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Utilization of amylose-lipid complexes as molecular nanocapsules for conjugated linoleic Acid.

Amylose-conjugated linoleic acid (CLA) complexes were produced by water/dimethyl sulfoxide (DMSO) and KOH/HCl complexation methods. The formation of amylose V form was confirmed by X-ray diffraction (XRD), and complexes formed at 30, 60, and 90 degrees C exhibit melting temperatures exceeding 88 degrees C. Atomic force microscopy (AFM) images showed distinct difference in complex organization, with complexes formed in water/DMSO showing spherical shape with typical diameter of 150 nm. Complexes formed by KOH/HCl showed elongated structure with typical width of 43-160 nm. Water/DMSO complexes exhibit superior protection to CLA against oxidation. All complexes showed high retention of CLA in simulated stomach conditions, and the digestion of complexes by amylases results in high hydrolysis and CLA release by pancreatin and alpha-amylase. Only moderate release was detected following hydrolysis by amyloglucosidase and beta-amylase. It is therefore suggested that amylose-CLA complexes can serve as molecular nanocapsules for protection and delivery of CLA.

Amylases↗

Conjugated linoleic acid: a review.

Conjugated linoleic acid (CLA) refers to a group of positional and geometric isomers of the omega-6 essential fatty acid linoleic acid (cis-9, cis-12, octadecadienoic acid). In humans evidence is currently ambiguous as to whether CLA supplementation has a significant effect on body composition. Despite favorable changes in lipid levels in animal models, a beneficial effect in humans has not yet been established. While some of the changes reported are consistent with an improved lipid profile, declines in HDL and increases in lipoprotein (a) have also been observed in some subjects. Available evidence suggests CLA supplementation has no impact on immune system performance in healthy subjects.

Animals↗

Synthesis, isolation, and GC analysis of all the 6,8- to 13,15-cis/trans conjugated linoleic acid isomers.

Octadecadienoic acids with conjugated double bonds are often referred to as conjugated linoleic acid, or CLA. CLA is of considerable interest because of potentially beneficial effects reported from animal studies. Analysis of CLA is usually carried out by GC elution of FAME. If the presence of low-level isomers is of interest, a complementary technique such as silverion HPLC is also used. These analyses have been hindered by a lack of well-characterized commercially available reference materials. Described here are the synthesis and isolation of selected 6,8- through 13,15-positional CLA isomers, followed by isomerization of these CLA isomers with iodine to produce all the possible cis,cis, cis,trans, trans,cis, and trans,trans combinations. Also present are the GC retention times of the CLA FAME relative to gamma-linolenic acid (6c,9c,12c-octadecatrienoic acid) FAME using a 100-m CP Sil-88 capillary column (Varian Inc., Lake Forest, CA). These data include all the CLA isomers that have been identified thus far in foods and dietary supplements and should greatly aid in the future analysis of CLA in these products.

Chromatography, High Pressure Liquid↗

[Linoleic conjugated acid: current interest in human nutrition].

Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid. The major dietary sources of CLA are meat from rumiants and dairy products. There are also foods enriched in CLA (by modification animal feed or addition of commercial CLA from cartamo oil). However, the major isomer of CLA in natural food is cis-9,trans-11 and commercial CLA contains approximately equal amounts of cis-9,trans-11 and trans-10,cis-12 isomers. Although CLA is minor component of the diet, at present CLA has provoked intense scientific research accompanied by an advertising boom. This is due to the potential health effects demonstrated in animal and cell culture experiments including effects on body composition, specially a reduction in body fat mass and anticarcinogenic, antidiabetogenic and immune modulating effects. The results obtained in human volunteers are limited and even sometimes contradictory. This review focuses on recently studies in humans. To draw conclusions on CLA's potential interest as a functional ingredient, additional studies are needed to clearly define optimal level of CLA intake, short and long term effects and side effects of each individual CLA isomer in order to determine its safety and efficacy.

