Density‐Dependent Formation of the Pure trans‐Isomer of the Unsaturated Alcohol by Selective Hydrogenation of Citral in Supercritical Carbon Dioxide

Selective hydrogenation of citral to unsaturated alcohol [geraniol (trans) + nerol (cis)] was carried out in supercritical carbon dioxide (scCO₂) using an MCM‐41 supported plantinum catalyst (∼1 wt% Pt). A remarkable rate of isomerization of the unsaturated alcohol [nerol (cis) to geraniol (trans)] during the hydrogenation of citral was achieved simply by tuning the density of CO₂. Optimum reaction conditions were developed to obtain only geraniol (trans) with a selectivity of 98.8% and citral conversion of 99.8%. A significant change in the cis:trans ratio of the product (1:82.3) from the substrate (1:1.3) was observed depending on the various reaction parameters like carbon dioxide and hydrogen pressure, reactant concentration, reaction time and, particularly, the total selectivity for unsaturated alcohol [geraniol (trans) +nerol (cis)]. It has been observed that the presence of hydrogen is necessary for isomerization. Our results were explained in terms of a density‐dependent, two‐step model. The kinetic behaviour shows that the rate of isomerization was higher in scCO₂ compared to other organic solvents and the pure form of geraniol (trans) was obtained exclusively. A probable reaction pathway was proposed in order to explain the isomerization during hydrogenation of citral in scCO₂ medium.     

Directed sequential synthesis of conjugated linoleic acid isomers from Δ7, 9 to Δ12, 14

Position and configuration isomers of conjugated linoleic acid (CLA), from 7, 9‐ through 12, 14‐C18:2, were synthesized by directed sequential isomerizations of a mixture of rumenic (cis‐9, trans‐11 C18:2) and trans‐10, cis‐12 C18:2 acids. Indeed, the synthesized conjugated fatty acids cover the range of unsaturated systems as found in milk fat CLA. The two‐step sequence consisted in initial sigmatropic rearrangement of cis/trans CLA isomers at 200 °C for 13 h under inert atmosphere (Helium, He), followed by selenium‐catalyzed geometrical isomerization of double bonds at 120 °C for 20 h under He. Product analysis was achieved by gas‐liquid chromatography using a 120 m polar capillary column coated with 70% cyanoalkylpolysiloxane equivalent polymer. Migration of conjugated systems was geometrically controlled as follows: the cis‐C_n, trans‐C_n+2 double bond system was rearranged through a pericyclic [1, 5] sigmatropic mechanism into a trans‐C_n‐1, cis‐C_n+1 unsaturated system, while the trans‐C_n, cis‐C_n+2 double bond system was rearranged through a similar pericyclic mechanism into a cis‐C_n+1, trans‐C_n+3 unsaturated system. Selenium‐catalyzed geometrical isomerization under mild conditions then allowed cis/trans double bond configuration transitions, resulting in the formation of all cis, all trans, cis‐trans and trans‐cis isomers. A sequential combination of the two reactions resulted in a facile controlled synthesis of CLA isomers, useful for the chromatographic identification of milk fat CLA, as well as for the preparation of CLA standard mixture.     

cis/trans isomerization of unsaturated fatty acids as possible control mechanism of membrane fluidity inPseudomonas putida P8

Exponentially growing cells ofPseudomonas putida had an increased ratio of saturated to unsaturated fatty acids in response to increased growth temperatures. Resting cells in which fatty acid biosynthesis was stopped reacted to a thermal increase by convertingcis-monounsaturated fatty acids totrans isomers.cis/trans isomerization of up to 60% of the unsaturated fatty acids was also activated by alcohols of different chain length. Their effective concentrations apparently depended on the lipophilic character of the alcohols. Also, a salt shock caused by the addition of NaCl resulted in the production oftrans fatty acids. However, cells that were adapted to growth media of high osmolarity synthesized cyclopropane fatty acids instead oftrans fatty acids. Activity ofcis/trans-isomerase was dependent on the growth phase and was significantly higher during logarithmic growth than during the stationary phase. The results of this study agree with the hypothesis that the isomerization ofcis intotrans unsaturated fatty acids is an emergency action of cells ofP. putida to adapt membrane fluidity to drastic changes of environmental conditions.     

