyanopropyl polysiloxane stationary phase. Under the operating conditions employed, methyl esters of linolenic acid geometrical isomers could be separated and quantitated easily without any other complementary technique. With only one exception, all samples under study (eight salad oils and five food samples) contain geometrical isomers of linolenic acid in measurable, although variable, amounts. Totaltrans-18:3 acids may account for up to 3% of total fatty acids. This value corresponds to a degree of isomerization (percentage oftrans isomers relative to total octadecatrienoic acids) of 30%. Examination of our data indicates that the distribution pattern of linolenic acid geometrical isomers does not depend on the degree of isomerization. The two main isomers always have thec,c,t and thet,c,c configurations. These isomers occur in the almost invariable relative proportions of 47.8±1.7% and 41.1±1.0%, respectively. The third mono-trans isomer is present in lower amounts−6.5±0.7%. The only di-trans isomer that can be quantitated with sufficient accuracy is thet,c,t isomer (4.9±1.5%). Mono-trans isomers of linoleic acid are also present in these oils. However, their maximum percentages are lower than those determined for linolenic acid geometrical isomers. In the oils showing the highest degrees of isomerization,trans isomers of linoleic acid account for 0.5% (rapeseed oils) and 1% (soybean oils) of total fatty acids. Taking into account all data, it would appear that the probability of isomerization of linolenic acid is about 13–14 times that of linoleic acid.     

Deodorization of vegetable oils: Prediction of trans polyunsaturated fatty acid content

Kinetics of the formation of trans linoleic acid and trans linolenic acid were compared. Pilot plant-scale tests on canola oils were carried out to validate the laboratory-scale kinetic model of geometrical isomerization of polyunsaturated fatty acids described in our earlier publication. The reliability of the model was confirmed by statistical calculations. Formation of the individual trans linoleic and linolenic acids was studied, as well as the effect of the degree of isomerization on the distribution of the trans fatty acid isomers. Oil samples were deodorized at temperatures from 204 to 230°C from 2 to 86 h. Results showed an increase in the relative percentage of isomerized linolenic and linoleic acid with an increase in either the deodorization time or the temperature. The percentage of trans linoleic acid (compared to the total) after deodorization ranged from <1 to nearly 6%, whereas the percentage of trans linolenic acid ranged from <1 to >65%. Applying this model, the researchers determined the conditions required to produce a specially isomerized oil for a nutritional study. The practical applications of these trials are as follows: (i) the trans fatty acid level of refined oils can be predicted for given deodorization conditions, (ii) the conditions to meet increasingly strict consumer demands concerning the trans isomer content can be calculated, and (iii) the deodorizer design can be characterized by the deviation from the theoretical trans fatty acid content of the deodorized oil.     

Preparative Separation of <Emphasis Type="Italic">cis</Emphasis>- and <Emphasis Type="Italic">trans</Emphasis>-Isomers of Unsaturated Fatty Acid Methyl Esters Contained in Edible Oils by Reversed-Phase High-Performance Liquid Chromatography

In order to measure exactly the trans-fatty acids content in food materials, a preparative group separation of cis- and trans-isomers of unsaturated fatty acid methyl esters (FAMEs) was achieved by an isocratic reversed-phase HPLC (RP-HPLC) method. The trans-isomers of 16:1, 18:1, 18:2, 18:3, 20:1 and 22:1 FAMEs were readily separated from the corresponding cis-isomers by a COSMOSIL Cholester C18 column (4.6 mm I.D. × 250 mm, Nacalai Tesque) or a TSKgel ODS-100Z column (4.6 mm I.D. × 250 mm, TOSOH), using acetonitrile as the mobile phase. This method was applied for determining the trans-18:1 fatty acid content in partially hydrogenated rapeseed oil. The methyl esters of cis- and trans-18:1 isomers of the oil were collected as two separate fractions by the developed RP-HPLC method. Each fraction was analyzed by gas chromatography (GC) for both qualitative and quantitative information on its positional isomers. By a combination of RP-HPLC and GC methods, a nearly complete separation of cis- and trans-18:1 positional isomers was achieved and the trans-18:1 fatty acid content was able to be evaluated more precisely than is possible by the direct GC method. The reproducibility of cis- and trans-18:1 isomers fractionated by the RP-HPLC method was better than 98%. These results suggested that the preparative RP-HPLC method developed in this study could be a powerful tool for trans-fatty acid analysis in edible oils and food products as an alternative to silver-ion chromatography.     