Body Composition↗

Gas chromatography-high resolution selected-ion mass spectrometric identification of trace 21:0 and 20:2 fatty acids eluting with conjugated linoleic acid isomers.

High-resolution selected-ion recording (SIR) of the exact molecular ion mass was used to confirm unambiguously the presence of conjugated linoleic acid (CLA) derivatives in biological matrices and standard mixtures and to differentiate non-CLA derivatives from CLA derivatives in the CLA region of the gas chromatogram. The success of this method was based on the selectivity of the SIR technique and its sensitivity, which was comparable to that of flame-ionization detection. A minor fatty acid methyl ester (FAME) was identified as methyl heneicosanoate (21:0), and six isomers of 20:2 FAME were found to elute in the CLA region. Isomerization of a standard CLA mixture resulted in a non-CLA flame-ionization response eluting in the CLA region of the gas chromatogram. It is therefore recommended that the identification of minor CLA isomers in natural products or biological matrices should include their direct confirmation by mass spectrometry.

Esters↗

Modulation of arachidonic acid distribution by conjugated linoleic acid isomers and linoleic acid in MCF-7 and SW480 cancer cells.

The relationship between growth and alterations in arachidonic acid (AA) metabolism in human breast (MCF-7) and colon (SW480) cancer cells was studied. Four different fatty acid preparations were evaluated: a mixture of conjugated linoleic acid (CLA) isomers (c9,t11, t10,c12, c11,t13, and minor amounts of other isomers), the pure c9,t11-CLA isomer, the pure t10,c12-CLA isomer, and linoleic acid (LA) (all at a lipid concentration of 16 microg/mL). 14C-AA uptake into the monoglyceride fraction of MCF-7 cells was significantly increased following 24 h incubation with the CLA mixture (P < 0.05) and c9,t11-CLA (P < 0.02). In contrast to the MCF-7 cells, 14C-AA uptake into the triglyceride fraction of the SW480 cells was increased while uptake into the phospholipids was reduced following treatment with the CLA mixture (P < 0.02) and c9,t11-CLA (P < 0.05). Distribution of 14C-AA among phospholipid classes was altered by CLA treatments in both cell lines. The c9,t11-CLA isomer decreased (P < 0.05) uptake of 14C-AA into phosphatidylcholine while increasing (P < 0.05) uptake into phosphatidylethanolamine in both cell lines. Both the CLA mixture and the t10,c12-CLA isomer increased (P < 0.01) uptake of 14C-AA into phosphatidylserine in the SW480 cells but had no effect on this phospholipid in the MCF-7 cells. Release of 14C-AA derivatives was not altered by CLA treatments but was increased (P < 0.05) by LA in the SW480 cell line. The CLA mixture of isomers and c9,t11-CLA isomer inhibited 14C-AA conversion to 14C-prostaglandin E2 (PGE2) by 20-30% (P < 0.05) while increasing 14C-PGF2alpha by 17-44% relative to controls in both cell lines. LA significantly (P < 0.05) increased 14C-PGD2 by 13-19% in both cell lines and increased 14C-PGE2 by 20% in the SW480 cell line only. LA significantly (P < 0.05) increased 5-hydroperoxyeicosatetraenoate by 27% in the MCF-7 cell line. Lipid peroxidation, as determined by increased levels of 8-epi-prostaglandin F2alpha (8-epi-PGF2alpha), was observed following treatment with c9,t11-CLA isomer in both cell lines (P < 0.02) and with t10,c12-CLA isomer in the MCF-7 cell line only (P < 0.05). These data indicate that the growth-promoting effects of LA in the SW480 cell line may be associated with enhanced conversion of AA to PGE2 but that the growth-suppressing effects of CLA isomers in both cell lines may be due to changes in AA distribution among cellular lipids and an altered prostaglandin profile.

Arachidonic Acid↗

Alpha-eleostearic acid (9Z11E13E-18:3) is quickly converted to conjugated linoleic acid (9Z11E-18:2) in rats.