Isomerization of unsaturated fatty esters by iron pentacarbonyl. Preparation of iron tricarbonyl complexes of polyunsaturated fats

Iron pentacarbonyl is a powerful isomerization agent of unsaturated fatty esters. Highly conjugated fats are obtained when polyunsaturated fatty esters are treated with an excess Fe(CO)₅ to form complexes followed by decomposition of the complexes with FeCl₃. Iron tricarbonyl complexes were prepared in 80 to 95% yields from methyl linoleate, linolenate and polyunsaturated fatty esters of soybean, linseed and safflower oils by heating at 180–185C with 2 moles Fe(CO)₅ per mole ester under nitrogen pressure. Decomposition of these complexes with FeCl₃ resulted in 90 to 97% conjugation of the polyunsaturated fatty esters mainly in the alltrans configuration. Isolatedtrans unsaturation reached levels of 18 to 30%. Methyl oleate yielded 74%trans unsaturation but no complex of iron carbonyl was obtained. Presented in part at AOCS meeting in Houston, 1965. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Allylic halogenation and oxidation of long chain α,β-unsaturated ester

Methyltrans-2-hexadecenoate (1b) on allylic bromination with N-bromosuccinimide (NBS) (0.5 mole) yielded methyl 4-bromo-trans-2-hexadecenoate (2b) in 50% yield. Reaction with 2.0 moles of NBS afforded the allylic bromide (2b, δ 80%) as well as the dibromide (3b). Alkaline hydrolysis of the bromo ester (2b) yielded 4-hydroxy-trans-2-hexadecenoic acid (4a, δ 70%) and an unexpected product, 4-oxo-hexadecanoic acid (5a, δ 30%). A mechanism involving rearrangement is proposed to account for this unusual product. The effect of the ester carbonyl adjacent to the double bond in2b was found to suppress dehydrobromination (elimination) reaction and favors only substitution (S_N2), unlike the allylically brominated derivatives of internal olefinic compounds. The CrO₃-pyridine oxidation of4b yielded the corresponding unsaturated ketone (7b). The structures of individual reaction products were established by elemental analyses as well as by spectral studies.     

Components of the hardening flavor present in hardened linseed oil and soybean oil

The characteristic hardening flavor which develops in hardened linseed and soybean oils during storage has been coned from hardened linseed oil by stripping with nitrogen. After separating the volatile substances by adsorption chromatography on silica, the fraction containing the hardening flavor has been converted into 2,4-dinitrophenylhydrazones (DNPHs) and separated by means of partition chromatography. On regeneration of the fractions of DNPSs obtained, the characteristic hardening flavor was observed in one specific band. Both by hydrogenation and by oxidation of the free carbonyl the carrier of the flavor was found to be an unsaturated aldehyde; however, not of the α-β unsaturated type. Further separation of the regenerated carbonyls by means of gas-liquid chromatography (GLC) points to a C₉-aldehyde. After synthesis of the 4-,5- and 6-cis- andtrans-nonenals, comparison made it probable that the carrier of the hardening flavor is a mixture of 6-cis and 6-trans-nonenal, the latter of which has the greatest share in the hardening flavor. In order to confirm the location of the double bond in the carrier of the hardening flavor a recent isolation technique was applied. The volatile substances from hardened linseed oil were first separated via GLC. After conversion of the carbonyls in question into their DNPHs, the latter have been separated by means of thin-layer chromatography (TLC). By means of IR-analysis and oxidation with osmium tetroxide of the pure derivative, the principal carrier of the hardening flavor has been identified as 6-trans-nonenal.     