Further studies on artificial geometrical isomers of α-linolenic acid in edible linolenic acid-containing oils

Fifteen samples of commercial edible soybean and rapeseed oils (and mixtures of these) from Belgium, Great Britain and Germany have been analyzed for theirtrans-polyunsaturated fatty acid content. Only one sample out of the 13 refined samples, and the two cold-pressed samples, contained trace amounts oftrans isomers. Others contained between 1 and 3.3% of their total fatty acids as geometrical isomers of linoleic and linolenic acids. The degree of isomerization (DI) of linolenic acid varied between 10.5 and 26.9%. Combining results obtained in this study together with corresponding data for French oils (totalling 21 samples) indicates that the relative percentages of individual linolenic acid geometrical isomers depend on linolenic acid DI. Relationships linking these parameters could be approximated by straight lines, at least for DIs lying between 9 and 30%. Extrapolation to DI=0 suggests that the relative probabilities of isomerization of double bonds in positions 9, 12, and 15 are 41.7, 6.1 and 52.1%, respectively, at the very beginning of the isomerization reaction. At that time, the probability of a simultaneous isomerization of double bonds in positions 9 and 15 is close to zero. Thet,c,t isomer is apparently formedvia thec,c,t and thet,c,c isomers, the former being somewhat more prone to a second geometrical isomerization than the latter. The relative proportion of thec,t,c isomer is practically independent from the DI, at least between 9 and 30%, which would suggest that this isomer is an “end-product” of thecis-trans isomerization reaction.     

Positional and geometrical isomers of linoleic acid in partially hydrogenated oils

The geometrical and positional isomers of linoleic acid of a partially hydrogenated canola oil-based spread were isolated and identified. Through partial hydrazine reduction and mass spectral studies,cis-9,trans-13 octadecadienoic acid was identified as the major isomer. Other quantitatively important isomers characterized werecis-9,trans-12;trans-9,cis-12 andcis-9,cis-15. These four were also the major isomers in margarine based on common vegetable oils. A number of minor isomers were detected and some structures identified weretrans-9,trans-12;trans-8,cis-12;trans-8,cis-13;cis-8,cis-13;trans-9,cis-15;trans-10,cis-15 andcis-9,cis-13. The proportions of the various isomers are given for some margarines in the Canadian retail market. The amounts oftrans-9,trans-12 isomer in Canadian margarines were generally below 0.5% of the total fatty acids.     

Fatty acid composition of French infant formulas with emphasis on the content and detailed profile oftrans fatty acids

The aim of the present study was to identify and quantitatetrans isomers of C18 fatty acids in some French infant formulas. Twenty powdered infant formulas were purchased in pharmacies and supermarkets in order to assess theirtrans mono- and poly-unsaturated fatty acids content. The fatty acid profiles were examined using methyl and isopropyl ester derivatives. The combination of gas-liquid chromatography, high-performance liquid chromatography, and silver nitrate thin-layer chromatography was needed to describe the detailed fatty acid compositions of the samples, includingtrans isomers of unsaturated C18 fatty acids. All the samples containedtrans isomers of C18∶1 acid (mean level 1.97±0.28% of total fatty acids), with vaccenic acid being generally the major isomer (15 out of 20 samples), thus indicating the origin from bovine milk. All the formulas also contained various isomers of linoleic and α-linolenic acids, but at lower levels.Trans PUFA isomers are the same as those present in deodorized oils. In conclusion, all the infant formulas analyzed in this study contained sometrans fatty acids, including isomers of essential fatty acids. This should be taken into account in the dietary intake of the newborn.     