We previously showed that alpha-eleostearic acid (alpha-ESA; 9Z11E13E-18:3) is converted to conjugated linoleic acid (CLA; 9,11-18:2) in the liver and plasma of rats that were given diets including 1% alpha-ESA for 4 wk. In this study, we investigated this phenomenon in detail. First, the chemical structure of CLA produced by alpha-ESA administration was determined. After alpha-ESA was orally administered to rats, CLA in rat liver was isolated by HPLC. The positional and geometric isomerism was determined using GC-EI/MS and (13)C-NMR, respectively, and the CLA generated in rats after alpha-ESA feeding was confirmed to be 9Z11E-CLA. Next, the concentrations of alpha-ESA and CLA were determined 0, 3, 6, and 24 h after oral administration of alpha-ESA to rats. Moreover, we also investigated whether enteric bacteria are involved in the conversion of alpha-ESA to CLA using germ-free rats. alpha-ESA was orally administered to germ-free and normal rats and alpha-ESA and CLA were detected in the organs of both groups. In addition, to confirm that this reaction was enzyme-mediated, alpha-ESA was reacted with tissue homogenates (liver, kidney, and small intestine mucous) and coenzymes (NADH, NAD(+), NADPH, and NADP(+)), and the enzyme activities were estimated from the amount of CLA produced. CLA was detected when alpha-ESA was reacted with liver, kidney, and small intestine mucous homogenates and a coenzyme (NADPH). These results indicated that alpha-ESA is converted to 9Z11E-CLA in rats by a Delta13-saturation reaction carried out by an NADPH-dependent enzyme.

Animals↗

The effect of dietary conjugated linoleic acid on egg yolk fatty acids and hatchability in Japanese quail.

Conjugated linoleic acid (CLA) increased the ratio of saturated fatty acids to monounsaturated fatty acids in yolk and caused embryo mortality. Our preliminary studies showed that CLA had less of an effect on hatchability of quail than chickens. Hence, the objective was to determine the effects of dietary CLA on quail egg fatty acid content and hatchability. Eight male-female Japanese quail pairs per group were randomly assigned to diets containing 0 (canola oil; CO), 0.25, 0.5, 1, 2, or 3% CLA for 8 wk. Eggs were collected, held at 15 degrees C for 24 h, and then incubated. Three eggs from each group were collected for fatty acid analysis on the 45th day. At the end of the 8 wk, all quail were euthanized. Liver samples from female quail were obtained for fatty acid analysis. Diet containing 3, 2, or 1% CLA caused 100% embryo mortality after 6, 10, or 12 d of feeding, whereas overall hatchabilities in groups 0, 0.25, and 0.5 were 84, 86, and 64%, respectively. As the dietary CLA increased, egg and hepatic CLA increased, C16:0 increased and C16:1(n-7) and C18:1(n-9) decreased, whereas C18:0 remained unchanged. Diets containing 1, 2, or 3% CLA decreased the C20:4(n-6) levels in yolk (significantly) and liver (inconsistently) lipids. Yolk CLA levels from 0, 0.25, 0.5, 1, 2, and 3% CLA were 0.31, 0.90, 1.48, 2.44, 5.88, and 11.2%, respectively. The ratios of C16:0/C16:1(n-7) in yolks from groups fed 0, 0.25, 0.5, 1, 2, or 3% CLA were 8.2, 16.3, 20.4, 24.6, 26.1, and 28.6, respectively. The ratios of C18:0/C18:1(n-9) in yolks from hens fed 0, 0.25, 0.5, 1, 2, or 3% CLA were 0.28, 0.40, 0.48, 0.49, 0.69, and 0.83, respectively. Quail fed 0.25% CLA had increased egg size, whereas quail fed 2 or 3% had reduced egg size compared with those fed CO. Liver sizes (%) in all of the groups were increased, except for the group fed 0.25% CLA. These data suggest that CLA may affect hatchability possibly by changing the fatty acid composition of the yolk.

