Influence of partially hydrogenated vegetable and marine oils on lipid metabolism in rat liver and heart

Partially hydrogenated marine oils containing 18∶1-, 20∶1- and 22∶1-isomers and partially hydrogenated peanut oil containing 18∶1-isomers were fed as 24–28 wt % of the diet with or without supplement of linoleic acid. Reference groups were fed peanut, soybean, or rapeseed oils with low or high erucic acid content. Dietary monoene isomers reduced the conversion of linoleic acid into arachidonic acid and the deposition of the latter in liver and heart phosphatidylcholine. This effect was more pronounced for the partially hydrogenated marine oils than for the partially hydrogenated peanut oil. The content oftrans fatty acids in liver phospholipids was similar in groups fed partially hydrogenated fats. The distribution of various phospholipids in heart and liver was unaffected by the dietary fat. The decrease in deposition of arachidonic acid in rats fed partially hydrogenated marine oils was shown in vitro to be a consequence of lower Δ6-desaturase activity rather than an increase in the peroxisomal β-oxidation of arachidonic acid. The lower amounts of arachidonic acid deposited may be a result of competition in the Δ6-desaturation not only from the C22-and C20-monoenoic fatty acids originally present in the partially hydrogenated marine oil, but also from C18- and C16-monoenes produced by peroxisomal β-oxidation of the long-chain fatty acids. Part of this work was presented at the ISF-AOCS Congress, New York City, 1980.     

The effect of dietary partially hydrogenated marine oils on desaturation of fatty acids in rat liver microsomes

The influence of dietary partially hydrogenated marine oils on distribution of phospholipid fatty acids in rat liver microsomes was studied with particular reference to the metabolism of linoleic acid. Five groups of weanling rats were fed diets containing 20% (w/w) peanut oil (PO), partially hydrogenated peanut oil (HPO), partially hydrogenated Norwegian capelin oil (HCO), partially hydrogenated herring oil (HHO), and rapeseed oil (RSO) for 10 weeks. The partially hydrogenated oils were supplemented with linoleic acid corresponding to 4.6 cal % in the diets. Accumulation of linoleic acid and reduced amount of total linoleic acid metabolites were observed in liver microsomal phospholipids from rats fed partially hydrogenated oils as compared to PO feeding. The most striking effects on the distribution of ω6-polyunsaturated fatty acids was obtained after feeding HHO, a marine oil with a moderate content oftrans fatty acids in comparison with HPO but rich in isomers of eicosenoic and docosenoic acids. Liver microsomal Δ⁶-as well as Δ⁶-desaturase activities as measured in vitro were reduced in rats kept on HHO as compared to PO dietary treatment. The results obtained suggest that the dietary influence of partially hydrogenated marine oils on the metabolism of linoleic acid might be better related to the intake of isomeric eicosenoic and docosenoic acids than to the total intake oftrans fatty acids.     

The effects of partially hydrogenated marine oils on the mitochondrial function and membrane phospholipid fatty acids in rat heart

The influence of dietary partially hydrogenated marine oils containing docosenoic acid on rat heart mitochondrial membrane phospholipid fatty acid composition was studied with particular reference to cardiolipin and oxidative phosphorylation. Five groups of male weanling rats were fed diets containing 20% (w/w) peanut oil (PO), partially hydrogenated peanut oil (HPO), partially hydrogenated Norwegian capelin oil (HCO), partially hydrogenated herring oil (HHO), and rapeseed oil (RSO) for 10 weeks. All the cardiac phospholipids investigated were influenced by the experimental diets. An increased amount of arachidonic acid observed in phosphatidylethanolamine (PE) after feeding partially hydrogenated oils suggests a changed regulation of the arachidonic acid metabolism in comparison with PO treatment. 22∶1 originating from the dietary oils was incorporated only to a small extent into phosphatidylcholine (PC) and PE. A selective incorporation of 18∶1 isomers into the 1- and 2-positions of PC and PE with respect to geometry and position of the double bond was observed. Large amounts of 18∶1trans were incorporated into the 1-position of PC and PE, irrespective of the amount of 18∶2 supplemented to the diets, replacing a considerable proportion of stearic acid in this position. After feeding HHO and RSO, the content of 22∶1 in mitochondrial cardiolipin of rat heart was found to be 3% (mainly cetoleic acid) and 10% (mainly erucic acid), respectively, indicating a high affinity forcis isomers of 22∶1, but also a considerable resistance against incorporation oftrans isomers was observed. The ability of rat cardiac mitochondria to oxidize palmitoylcarnitine and to synthesize ATP was depressed after feeding HHO and RSO. Dietarycis isomers of 22∶1 seem to have a specific ability to interfere with cardiac ATP synthesis and also to alter the fatty acid composition of cardiolipin of rat heart.     