Analysis of C18:1 cis and trans fatty acid isomers by the combination of gas-liquid chromatography of 4,4-dimethyloxazoline derivatives and methyl esters

Trans fatty acids in foods are usually analyzed by gas-liquid chromatography (GLC) of fatty acid methyl esters (FAME). However, this method may produce erroneously low values because of insufficient separation between cis and trans isomers. Separation can be optimized by preceding silver-ion thin-layer chromatography (Ag-TLC), but this is laborious. We have developed an efficient method for the separation of 18-carbon trans fatty acid isomers by combining GLC of FAME with GLC of fatty acid 4,4-dimethyloxazoline (DMOX) derivatives. We validated this method against conventional GLC of FAME, with and without preceding Ag-TLC. Fatty acid isomers were identified by comparison with standards, based on retention times and mass spectrometry. Analysis of DMOX derivatives allowed the 13t, 14t, and 15t isomers to be separated from the cis isomers. The combination of the GLC analyses of FAME and DMOX derivatives gave results comparable with those obtained by GLC of FAME after preceding Ag-TLC, while saving about 100 h of manpower per 25 samples. It allowed the identification and quantitation of 11 trans and 8 cis isomers and resulted in 25% higher values for total C_18:1 trans, compared with the analysis of FAME alone. The combination of DMOX and FAME analyses, as applied to the analysis of 14 foods that contained ruminant fat and partially hydrogenated vegetable and fish oils, indicated that the most common isomers were 11t in ruminant fats, 9t in partially hydrogenated fish fats, and either 9t or 10t in partially hydrogenated vegetable fats. The combination of GLC analyses of FAME and DMOX derivatives of fatty acids improves the quantitation of 18-carbon fatty acid isomers and may replace the laborious and time-consuming Ag-TLC.     

Effect of fatty acid positional distribution and triacylglycerol composition on lipid by-products formation during heat treatment: II. Trans isomers

This study examined the effect of the fatty acid positional distribution and of the triacylglycerol (TG) composition on heat-induced trans isomerization of linoleic and linolenic acids. For this, we synthesized diacid TG molecules that were acylated only with linoleic acid (L) or with linolenic acid (Ln) along with palmitic acid (P). The fatty acid of interest was positioned either in the central position (PLP and PLnP, respectively) or in one of the two outer positions (PPL and PPLn, respectively). Monoacid TG, i.e., trilinolein and trilinolenin, were also synthesized and mixed with tripalmitin in a 1:2 ratio. This model TG was also compared to another TG model, which consisted of a canola oil and its randomized counterpart whose fatty acid positional distribution and TG composition were determined by means of high-performance liquid chromatography. After heating, the content of trans isomers was determined by gas-liquid chromatography with a polar capillary column. In model TG, polyunsaturated fatty acids in monoacid TG (LLL and LnLnLn) exhibited the highest degree of isomerization, compared to diacid TG, and this effect was greatest at 220°C. At this temperature, an effect of the TG structure was observed only with linolenic acid. In that situation, 18:3n-3 acylated in the central position of the TG molecule (PLnP) displayed the highest sensitivity to trans geometrical isomerization. Although to a lesser extent, the same trends as for the pure TG model were observed with the canola oil model with regard to the influence of the fatty acid positional distribution and TG molecular species.     

Effect of fatty acid positional distribution and triacylglycerol composition on lipid by-products formation during heat treatment: II. Trans isomers

This study examined the effect of the fatty acid positional distribution and of the triacylglycerol (TG) composition on heat-induced trans isomerization of linoleic and linolenic acids. For this, we synthesized diacid TG molecules that were acylated only with linoleic acid (L) or with linolenic acid (Ln) along with palmitic acid (P). The fatty acid of interest was positioned either in the central position (PLP and PLnP, respectively) or in one of the two outer positions (PPL and PPLn, respectively). Monoacid TG, i.e., trilinolein and trilinolenin, were also synthesized and mixed with tripalmitin in a 1:2 ratio. This model TG was also compared to another TG model, which consisted of a canola oil and its randomized counterpart whose fatty acid positional distribution and TG composition were determined by means of high-performance liquid chromatography. After heating, the content of trans isomers was determined by gas-liquid chromatography with a polar capillary column. In model TG, polyunsaturated fatty acids in monoacid TG (LLL and LnLnLn) exhibited the highest degree of isomerization, compared to diacid TG, and this effect was greatest at 220°C. At this temperature, an effect of the TG structure was observed only with linolenic acid. In that situation, 18:3n-3 acylated in the central position of the TG molecule (PLnP) displayed the highest sensitivity to trans geometrical isomerization. Although to a lesser extent, the same trends as for the pure TG model were observed with the canola oil model with regard to the influence of the fatty acid positional distribution and TG molecular species.     