Animal Feed↗

Conjugated linoleic acid isomers and their conjugated derivatives inhibit growth of human cancer cell lines.

Conjugated linoleic acid (CLA), mainly c9,t11- and t10,c12-isomers, and polyunsaturated n-3 fatty acids (n-3 PUFA) have been shown to reduce tumor growth. This study compared, on a set of human tumor cells (breast, lung, colon, prostate and melanoma), the antiproliferative effects of: i) trans monounsaturated fatty acids (MUFA) vs. cis MUFA and MUFA vs. PUFA, ii) individual isomers of CLA vs. linoleic acid, iii) CLA-conjugated derivatives vs. their non-conjugated homologues and vs. CLA isomers. Tumor cells were exposed to medium containing individual FA (100 microM) for 48 h and their proliferation was determined by measuring the cellular DNA content (fluorescent Hoechst 33342 dye). The antiproliferative effects of FA varied with the type of cells and were mainly dependent on the degree of unsaturation and on the position and configuration of their double bonds. One isomer of CLA (t9,t11-18:2) and CLA-conjugated derivatives exhibited the strongest growth-inhibitory effect against cancer cells. These results suggest that ruminant products contain active compounds against human tumor cell proliferation.

Cell Growth Processes↗

Photochemical production of conjugated linoleic acid from soybean oil.

Conjugated linoleic acid (CLA), an anticarcinogenic compound with numerous other health benefits, is present mainly in dairy and beef lipids. The main CLA isomer present in dairy and beef lipids is cis 9, trans 11 CLA at a 0.5% concentration. The typical minimum human dietary intake of CLA is 10 times less than the 3 g/d suggested requirement that has been extrapolated from animal and cell-line studies. The objectives of this study were to produce CLA isomers from soybean oil by photoisomerization of soybean oil linoleic acid and to study the oxidation status of the oil. Refined, bleached, and deodorized soybean oil with added iodine concentrations of 0, 0.1, 0.25, and 0.5% was exposed to a 100-W mercury lamp for 0 to 120 h. An SP-2560 fused-silica capillary GC column with FID was used to analyze the esterified CLA isomers in the photoisomerized oil. The CLA content of the individual isomers was optimized by response surface methodology. Attenuated total reflectance (ATR)-FTIR spectra in the 3400 to 3600 cm(-1) range and 1H NMR spectra in the 8 to 12 ppm range of the photoisomerized soybean oil were obtained to follow hydroperoxide formation. The largest amount of cis 9, trans 11 CLA isomer in soybean oil was 0.6%, obtained with 0.25% iodine and 84 h of photoisomerization. Lipid hydroperoxide peaks in the ATR-FTIR spectra and aldehyde peaks in the 1H NMR spectra were not observed in the photoisomerized soybean oil, and the spectra were similar to that of fresh soybean oil. This study shows that CLA isomers can be produced simply and inexpensively from soybean oil by photoisomerization.

Hydrogen Peroxide↗

Antiatherogenic effects of structured lipid containing conjugated linoleic acid in C57BL/6J mice.

Conjugated linoleic acids (CLA) were enzymatically acidolyzed with olive oil to produce structured lipids (SL), and their antiatherosclerotic properties were investigated in C57BL/6J mice. Twenty-eight mice were divided into four groups and fed control diet or atherogenic diets supplemented with high cholesterol and high fat (HCHF) containing 5% of lard, olive oil, or SL based on control diet for 4 weeks. The supplementation of SL diet (0.6% CLA) significantly reduced the levels of serum total cholesterol and total triglyceride and increased high-density lipoprotein cholesterol level as compared to lard and olive oil diet groups (p < 0.05). The activity of liver acyl CoA:cholesterol acyltransferase (ACAT) of mice fed the SL diet was significantly lower than that of mice fed the lard or olive oil diet. A reduced formation of aortic fatty streak was observed in SL group. The extent of CLA incorporation depended on tissues or types of phospholipids. More CLA was incorporated in adipose tissue (1.85 mol %) than in the liver (0.33 mol %). Besides, more CLA was found in phosphatidylethanolamine (PE) (0.47 mol %) than in phosphatidylcholine (PC) (0.05 mol %) of hepatic phospholipids. Hepatic phospholipids (PC and PE) of mice fed the SL diet contained reduced contents of arachidonic and linoleic acid compared with mice fed the olive oil or lard diet. The present study suggests that SL could be considered as a functional oil for preventing risks of atheroscelerosis.