Trans unsaturated fatty acids in bacteria

The occurrence oftrans unsaturated fatty acids as by-products of fatty acid transformations carried out by the obligate anaerobic ruminal microflora has been well known for a long time. In recent years, fatty acids withtrans configurations also have been detected in the membrane lipids of various aerobic bacteria. Besides several psychrophilic organisms, bacteria-degrading pollutants, such asPseudomonas putida, are able to synthesize these compoundsde novo. In contrast to thetrans fatty acids formed by rumen bacteria, the membrane constituents of aerobic bacteria are synthesized by a direct isomerization of the complementarycis configuration of the double bond without a shift of the position. This system of isomerization is located in the cytoplasmic membrane. The conversion ofcis unsaturated fatty acids totrans changes the membrane fluidity in response to environmental stimuli, particularly where growth is inhibited due to the presence of high concentrations of toxic substances. Under these conditions, lipid synthesis also stops so that the cells are not able to modify their membrane fluidity by any other mechanism.     

Cis-Unsaturated fatty acid products by hydrogenation with chromium hexacarbonyl

The use of Cr(CO)₆ was investigated to convert polyunsaturated fats intocis unsaturated products. With methyl sorbate, the same order of selectivity for the formation ofcis-3-hexenoate was demonstrated for Cr(CO)₆ as for the arene-Cr(CO)₃ complexes. With conjugated fatty esters, the stereoselectivity of Cr(CO)₆ toward thetrans, trans diene system was particularly high in acetone. However, this solvent was not suitable at elevated temperatures required to hydrogenatecis, trans- andcis, cis-conjugated dienes (175 C) and nonconjugated soybean oil (200 C). Reaction parameters were analyzed statistically to optimize hydrogenation of methyl sorbate and soybean oil. To achieve acceptable oxidative stability, it is necessary to reduce the linolenate constituent of soybean oil below 1–3%. When this is done commercially with conventional heterogenous catalysts, the hydrogenated products contain more than 15%trans unsaturation. By hydrogenating soybean oil with Cr(CO)₆ (200 C, 500 psi H₂, 1% catalyst in hexane solution), the product contains less than 3% each of linolenate andtrans unsaturation. Recycling of Cr(CO)₆ catalyst by sublimation was carried through three hydrogenations of soybean oil, but, about 10% of the chromium was lost in each cycle by decomposition. The hydrogenation mechanism of Cr(CO)₆ is compared with that of arene-Cr(CO)₃ complexes. Presented in part at Seventh Conference on Catalysis in Organic Syntheses, Chicago, Illinois, June 5–7, 1978.     

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.     

13C N.m.r. Spectra of ring-opened polymers of 1-methylbicyclo[2.2.1]hept-2-ene and their hydrogenated products

The ¹³C n.m.r. spectra of the title polymers have been completely assigned for a comprehensive range of cis contents (σ_c = 0–1) and head-tail bias (HT/(HH + TT) = 1 −∞). In the unsaturated polymers (II), cis HH dyads are never found, so that all-cis polymers are also all-Ht, but cis TT dyads are formed in cis-trans polymers, equal to the difference between trans HH and trans TT. The olefinic region consists of seven triplets; two each from cis HT and trans HT, and one each from cis TT, trans TT and trans HH. In the case of trans each triplet consists of a main centre peak due to ttt and ctc sequences and two wing peaks due to ttc and ctt sequences, as defined in the text. The upfield peaks also show cc/ct or tt/tc splittings as well as HH/HT or TT/TH splittings in most cases. In the saturated polymers C⁶ is sensitive to m/r ring dyad sequences. A corrected assignment is given for the hydrogenated polymer of norbornene.     

A rapid method for the determination oftrans unsaturation in fats and derivatives

Thetrans unsaturation present in samples of unsaturated fats and derivatives may be determined from measurement of the infrared absorbance at two wavelengths, one due totrans configuration of a double bond and the other due to the ester or acid group. The ratio of the two absorbance values has a linear relationship with thetrans unsaturation content. Thus, a drop of sample is diluted with CS₂ and the infrared absorbance measured at 10.36 μ and at 8.5–8.6 μ if esters and 10.7 μ if acids. To calculate the per cent isolatedtrans double bonds in the sample, the ratio of the two absorbance values is substituted into the linear equation which was developed from known samples. The method is rapid since the sample need not be weighed nor made up to a known volume and thus may be applied to samples such as those collected from a gas chromatograph or after thin layer chromatography. Presented at the AOCS Meeting, New York, October 1968.     

