Effect of triacylglycerol structure on absorption and metabolism of isotope-labeled palmitic and linoleic acids by humans.

The effect of dietary TAG structure and fatty acid acyl TAG position on palmitic and linoleic acid metabolism was investigated in four middle-aged male subjects. The study design consisted of feeding diets containing 61 g/d of native lard (NL) or randomized lard (RL) for 28 d. Subjects then received an oral dose of either 1,3-tetradeuteriopalmitoyl-2-dideuteriolinoleoyl-rac-glycerol or a mixture of 1,3-dideuteriolinoleoyl-2-tetradeuteriopalmitoyl-rac-glycerol and 1,3-hexadeuteriopalmitoyl-2-tetradeuteriolinoleoyl-rac-glycerol. Methyl esters of plasma lipids isolated from blood samples drawn over a 2-d period were analyzed by GC-MS. Results showed that absorption of the 2H-fatty acids (2H-FA) was not influenced by TAG position. The 2H-FA at the 2-acyl TAG position were 85+/-4.6% retained after absorption. Substantial migration of 2H-16:0 (31.2+/-8.6%) from the sn-2 TAG position to the sn-1,3 position and 2H-18:2n-6 (52.8+/-6.4%) from the sn-1,3 position to the sn-2 position of chylomicron TAG occurred after initial absorption and indicates the presence of a previously unrecognized isomerization mechanism. Incorporation and turnover of the 2H-FA in chylomicron TAG, plasma TAG, and plasma cholesterol esters were not influenced by TAG acyl position. Accretion of 2H-16:0 from the sn-2 TAG position in 1-acylphosphatidylcholine was 1.7 times higher than 2H-16:0 from the sn-1,3 TAG positions. Acyl TAG position did not influence 2H-18:2n-6 incorporation in PC. The concentration of 2H-18:2n-6-derived 2H-20:4n-6 in plasma PC from subjects fed the RL diet was 1.5 times higher than for subjects fed the NL diet, and this result suggests that diets containing 16:0 located at the sn-2 TAG position may inhibit 20:4n-6 synthesis. The overall conclusion is that selective rearrangement of chylomicron TAG structures diminishes but does not totally eliminate the metabolic and physiological effects of dietary TAG structure.

Separation of conjugated linoleic acid methyl esters by silver-ion high performance liquid chromatography in semi-preparative mode.

Silver-ion HPLC (Ag-HPLC) has been utilized to separate a variety of unsaturated fatty acid methyl esters (FAMEs) by configuration, location or number of olefinic or acetylenic bonds. Two analytical Ag-HPLC columns connected in series and an isocratic solvent system of acetonitrile (ACN) in hexane were used to fractionate 10-15 mg samples of a mixture of two deuterium-labeled isomers of conjugated linoleic acid (Z9.E11- and Z9,Z11-octadecadienoic acid-17,17,18,18-d4). "Baseline" (> 95%) resolution of the two isomers, which decreased with increasing weights of sample injected, was maintained by careful adjustment of the percentage of ACN in the ACN/hexane solvent system. Chemical purities of the isolated FAME were > 96%.

Effect of dietary conjugated linoleic acid (CLA) on metabolism of isotope-labeled oleic, linoleic, and CLA isomers in women.

The purpose of this study was to investigate the effect of dietary CLA on accretion of 9c-18:1, 9c,12c-18:2, 10t,12c-18:2, and 9c,11t-18:2 and conversion of these FA to their desaturated, elongated, and chain-shortened metabolites. The subjects were six healthy adult women who had consumed normal diets supplemented with 6 g/d of sunflower oil or 3.9 g/d of CLA for 63 d. A mixture of 10t,2c-18:2-d4, 9c,11t-18:2-d6, 9c-18:1-d8, and 9c,12c-18:2-d2, as their ethyl esters, was fed to each subject, and nine blood samples were drawn over a 48-h period. The results show that dietary CLA supplementation had no effect on the metabolism of the deuterium-labeled FA. These metabolic results were consistent with the general lack of a CLA diet effect on a variety of physiological responses previously reported for these women. The 2H-CLA isomers were metabolically different. The relative percent differences between the accumulation of 9c,11t-18:2-d6 and 10t,12c-18:2-d4 in plasma lipid classes ranged from 9 to 73%. The largest differences were a fourfold higher incorporation of 10t,12c-18:2-d4 than 9c,11t-18:2-d6 in 1-acyl PC and a two- to threefold higher incorporation of 9c,11t-18:2-d6 than 10t,12c-18:2-d4 in cholesterol esters. Compared to 9c-18:1-d8 and 9c,12c-18:2-d2, the 10t,12c-18:2-d4 and 9c,11t-18:2-d6 isomers were 20-25% less well absorbed. Relative to 9c-18:1, incorporation of the CLA isomers into 2-acyl PC and cholesterol ester was 39-84% lower and incorporation of 10t,12c-18:2 was 50% higher in 1-acyl PC. This pattern of selective incorporation and discrimination is similar to the pattern generally observed for trans and cis 18:1 positional isomers. Elongated and desaturated CLA metabolites were detected. The concentration of 6c,10t,12c-18:3-d4 in plasma TG was equal to 6.8% of the 10t,12c-18:2-d4 present, and TG was the only lipid fraction that contained a CLA metabolite present at concentrations sufficient for reliable quantification. In conclusion, no effect of dietary CLA was observed, absorption of CLA was less than that of 9c-18:1, CLA positional isomers were metabolically different, and conversion of CLA isomers to desaturated and elongated metabolites was low.