Animals↗

Mechanisms of action of conjugated linoleic acid: evidence and speculation.

Conjugated linoleic acid (CLA) has been shown to inhibit carcinogenesis and atherosclerosis, enhance immunologic function while protecting against the catabolic effects of immune stimulation, affect body composition change (reducing body fat gain while enhancing lean body mass gain), and stimulate the growth of young rats. We discuss possible biochemical mechanisms that underlie these physiological effects. We emphasize the importance of considering the effects, both individually and combined, of the two CLA isomers (cis-9, trans-11 CLA and trans-10, cis-12 CLA) that have been shown to exhibit biological activity and which appear to exert their effects via different biochemical mechanisms.

Animals↗

Dose response of milk fat to intravenous administration of the trans-10, cis-12 isomer of conjugated linoleic acid.

Intravenous infusion of conjugated linoleic acid (CLA) was evaluated as a simpler method than abomasal infusion and the feeding of calcium salts to examine milk fat depression. The objectives were to determine the dose-dependent response of milk fat and plasma metabolites to intravenous administration of the trans-10, cis-12 isomer of CLA, an isomer identified to possess an inhibitory effect on milk fat synthesis. Four multiparous Holstein cows averaging 123 +/- 30 d in milk were randomly assigned to treatments in a 4 x 4 Latin square design. Catheters were inserted into the jugular vein for infusions and blood sampling. Treatments consisted of intravenous infusions of 0, 2, 4, and 6 g/d CLA (> 95% trans-10, cis-12 CLA). Infusates contained 72 g/d of a parenteral solution, saline, and CLA to 90 ml. Periods were of 5 d duration with a 7 d wash out. Milk was sampled at each milking and analyzed for fat, protein, and fatty acids. Blood samples were obtained on the last day of each period. Dry matter intake (22.4 +/- 2.4 kg/d), milk yield (28.5 +/- 3.3 kg/d), and protein percent (3.26 +/- 0.08%) of cows were not affected by treatment. However, milk fat percentage was reduced linearly with increasing doses of CLA. Milk fat percentage was 4.17, 3.53, 3.29, and 2.92% on d 5 for treatments 0, 2, 4, and 6 g/d CLA, respectively. Concentrations (4.2 mg/g of fat) of cis-9, trans-11 CLA in milk fat were not affected by treatment. However, an increase in the trans-10, cis-12 CLA content of milk fat was observed. Milk fat contained 0.00, 0.02, 0.06, and 0.10 mg of trans-10, cis-12 CLA per g of fat (SEM = 0.065) for treatments 0, 2, 4, and 6 g/d CLA, respectively. Plasma NEFA concentration increased linearly with the dose of the trans-10, cis-12 CLA. Intravenous infusion of the trans-10, cis-12 isomer of CLA depressed milk fat in a linear manner over the range of infusion studied and, therefore, is an alternative to abomasal infusion.

Abomasum↗

Dietary conjugated linoleic acid and body composition.

Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of conjugated dienoic derivatives of linoleic acid. The major dietary source of CLA for humans is ruminant meats, such as beef and lamb, and dairy products, such as milk and cheese. The major isomer of CLA in natural food is cis-9,trans-11 (c9,t11). The commercial preparations contain approximately equal amounts of c9,t11 and trans-10,cis-12 (t10,c12) isomers. Studies have shown that CLA, specifically the t10,c12-isomer, can reduce fat tissue deposition and body lipid content but appears to induce insulin resistance and fatty liver and spleen in various animals. A few human studies suggest that CLA supplementation has no effect on body weight and could reduce body fat to a much lesser extent than in animals. To draw conclusions on this form of dietary supplementation and to ultimately make appropriate recommendations, further human studies are required. The postulated antiobesity mechanisms of CLA include decreased energy and food intakes, decreased lipogenesis, and increased energy expenditure, lipolysis, and fat oxidation. This review addresses recent studies of the effects of CLA on lipid metabolism, fat deposition, and body composition in both animals and humans as well as the mechanisms surrounding these effects.

Adipose Tissue↗

Modification of skin composition by conjugated linoleic acid alone or with combination of other fatty acids in mice.

The effects of conjugated linoleic acid (CLA), gamma-linolenic acid (GLA), linoleic acid (LA), and their combinations, on skin composition in mice were investigated. Mice (8 weeks old) were orally administered with either LA, GLA, CLA, LA + GLA, LA + CLA, or CLA + GLA for 4 weeks. Then, the skin was analysed for triacylglycerol content, fatty acid composition and collagen content. Additionally, thicknesses of the dermis layer and subcutaneous tissue layer, and the size and number of adipocytes were measured histologically. The skin fatty acid composition was modified depending upon the fatty acid composition of supplemented oils. In each oil-alone group, skin triacylglycerol content was the highest in LA, followed by GLA and CLA treatments. Combinations with CLA had a similar triacylglycerol content compared with the CLA-alone group. No significant changes in collagen content were observed among any treatments. The effects on subcutaneous thickness were similar to the results obtained in the triacylglycerol contents, where groups supplemented with CLA alone or other fatty acids had significantly thinner subcutaneous tissue compared with the LA-alone group. However, no significant difference was detected in the thickness of the dermis layers. The number of adipocytes was highest in the LA + GLA group and tended to be reduced by CLA with or without the other fatty acids. These results suggest that CLA alone or in combination with other fatty acids strongly modifies skin composition in mice.

Administration, Oral↗

Linoleic acid conjugation by human intestinal microorganisms is inhibited by glucose and other substrates in vitro and in gnotobiotic rats.

The anticarcinogen conjugated linoleic acid (CLA) is a product of bacterial activity that isomerizes linoleic acid (LA) in the rumen of herbivores. Therefore, fatty dairy products in the human diet are enriched with CLA. Although bacteria capable of in vitro LA conjugation were detected in the human intestinal tract, CLA synthesis from dietary sunflower seed oil was not observed in gnotobiotic rats associated with these intestinal bacteria. The objective of the study was to investigate variables that affect LA conjugation. In vitro, LA conjugation was strongly inhibited by glucose and other substrates. Concentrations of 1.5 mmol glucose/L inhibited LA conjugation by 50%. Methyl-alpha-D-glucoside was a less effective inhibitor than glucose, and 2-deoxy-D-glucose did not inhibit LA conjugation at all. To analyze the concentration of carbohydrates in intestinal contents, the LA-conjugating bacterial mixed culture and human fecal microorganisms were introduced into germ-free rats. Samples of feces and cecum and colon contents of both groups exhibited in vitro LA-conjugating activity. Rats associated with human intestinal microorganisms contained 5.7 +/- 1. 3 mmol glucose/L in the cecal contents and 6.6 +/- 1.0 mmol glucose/L in the colonic contents. Rats associated with CLA-producing bacterial culture contained 3.4 +/- 1.3 mmol glucose/L in the cecal contents and 4.2 +/- 1.0 mmol glucose/L in the colonic contents. These values are within a range that may explain the observed inhibition of LA conjugation in vivo.

Animals↗

Trans-vaccenic acid is desaturated to conjugated linoleic acid in mice.