Isomers of monoethylenic fatty acids in some partially hydrogenated marine oils

An analytical study of the monoethylenic isomers in commercial samples of partially hydrogenated herring, whale and seal oils is presented. The results show that with hydrogenated herring oil there is a slight decline in monoenetrans content from 37% in C₁₆ through to 32% in C₂₂. With both whale and seal oils, monoenetrans contents were constant at 54% and 59%, respectively, throughout all chain lengths. In general thecis andtrans positional isomers from hydrogenated whale and seal oils were more scattered than those from hydrogenated herring oil; however in each oil the majorcis isomers of each chainlength were indicative of originalcis fatty acid isomers in the raw oils.     

Effects of dietary proteins on linoleic acid desaturation and membrane fluidity in rat liver microsomes

The effect of dietary protein, casein (CAS) and soybean protein (SOY), on linoleic acid desaturation in liver microsomes was studied in rats. The activity of Δ6 desaturase in total and rough endoplasmic reticula (ER and RER) was significantly higher in the CAS group than in the SOY group. In ER and smooth endoplasmic reticulum, the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene, when incorporated into the membrane, was decreased in the SOY group and accompanied by a reduction in the cholesterol/phospholipid (CHOL/PL) ratio, consistent with an increase in membrane fluidity. In a separate study, the effect of varying dietary proteins, CAS, milk whey protein, egg albumin, SOY, potato protein and wheat gluten, on the relationship between the Δ6 desaturase activity and microsomal membrane fluidity was also examined. The results indicated that the dietary protein-dependent change in the liver microsomal CHOL/PL ratio affected membrane fluidity, and subsequently the activity of Δ6 desaturase in liver microsomes. However, since dietary protein influenced the Δ6 desaturase activity in RER without influencing membrane fluidity, it is possible that some regulation might have taken place at the level of enzyme synthesis.     

Dietary linoleic acid and the fatty acid profiles in rats fed partially hydrogenated marine oils

The influence of the linoleic acid levels of diets containing partially hydrogenated marine, oils (HMO) rich in isomeric 16∶1, 18∶1, 20∶1 and 22∶1 fatty acids on the fatty acid profiles of lipids from rat liver, heart and adipose tissue was examined. Five groups of rats were fed diets containing 20 wt% fat−16% HMO+4% vegetable oils. In these diets, the linoleic acid contents varied between 1.9% and 14.5% of the dietary fatty acids, whereas the contents oftrans fatty acids were 33% in all groups. A sixth group was fed a partially hydrogenated soybean oil (HSOY) diet containing 8% linoleic acid plus 32%trans fatty acids, mainly 18∶1, and a seventh group, 20% palm oil (PALM), with 10% linoleic acid and notrans fatty acids. As the level of linoleic acid in the HMO diets increased from 1.9% to 8.2%, the contents of (n−6) polyunsaturated fatty acids (PUFA) in the phospholipids increased correspondingly. At this dietary level of linoleic acid, a plateau in (n−6) PUFA was reached that was not affected by further increase in dietary 18∶2(n−6) up to 14.5%. Compared with the HSOY- or PALM-fed rats, the plateau value of 20∶4(n−6) were considerably lower and the contents of 18∶2(n−6) higher in liver phosphatidylcholines (PC) and heart PC. Heart phosphatidylethanolamines (PE) on the contrary, had elevated contents of 20∶4(n−6), but decreased 22∶5(n−6) compared with the PALM group. All groups fed HMO had similar contents oftrans fatty acids, mainly 16∶1 and 18∶1, in their phospholipids, irrespective of the dietary 18∶2 levels, and these contents were lower than in the HSOY group. High levels of linoleic acid consistently found in triglycerides of liver, heart and adipose tissue of rats fed HMO indicated that feeding HMO resulted in a reduction of the conversion of linoleic acid into long chain PUFA that could not be overcome by increasing the dietary level of linoleic acid.     