Direct determination oftrans unsaturation in triglycerides by infrared spectrophotometry

A differential infrared spectrophotometric method is described for the determination oftrans unsaturation in fats. The method utilizes absorption at 965 cm⁻¹, due to the C−H out-of-plane deformation vibrations oftrans unsaturated compounds. The method is rapid, accurate, and directly applicable to the determination oftrans unsaturation in triglycerides. It is applicable to samples which contain low concentrations oftrans acids (down to 2%) and also to samples with fatty acids of mixed chain length.     

A comparative study on the selective hydrogenation of α,β unsaturated aldehyde and ketone to unsaturated alcohols on Au supported catalysts

The hydrogenation of trans,4-phenyl,3-buten,2-one (benzalacetone) and trans,3-phenyl, propenal (cinnamaldehyde) was carried out on Au supported on iron oxides catalysts. Commercial goethite (FeOOH), maghemite (γFe₂O₃) and hematite (αFe₂O₃) were used as supports. The catalytic activity of Au/Fe₂O₃ reference catalyst, supplied by the World Gold Council, was also investigated. Gold catalysts and the parent supports were characterized by BET, X-ray diffraction (XRD), temperature programmed reduction (TPR), temperature programmed desorption of ammonia (NH₃-TPD) and high resolution transmission electron microscopy (HRTEM).Among the catalysts investigated Au supported on FeOOH shows the highest activity and selectivity to UA in the hydrogenation of unsaturated carbonyl compounds whereas Au supported on αFe₂O₃ are the less active and selective catalysts.The catalytic activity and selectivity to unsaturated alcohols (UA) in the hydrogenation of benzalacetone and cinnamaldehyde are less influenced by the morphology of gold particles and are mainly influenced by the nature of the support.A correlation between the reducibility of the catalysts and the activity and selectivity to UA has been found. Increasing the reducibility of the catalysts both the activity and selectivity to UA increase. These results let us to argue that active and selective sites are formed by negative gold particles formed through the electron transfer from the reduced support to the metal.     

Trans fatty acid isomers in Canadian human milk

The fatty acid composition, totaltrans content (i.e., sum of all the fatty acids which may have one or moretrans double bonds) and geometric and positional isomer distribution of unsaturated fatty acids of 198 human milk samples collected in 1992 from nine provinces of Canada were determined using a combination of capillary gas-liquid chromatography and silver nitrate thin-layer chromatography. The mean totaltrans fatty acid content was 7.19±3.03% of the total milk fatty acids and ranged from 0.10 to 17.15%. Twenty-five of the 198 samples contained more than 10% totaltrans fatty acids, and thirteen samples contained less than 4%. Totaltrans isomers of linoleic acid were 0.89% of the total milk fatty acids with 18∶2Δ9c, 13t being the most prevalent isomer, followed by 18∶2Δ9c, 12t and 18∶2Δ9t, 12c. Using the totaltrans values in human milk determined in the present study, the intake of totaltrans fatty acids from various dietary sources by Canadian lactating women was estimated to be 10.6±3.7 g/person/d, and in some individuals, the intake could be as high as 20.3 g/d. The 18∶1trans isomer distribution differed from that of cow's milk fat but was remarkably similar to that in partially hydrogenated soybean and canola oils, suggesting that partially hydrogenated vegetable oils are the major source of thesetrans fatty acids.     

<Emphasis Type="Italic">Trans</Emphasis> fatty acids and atopic eczema/dermatitis syndrome: The relationship with a free radical <Emphasis Type="Italic">cis-trans</Emphasis> isomerization of membrane lipids

The formation of trans FA residues in membrane phospholipids may be due to a free radical-catalyzed isomerization process occurring to the cis unsaturated FA moieties. Radical stress is well documented in inflammatory processes of atopic diseases, but no data are yet available about a possible association with trans FA detected in these patients. We investigated the presence of trans lipid isomers in the erythrocyte and T-lymphocyte membranes of 26 children affected by atopic eczema/dermatitis syndrome (AEDS). Trans lipid isomers were found in both cell membranes, up to a total content of 2.7 and 4.9% of the FA composition, respectively. By using the geometrical trans lipid library derived from in vitro models of thiyl radical-catalyzed isomerization, oleic and arachidonic acid isomers were detected. The statistical significance was evaluated by comparison with an age-matched control group. These results suggest the role of an endogenous free radical isomerization path occurring to membrane unsaturated lipids, complementary to the dietary contribution, which can be involved in the lipid impairment in AEDS. This study contributes to lipidomic research regarding the double bond structure and the influence of a geometrical change of membrane lipids in physiology and diseases.     