Deodorization of vegetable oils. Part I: Modelling the geometrical isomerization of polyunsaturated fatty acids

Laboratory-scale treatments of canola oils similar to deodorization were carried out by applying the following conditions: reduced pressure with nitrogen or steam stripping at different temperatures ranging from 210 to 270°C for 2–65 h. The formation of the group of trans linolenic acid isomers follows a first-order reaction and the kinetic constant varies according to the Arrhenius’ law. Similar results were observed for the trans isomerization of linoleic acid. Based on these experiments, a mathematical model was developed to describe the isomerization reaction steps occurring in linoleic and linolenic acids during deodorization. The calculated degrees of isomerization are independent of the composition of the oil but related to both time and temperature of deodorization. The degree of isomerization of linolenic acid is unaffected by the decrease of this acid content observed during the deodorization. Deodorization at about 220–230°C appears to be a critical limit beyond which the linolenic isomerization increases very strongly. The newly established model can be a tool for manufacturers to reduce the total trans isomer content of refined oils, and was applied to produce a special selectively isomerized oil for a European Nutritional Project.     

Characterization of γ-linolenic acid geometrical isomers in borage oil subjected to heat treatments (deodorization)

Heating of borage oil, either under vacuum as a model or during steam-vacuum deodorization, produces artifacts that are geometrical isomers of γ-linolenic acid (cis-6,cis-9,cis-12 18∶3 acid). In a first approach, we have studied the behavior of these fatty acids in the form of either methyl or isopropyl esters on two capillary columns (CP-Sil 88 and DB-Wax). From this study, it appears that the DB-Wax capillary column is the best suited analytical tool to study in some detail γ-linolenic acid geometrical isomers. In a second approach, the structure of these isomers was formally established by combining several analytical techniques: Argentation thin-layer chromatography, comparison of the equivalent chainlengths with those of isomers present in NO₂-isomerized borage oil on two different capillary columns, partial hydrazine reduction, oxidative ozonolysis, gas chromatography coupled with mass spectrometry and gas chromatography coupled with Fourier transform infrared spectroscopy. The two main isomers that accumulate upon heat treatments are thetrans-6,cis-9,cis-12 andcis-6,cis-9,trans-12 18∶3 acids with minor amounts ofcis-6,trans-9,cis-12 18∶3 acid. One di-trans isomer, supposed to be thetrans-6,cis-9,trans-12 18∶3 acid, is present in low although noticeable amounts in some of the heated oils. The content of these artificial fatty acids increases with increasing temperatures and duration of heating. The degree of isomerization (DI) of γ-linolenic acid is less than 1% when the oil is deodorized at 200°C for 2 h. Heating at 260°C for 5 h increases the DI up to 74%. Isomerization of γ-linolenic acid resembles that of α-linolenic (cis-9,cis-12,cis-15 18∶3) acid in several aspects: The same kinds and numbers of isomers are formed, and similar degrees of isomerization are reached when the octadecatrienoic acids are heated under identical conditions. It seems that the reactivity of a double-bondvis-à-vis cis-trans isomerization is linked to its relative position, central or external, and not to its absolute position (Δ6, 9, 12 or 15).     