Mice were fed pure trans11 octadecenoic acid (trans-vaccenic acid; TVA) to determine whether it is desaturated to cis9, trans11 octadecadienoic acid, a predominant isomer of conjugated linoleic acid (CLA). In a preliminary trial, 12% of the TVA consumed during a 2-wk feeding period was recovered in the carcass as CLA. As a proportion of TVA in the tissues available for bioconversion, 48.8% was desaturated. We tested whether desaturation could be modified by supplementing no modifier, 0.5% clofibric acid to stimulate desaturation, or increasing the polyunsaturated fatty acids (PUFA) (10% corn oil vs. 4% corn oil) to inhibit desaturation in diets with or without 1% TVA. These diets were fed to six groups of mice in a 3x2 factorial arrangement of treatments. Feeding 1% TVA with 10% corn oil decreased feed intake (2.70 vs. 3.73 g/d, SEM 0.23; P<0.05). Bioconversion of dietary TVA was 12.0, 7.5 and 5.1% for mice fed no modifier of desaturation, clofibrate and increased PUFA, respectively. Conversion based on TVA available for desaturation was 52.6, 55.5 and 37.0%, respectively. Thus, clofibrate did not increase bioconversion, but increasing PUFA decreased conversion by 30%. To test whether TVA decreases food intake directly or after conversion to CLA, four groups of mice were fed diets containing 1% stearic, TVA, elaidic or conjugated linoleic acid. Dietary CLA decreased food intake and body fat, but did not change body protein. CLA was found in the carcass only when TVA or CLA was fed. CLA was found in both triacylglycerol and phospholipids when CLA was fed, but only in triacylglycerol when TVA was fed, suggesting that bioconversion occurred in the adipose tissue. In three trials, conversion of dietary TVA to CLA was 11.4+/-1.25%; conversion of stored TVA was 50.8+/-1.91%. Similar bioconversion of TVA in humans would increase current estimates of CLA available for the general population by 6- to 10-fold.

Administration, Oral↗

Conjugated linolenic acid is slowly absorbed in rat intestine, but quickly converted to conjugated linoleic acid.

We showed previously that alpha-eleostearic acid (alpha-ESA; 9Z11E13E-18:3) is converted to 9Z11E-conjugated linoleic acid (CLA) in rats through a Delta13-saturation reaction. To investigate this further, we examined the absorption and metabolism of alpha-ESA in rat intestine using a lipid absorption assay in lymph from the thoracic duct. In this study, we used 4 test oils [tung oil, perilla oil, CLA-triacylglycerol (TG), and pomegranate seed oil, containing alpha-ESA, alpha-linolenic acid (LnA; 9Z12Z15Z-18:3), CLA, and punicic acid (PA; 9Z11E13Z-18:3), respectively]. Emulsions containing the test oils were administered to rats, and lymph from the thoracic duct was collected over 24 h. The positional and geometrical isomerism of CLA produced by PA metabolism was determined using GC-electron impact (EI)-MS and (13)C-NMR, respectively; the product was confirmed to be 9Z11E-CLA. A part of alpha-ESA and PA was converted to 9Z11E-CLA 1 h after administration; therefore the lymphatic recoveries of alpha-ESA and PA were modified by the amount of recovered CLA. Cumulative recovery of CLA, alpha-ESA, and PA was lower than that of LnA only during h 1 (P < 0.05), and cumulative recovery of alpha-ESA and PA was significantly lower than that of LnA and CLA for 8 h (P < 0.05). Therefore, the absorption rate was LnA > CLA > alpha-ESA = PA. The conversion ratio of alpha-ESA to 9Z11E-CLA was higher than that of PA to 9Z11E-CLA over 24 h (P < 0.05). These results indicated that alpha-ESA and PA are slowly absorbed in rat intestine, and a portion of these fatty acids is quickly converted to 9Z11E-CLA.

Animals↗