Measurement oftrans and other isomeric unsaturated fatty acids in butterand margarine

A procedure is described for gas liquid chromatographic determination of cis andtrans isomers of unsaturated fatty acids after fractionation of the saturated, monenoic, dienoic, and polyenoic fatty acid methyl esters by argentation thin layer chromatography. To test its reliability, the procedure was used for quantitative measurement of transisomers of unsaturated fatty acids in a known mixture of simple triglycerides containing saturated fatty acids from 4:0 to 24:0 andcis andtrans isomers of 14:1. 16:1, 18:1, and 18:2. Results of the analyses of five margarine and five butter samples are presented, together with results of infrared spectrophotometric analyses fortrans fatty acid concentrations, ultraviolet spectrophotometric analyses for conjugated fatty acid concentrations, and enzymatic analyses forcis-cis-methylene interrupted fatty acid concentrations. The combined argentation thin layer and gas Chromatographic procedure is suitable for determination of the principal fatty acids in complex food lipids such as milk fat.     

Ring location in cyclopropane fatty acid esters by boron trifluoride-catalyzed methoxylation followed by mass spectroscopy

Methyl esters of methylcis- andtrans-9,10-methyleneoctadecanoic acids react with 50% boron trifluoride-methanol to produce unsaturated and methoxy-esters; both products are shown by gas chromatography to be a mixture of several isomers. Mass spectra of the methoxylated esters are characterized by intense peaks due to cleavage adjacent to methoxy-functions which allow the position of the ring in the original cyclopropane ester to be easily assigned. Methyl oleate is also partially attacked by 50% BF₃−MeOH to produce a mixture of methyl 9- and 10-methoxyoctadecanoates. 14% BF₃−MeOH does not react with cyclopropane and olefinic esters under the reaction conditions employed.     

Thiyl radical-catalyzed isomerization of oils: An entry to the trans lipid library

The unsaturated fatty acyl moieties of TAG present in natural oils of borage, olive, and rice were converted to their corresponding geometrical trans isomers by thiyl radical-catalyzed isomerization. Thiyl radicals were generated from 2-mercaptoethanol under photolytic or thermal conditions. A relevant feature of this method is the absence of double-bond shifts, so that no positional trans isomers or conjugated polyenes are formed. Oils obtained after the isomerization were winterized to further increase their trans fatty acid content. Methanolysis and hydrolysis of the trans oil mixtures using an enzymatic method (lipase B from Candida antarctica) gave good conversions to the corresponding trans FAME and fatty acids, respectively. These results are relevant for the studies of lipid isomerism and trans fatty acid recognition, which is a growing concern in biochemistry and nutrition, and open new perspectives for the synthesis of glycerides and studies of their structure-activity relationships.     

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.     

The use of SP2340 glass capillary columns for the estimation of thetrans fatty acid content of foods

Glass capillary gas chromatography (GCGC) on 100-m and 60-m SP2340 columns was used for quantitation of thetrans unsaturated fatty acids in shortenings and fast foods. The separation of thecis andtrans octadecenoates on GCGC was evaluated by preparatory argentation thin layer chromatography. In addition, thetrans content of shortening samples obtained by GCGC was compared totrans content determined by infrared analysis. This research conducted by the Science and Education Administration, USDA, on the commercial foods as reported in this paper was limited to analysis of their lipid compositions. The data are reported solely as factual information and are limited to the samples analyzed. No warranty or guarantee is made or implied that other samples of these products will have the same or similar composition. It is the policy of the USDA not to endorse those commercial products used in research over those that were not included in the research.