Trans-18:1 acid content and profile in human milk lipids. Critical survey of data in connection with analytical methods

This study presents an in-depth, critical survey of the current knowledge about trans- 18:1 acid content and profile in human milk lipids. Emphasis is placed on the analytical methods employed to quantitate trans- 18:1 acids, most of which lead to imprecise quantitative data. It is demonstrated that data obtained by single gas-liquid chromatography (GLC) on polar capillary columns are underestimates by 25–40%. Several experiments indicate that the total content of trans-18:1 acids in human milk is directly related to the quantities ingested the previous day(s), provided no gross weight loss occurs during breast-milk feeding. Equations have been proposed to describe this relationship, and apparently the percentage of trans-18:1 isomers, relative to total fatty acids, is approximately three-fourths the quantity (in g) ingested by lactating mothers. That is, the determination of the trans-18:1 acid percentage in human milk is a convenient means to estimate trans-18:1 acid consumption by corresponding populations. Adapted methods (i.e., silver-ion thin-layer chromatography, coupled with GLC on long polar capillary columns) allow accurate quantitation of most individual trans- 18:1 acids, more particularly of the trans-Δ16 isomer. This determination, along with a knowledge of the distribution of individual isomers in ruminant fats and partially hydrogenated oils, is a convenient means to estimate the relative contribution of these two dietary sources to the distribution of individual trans-18:1 isomers in human milk lipids. A comparison of human milk and infant formulas is made with regard to trans-18:1 acid content and profile. Important differences are noted between data from European countries and from North America.     

Trans-18:1 acids in french tub margarines and shortenings: Recent trends

The fatty acid composition of twelve French tub margarines and three industrial shortenings was established with particular attention to theirtrans-18:1 acid content. Four of the twelve margarines (including two major brands, with 60% of market share) were devoid oftrans isomers, one contained less than 2%trans-18:1 acids, whereas the seven others had a mean content of 13.5 ± 3.6%trans isomers. Four years ago, no margarines with 0%trans-18:1 acids could be found. It is deduced that the recent Dutch and American studies on possible effects oftrans acids on human health (serum cholesterol, heart disease risks) may have had some influence on French margarine manufacturers. Presently, an average French tub margarine contains only 3.8% oftrans-18:1 acids instead of 13% four years ago. To protect brand names, some manufacturers have replaced partially hydrogenated oils with tropical fats or fully hydrogenated oils. On the other hand, two of the three shortenings had high levels oftrans-18:1 acids: 53.5 and 62.5%. This last value, obtained for a sample of hydrogenated arachis oil, seems to be one of the highest values ever reported for edible hydrogenated oils. In this sample,trans-18:1 plus saturated acids accounted for 85% of total fatty acids. This would indicate that shortening producers and users are not yet aware of recent dietary recommendations, probably because these products are not easily identifiable by consumers in food items, in contrast to margarines.     

13C nuclear magnetic resonance spectral confirmation of Δ6-and Δ7-trans-18:1 fatty acid methyl ester positional isomers

Trans octadecenoic acid methyl ester isomers were obtained from a partially hydrognated soybean oil and isolated by silver-ion high-performance liquid chromatography. Recently, the double-bond positions for nine individual trans octadecenoic acid positional isomers (Δ8 through Δ16) were confirmed by gas chromatography-electron ionization mass spectrometry after derivatization to 2-alkenyl-4,4-dimethyloxazoline. In this communication, the presence of two additional trans-18:1 fatty acid methyl ester positional isomers (Δ6 and Δ7) in the same mixture is confirmed by ¹³C nuclear magnetic resonance spectroscopy. The identity of the Δ5-trans-18:1 fatty acid methyl ester positional isomer is inferred. Summer student researcher.     

Cis-trans isomerization of octadecatrienoic acids during heating. Study of pinolenic (cis-5,cis-9,cis-12 18∶3) acid geometrical isomers in heated pine seed oil

To understand the heat-inducedcis-trans isomerization of ethylenic bonds in octadecatrienoic acids, pine seed oil, which contains the unusual nonmethylene-interrupted pinolenic (cis-5,cis-9,cis-12 18∶3) acid as a major component, was heated under vacuum at 240°C for 6 h together with linseed and borage oils. As a results, a small percentage of pinolenic acid undergoescis-trans isomerization. The main isomer that accumulates is thetrans-5,cis-9,trans-12 18∶3 acid. Minor amounts of the three mono-trans isomers are also present. Identification of isomers was realized by combining gas-liquid chromatography on a CP Sil 88 capillary column, argentation thin-layer chromatography and comparing the equivalent chainlengths of artifacts to those of isomers present in NO₂-isomerized pine seed oil. Hydrazine reduction was used to demonstrate that there was no positional shift of double bonds. Heat-induced geometrical isomerization of pinolenic acid differs from that of α- and γ-linolenic acids in at least two aspects. The reaction rate is slower (about one-fourth), and mono-trans isomers are formed in low amounts.     