The deposition of polyunsaturated fatty acids in the rat fed partially hydrogenated vegetable oil

The composition of deposited polyenoic fatty acids in rats fed liquid or partially hydrogenated corn oil was determined by gas chromatography, which did not distinguish isomeric forms, and by lipoxidase which reacted with thecis, cis-methylene-interrupted acids. The two methods gave similar results for the liver fatty acids of rats fed either the unhydrogenated or partially hydrogenated oil. Of the fatty acids from the epididymal fat pads of rats fed the hydrogenated product, an appreciable quantity of linoleate isomers did not react with lipoxidase. The total amount of linoleic acid deposited was related to the total fatty acid pattern of the dietary oil. It appeared that thetrans-acids were mostly metab-olized and that the originalcis,cis-linoleic acid remaining in the partially hydrogenated product was preferentially incorporated into tissues. One of 10 papers to be published from the Symposium “Hydrogenation” presented at the AOCS Meeting, New Orleans, April 1970.     

Effect of dietary fatty acid composition on the biosynthesis of unsaturated fatty acids in rat liver microsomes

A study was made of the influence of semisynthetic diets of low and high unsaturation on the fatty acid composition and desaturation-chain elongation enzymatic activity of the liver microsomal fractions of male Sprague-Dawley rats of different ages. Groups of rats were fed 5 or 20% coconut oil (CO), or a 5 or 20% mixture of corn and menhaden oils (3∶7) (CME) from weaning to 100 wk of age. Growth rate and food consumption were measured during this period in which animals were sacrificed at 36, 57, 77 and 100 wk of age. Both the level and composition of the dietary fat supplements produced marked effects on the fatty acid composition of the liver microsomal lipids. In general, the fatty acid composition of the microsomal fractions reflected that of the dietary fat and was more unsaturated with the higher level of fat fed. The rate of conversion of linoleic to arachidonic acid in assays performed in vitro with liver microsomal preparations from animals of the different groups also showed marked differences. The 6-desaturase-chain elongation activity was higher in the 5% than 20% group and corresponded to the essential fatty acid (EFA) status of the animals in these groups as represented by the triene-tetraene ratio of the microsomal lipid. The relationship of the 6-desaturase activity to fatty acid composition of the microsomal lipid indicated that if varied directly with the level of 20∶3ω9, 18∶1 and 16∶1 and was inhibited by arachidonic acid. The activity of the 6-desaturase enzyme system was lowest in the liver microsomal fraction obtained from the animals fed the CME diets and appeared to be suppressed by the high levels of 20∶5 and 22∶6 that accumulated in the microsomal lipid. Accordingly, the levels of arachidonic acid were lower in the microsomal lipid of these groups than those of the corresponding CO groups in spite of a greater abundance of linoleic acid in the diet. The data suggest that the activity of the 6-desaturase-chain elongation system is regulated by the fatty acid composition of the microsomal lipid as influenced by the composition of the dietary fat.     

Myocardial alterations resulting from feeding partially hydrogenated marine oils and peanut oil to rats

Myocardial alterations were observed in 5 groups of rats fed diets containing 20% fat for 16 weeks. The incidence was comparable to that from other studies and uniform at 6/20 in hearts from rats fed: partially hydrogenated herring oil to give dietary levels of either 16.7% or 4.6% 22∶1; partially hydrogenated redfish/flatfish oil to give 4.5% 22∶1 in the dietary fat; and peanut oil (of unknown origin) containing 0.1% 22∶1. The incidence was 9/20 in the hearts of rats fed an unrefined and unprocessed redfish oil at a dietary level of 16.0% 22∶1 in fatty acids.     