Combining Results of Two GC Separations Partly Achieves Determination of All <Emphasis Type="Italic">cis</Emphasis> and <Emphasis Type="Italic">trans</Emphasis> 16:1, 18:1, 18:2 and 18:3 Except CLA Isomers of Milk Fat as Demonstrated Using Ag-Ion SPE Fractionation

Milk fat is a complex mixture of geometric and positional isomers of monounsaturated and polyunsaturated, including short-, long- and branch-chain fatty acids (FAs). There has been partial success to resolve this mixture of FAs using different GC temperature programs, or a combination of GC isothermal and temperature programs. To overcome the problem associated with overlapping isomers prior silver-ion separation was recommended. However, this procedure is time consuming and not practical for routine analysis. In addition, previous methods focused mainly on the trans and cis isomers of 18:1. The present method takes advantage of differences in the relative elution times between different types of FAs. The method involved analyzing each milk fat using the same highly polar 100-m capillary column and GC instrument, and conducting two separations using temperature programs that plateau at 175 and 150 °C. The relative shift among the geometric and positional isomers at these two temperature settings was enough to permit identification of most of the trans and cis 16:1, 18:1 and 20:1, the c/t-18:2 and the c/c/t-18:3 isomers found in milk fat. The identity of these FAs was confirmed by prior separation of the total fatty acid methyl esters (FAMEs) of milk fat using Ag⁺-SPE columns, and comparing the fractions to the total milk fat. The Ag⁺-SPE technique was modified to obtain pure saturated, trans- and cis-monounsaturated and diunsaturated FAMEs. By combining the results from these two separate GC analyses, knowing the elution order, it was possible to determine most of the geometric and positional isomers of 16:1, 18:1, 20:1, 18:2 and 18:3 without a prior silver-ion separation. Only few minor FAs could not be resolved, notable the conjugated linoleic acid isomers that still required the complimentary Ag⁺-HPLC separation. The two GC temperature programs have been successfully used to routinely analyze most FA isomers in total milk and beef fats in about 200 min without the use of prior silver-ion separations.     

Gas-liquid chromatographic analysis of geometrical and positional octadecenoic and octadecadienoic acid isomers produced by catalytic hydrogenation of linoleic acid on Ir/Al2O3

Catalytic hydrogenation of linoleic acid was studied on Ir/Al₂O₃. A detailed analysis of geometrical and positional isomers of octadecenoic acid (18:1) in the products was performed by capillary gas-liquid chromatography with a new capillary column coated with isocyanopropyl trisilphenylene siloxane (TC-70). Well-resolved peaks of 10 species of 18:1 were observed in the product. In addition to monoenoic acid isomers, four species of trans-dienoic isomers and conjugated dienoic isomers were found. From the distribution of 18:1 isomers, it was found that the double bond closer to the methyl end (Δ12) showed higher reactivity than that closer to the carboxyl end (Δ9) for hydrogenation. Because cis-8 18:1 and trans-8 18:1 were not observed but cis-10 18:1 and trans-10 18:1 were observed in the products, the double-bond Δ9 did not migrate to the carboxyl end but migrated to the methyl end. On the other hand, the Δ12 bond migrated to both methyl and carboxyl ends. From the distribution of 18:1 isomers in the reaction pathway, the hydrogenation of linoleic acid proceeds via half-hydrogenation states. Cis-18:1 isomers were produced predominantly in the initial stage of the reaction, while trans-18:1 isomers were produced during progress of the reaction. The cis/trans and positional isomerization took place by readsorption of 18:1 produced by the partial hydrogenation of linoleic acid.     