Application of methoxy-bromomercuri-adduct fractionation to the analysis of fatty acids of partially hydrogenated marine oils

The fatty acids of a refined and of a partially hydrogenated menhaden oil, iodine value (IV) 84.5, were separated into different classes (e.g., monoene, diene, including pentaene and hexaene) by thin layer chromatography (TLC) of their methoxy-bromomercuri-adducts (MBM). In the solvent system hexane: dioxane, the separation of fatty acids occurred according to the degree of unsaturation. No influence was exerted by either the geometry or the position of the ethylenic bonds. The effect of the various chain lengths (C₁₄−C₂₂) was to broaden the bands, but no overlap occurred among the chain lengths. A wide range of C₂₀ unsaturated fatty acids were prepared by the hydrazine reduction of 20∶5-Δ5,8,11,14,17. These were separated into groups as MBM adducts and identified by comparison of their experimental and calculated equivalent chain lengths (ECL) in gas liquid chromatography (GLC) on SILAR-5CP and SILAR-7CP columns. This confirmed that GLC did not totally separate all groups of isomers of different degrees of unsaturation. The quantitative analysis of both refined and partially hydrogenated (IV-84.5) menhaden oils by GLC was effected by the recovery of the fatty acid methyl esters from the MBM adduct TLC bands with the addition of methyl heptadecanoate (17∶0) as an internal standard, followed by analysis of the different fractions on open-tubular columns coated with SILAR-5CP. For methylene- and nonmethylene-interrupted unsaturated acids, 100% recovery from the MBM adducts was achieved, but in the case of the conjugated dienes the maximal recovery was 70%.     

Determination of partially hydrogenated terphenyls-based thermal heating fluid in vegetable oils by HPLC with fluorescence detection

An HPLC method for the determination of partially hydrogenated terphenyls-based thermal heating fluid, Therminol 66^™, in various vegetable oils is described. Direct analysis of palm olein showed that the 3- and 4-cyclohexylbiphenyl peaks of the Therminol 66^™ used in quantitative analysis co-eluted with other fluorescent peaks present naturally in the oil. However, those interfering peaks were readily removed after saponification of palm olein. The concentrations of the 3- and 4-cyclohexylbiphenyls of Therminol 66^™ were monitored by fluorescence detection at 257 (excitation) and 320 nm (emission). The calibration graph obtained by using the peak areas of the 3- and 4-cyclohexylbiphenyls against the concentrations of Therminol 66^™ was linear, with a correlation coefficient of 0.994. The limit of quantitation, using spiked palm olein, was as low as 0.2 μg/g. The coefficients of variation obtained from the intra- and interday studies obtained by using three spiked concentrations (0.2, 0.5, and 1.0 μg/g) were 1.76–6.43 and 3.77–10.4%, respectively. The mean recovery value obtained from sunflower, soybean, and canola oils was more than 88.7%.     

Determination of ultratrace metals in hydrogenated vegetable oils and fats

Ultratrace levels of nickel, chromium, copper and iron occurring in hydrogenated vegetable oil products were estimated by dispersion of the samples in 4 methyl-2-pentanone and atomic absorption analysis by the graphite furnace technique. The principal goals in establishing the analytical methods were improved sensitivity to metals at low levels and applicability to limited amounts of products. Using reproducibility and linearity of response as criteria, optimum oil concentration in solvent and instrument parameters were established. For a series of commercial products, the method of standard addi-tions was adopted to correct for matrix differences between the products and salad oil-based standards. The range for the metals was determined in five cooking oils: Ni, 29–207 ppb; Cr, 1–5 ppb; Cu, 13–37 ppb; and Fe, 138–301 ppb; in recovered oils from five margarines: Ni, 34–70 ppb; Cr, 2–12 ppb; Cu, 26–58 ppb; and Fe, 239–540 ppb; and in five solid shortenings: Ni, 592–2772 ppb; Cr, 8–35 ppb; Cu, 26–108 ppb.     