Analysis oftrans-18:1 isomer content and profile in edible refined beef tallow

Thetrans 18:1 acid content and profile for several samples of edible refined beef tallow were determined monthly over a period of one year. For this purpose, gas-liquid chromatography was combined with silver-ion thin-layer chromatography. The mean content oftrans-18:1 isomers was 4.9±0.9% (n=10) of total fatty acids with a minimum of 3.4% and a maximum of 6.2%. The distribution profile of individual isomers was also established. As in other ruminant fats (milk fat, meat fat), the main isomer is vaccenic (trans-11 18:1) acid. Other isomers, with their ethylenic bonds between positions 6 and 16, were found in lesser amounts. However, some slight but definite differences exist between beef tallow and cow milk fat. The relative proportion of vaccenic acid is higher in the former than in the latter. However, the distribution pattern oftrans-18:1 isomers in beef tallow closely resembles that in beef meat fat (lean part).     

Participation of thecis-12 ethylenic bond tocis-trans isomerization of thecis-9 andcis-15 ethylenic bonds in heated α-linolenic acid

To understand thecis-trans isomerization reaction of ethylenic bonds in heated octadecatrienoic acids (occurring during industrial deodorization of oils), we have prepared a mixture ofcis-9,cis-12,cis-15, andcis-9,cis-15 18:2 acids by partial hydrazine reduction ofcis-9,cis-12,cis-15 18:3 acid present in linseed oil. This mixture (as fatty acid methyl esters) was heated under vacuum at 270°C for 2.25 h. The two methylene-interrupted acids isomerize at a similar rate under such conditions, but the nonmethylene-interruptedcis-9,cis-15 18:2 acid remains unchanged. This means that the mechanism of isomerization does not involve a direct interaction between the two external ethylenic bonds as previously hypothesized. The centralcis-12 ethylenic bond is apparently necessary for the isomerization of the two externalcis-9 andcis-15 ethylenic bonds. However, this bond is itself rather protected against isomerization in the originalcis-9,cis-12,cis-15 18:3 acid which is mainly isomerized totrans-9,cis-12,trans-15,cis-9,cis-12,trans-15, andtrans-9,cis-12,cis-15 18:3 acids. Thecis-9,trans-12,cis-15 18:3 isomer is less than 10% of totaltrans isomers of α-linolenic acid. As a general rule, only one of the two double bonds in a methylene-interrupted diethylenic system can undergocis-trans isomerization when submitted to heat treatment, at least for temperatures equal to or less than 270°C.     

Triacylglycerols of winter butterfat containing configurational isomers of monoenoic fatty acyl residues. I. Disaturated monoenoic triacylglycerols

The triacylglycerols of winter butterfat were fractionated according to the type and degree of unsaturation into six fractions by silver ion high-performance liquid chromatography (Ag-HPLC). The acyl carbon number distribution of the triacylglycerols in each fraction was elucidated by reversed-phase HPLC and mass spectrometry (MS). The MS analysis of each fraction gave deprotonated triacylglycerol [M - H]⁻ ions which were produced by chemical ionization with ammonia. The daughter spectrum of each of the [M - H]⁻ ions provided information on its fatty acid constituents. Successful fractionation of triacylglycerols differing in the configuration of one fatty acyl residue by Ag-HPLC was important because geometrical isomers could not be distinguished by the MS system used. In addition to the fatty acid compositions, reversed-phase HPLC analysis demonstrated the purity of the collected fractions: molecules having acis-trans difference were separated nearly to the baseline. Triacylglycerols differing in the configuration of one fatty acyl residue were not equally distributed in relation to their acyl carbon numbers. This indicates that during the biosynthesis of triacylglycerols,cis- andtrans-fatty acids are processed differently. Although the fatty acid compositions of the corresponding molecular weight species of disaturatedtrans- and disaturatedcis-monoenoic triacylglycerols were similar, there may be differences in the amounts of different fatty acid combinations or in the distribution of fatty acids between the primary and secondary glycerol positions. In addition to the main components, it was possible to analyze minor triacylglycerols, such as molecules containing one odd-chain fatty acid, by the MS system used.