Pan-heating of low-linolenic acid and partially hydrogenated soybean oils

Genetically modified low-linolenic acid soybean oil (LL-SBO) was compared to partially hydrogenated soybean oil (PH-SBO). Samples were heated on a Teflon pan at ∼180°C until a selected end point of ≥20% polymer content was reached. High-performance size-exclusion chromatography analysis indicated the PH-SBO contained >20% polymer after 20 min of heating, whereas the LL-SBO sample contained >20% polymer after 10 min. Supercritical fluid chromatography analysis indicated degradation rates of 0.161±0.011 min⁻¹ for LL-SBO and 0.086±0.004 min⁻¹ for PH-SBO. The volatile compounds were identified and quantitated with static head-space-GC-MS. 1-Heptene (239.9 ppm) and hexanal (1486.1 ppm) were present at the greates concentration among the volatile compounds in LL-SBO. The volatile compounds present in the greatest concentrations in heated PH-SBO were hexanal (376.9 ppm) and pentane (82.1 ppm). After 10 min of heating, the LL-SBO oil FFA value (2.66%), p-anisidine value (386.5 abs/g oil), Food Oil Sensor reading (18.75), and color intensity (Y=4.0, R=1.0) were significantly greater than those of PH-SBO after 14 min of heating (4.28%, 298.5 abs/g oil, 16.08, Y=1.0, R=0.1, respectively). There was a significant difference in the degradation rates between LL-SBO and PH-SBO (P<0.05). The PH-SBO was more stable than the LL-SBO.     

Fate of dietary sterols in hydrogenated oils and fats

Samples of table margarines, so-called polyunsaturated table margarines, hydrogenated vegetable oils, and so-called polyunsaturated hydrogenated vegetable oils were shown by infrared spectroscopy to contain hydrogenated components. Examination of the sterols from these oils by argentation thin layer chromatography and gas liquid chromatography did not reveal campestanol, stigmastanol, or Δ²²-stigmastenol, the expected hydrogenation products of the natural sterols. The sterol compositions of the above samples, animal fats, and blends of hydrogenated vegetable oils and animal fats were determined. The compound 24-methyl cholest-7-en-3β-ol was identified tentatively in sunflower and safflower oils.     

Turnover of fatty acyl-CoA oxidase in the liver of rats fed on a partially hydrogenated marine oil

The change in turnover of fatty acyl-CoA oxidase (FAO), the rate-limiting enzyme of the peroxisomal β-oxidation system, was investigated in rats fed a 30% (w/w) partially hydrogenated marine oil (PHMO) diet. The FAO activity increased five-fold after two weeks of PHMO feeding, and decreased after withdrawal of the diet. Based onin vivo experiments using L-[4,5-³H]leucine and an immunoprecipitation technique, the increase in the activity of FAO could be accounted for by a 1.6-fold higher rate of FAO synthesis and a 3.4-fold slower rate of FAO degradation as compared to controls. In the same PHMO-fed rats, the rates of synthesis and degradation of carnitine palmitoyltransferase were 1.8-fold higher and 2.0-fold slower, respectively, as compared to controls. The results indicate that the observed increase in the activity of the enzymes of peroxisomal β-oxidation is mainly due to a reduced rate of FAO degradation in the liver of rats fed the PHMO diet.     

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.     

Metabolism of erucic acid in perfused rat liver: Increased chain shortening after feeding partially hydrogenated marine oil and rapeseed oil

The metabolism of [14-¹⁴C] erucic acid was studied in perfused livers from rats fed on diets containing partially hydrogenated marine oil or rapeseed oil for three days or three weeks. Control rats were given groundnut oil. Chain-shortening of erucic acid, mainly to 18∶1, was found in all dietary groups. In the marine oil and rapeseed oil groups, the percentage of chain-shortened fatty acids in very low density lipoproteins-triacylglycerols (VLDL-TG) exported from the liver increased after prolonged feeding. A similar increase was found in liver TG only with partially hydrogenated marine oil. This oil, rich intrans fatty acids, thus seemed to be more effective in promoting chain-shortening. The fatty acid composition of the secreted and stored TG differed both with respect to total fatty acids and radioactively labeled fatty acids, indicating that at least 2 different pools of TG exist in the liver. The lack of lipidosis in livers from rats fed dietary oils rich in 22∶1 fatty acids is discussed in relation to these findings. In conclusion, a discussion is presented expressing the view that the reversal of the acute lipidosis in the hearts of rats fed rapeseed oil or partially hydrogenated marine oils is, to a large extent, derived from the increased chain-shortening capacity of erucic acid in liver.