Analysis of conjugated linoleic acid-enriched triacylglycerol mixtures by isocratic silver-ion high-performance liquid chromatography.

Silver-ion HPLC (Ag-HPLC) was applied to the fractionation of a triacylglycerol (TAG) sample enriched (>80%) with conjugated linoleic acid (CLA). After conversion of the TAGs to fatty acid methyl esters using sodium methoxide in methanol, Ag-HPLC (dual-column; isocratic solvent system of 0.1% acetonitrile in hexane; UV detection at 233 nm) was used to determine the CLA isomer distribution (50:50 mixture of 9c 11t- and 10t,12c-18:2). Three or four Ag-HPLC columns connected in series (0.6-1.0% acetonitrile in hexane as solvent; UV detection at 206 nm) were used to analyze the sample in TAG form. Elution times for CLA-enriched TAGs averaged 30 min or less. Isocratic solvent conditions were used to eliminate the solvent equilibration times (often 30 min or more) required between sample injections when solvent programming is used. The ratio of TAGs containing three vs. only two CLA molecules was found to be approximately 3 to 1. Ag-HPLC has thus been shown to be a useful method for rapidly analyzing not only CLA isomers as esters, but also in the TAG form.

Changes in conjugated linoleic acid composition within samples obtained from a single source.

Conjugated linoleic acid (CLA; 9c,11t-18:2) and CLA isomers have been reported, in animals, to exhibit a variety of health-related benefits. Silver ion high-performance liquid chromatography (Ag-HPLC) was found to provide better resolution of the isomers than gas chromatography. Most commercially available samples of CLA, prepared by base-catalyzed isomerization of linoleic acid (9c,12c-18:2), are composed of mixtures of four major isomers. While these isomers have been characterized, we found significant changes in CLA isomer rations within samples obtained from the same producer/commercial supplier over a period of 1.5 yr. In the first sample, the four cis/trans isomers (8t,10c-18:2, 9c,11t-18:2, 10t,12c-18:2 and 11c,13t-18:2) were present in a ratio of approximately 1:2:2:1, while in the second sample they were present in almost equal proportions. If indeed certain daily levels of CLA intake are required to produce suggested health benefits in humans, changes in concentrations of specific CLA isomers could significantly impact these effects. Care must be taken to analyze the CLA used in human and animal studies.

Biosynthesis of conjugated linoleic acid in humans.

This paper deals with the reanalysis of serum lipids from previous studies in which deuterated fatty acids were administered to a single person. Samples were reanalyzed to determine if the deuterated fatty acids were converted to deuterium-labeled conjugated linoleic acid (CLA, 9c,11t-18:2) or other CLA isomers. We found 11-trans-octadecenoate (fed as the triglyceride) was converted (delta9 desaturase) to CLA, at a CLA enrichment of ca. 30%. The 11-cis-octadecenoate isomer was also converted to 9c,11c-18:2, but at <10% the concentration of the 11t-18:1 isomer. No evidence (within our limits of detection) for conversion of 10-cis- or 10-trans-octadecenoate to the 10,12-CLA isomers (delta12 desaturase) was found. No evidence for the conversion of 9-cis,12-cis-octadecadienoate to CLA (via isomerase enzyme) was found. Although these data come from four single human subject studies, data from some 30 similar human studies have convinced us that the existence of a metabolic pathway in one subject may be extrapolated to the normal adult population.

Effect of dietary docosahexaenoic acid on desaturation and uptake in vivo of isotope-labeled oleic, linoleic, and linolenic acids by male subjects.

The effect of dietary docosahexaenoic acid (22:6n-3, DHA) on the metabolism of oleic, linoleic, and linolenic acids was investigated in male subjects (n = 6) confined to a metabolic unit and fed diets containing 6.5 or <0.1 g/d of DHA for 90 d. At the end of the diet period, the subjects were fed a mixture of deuterated triglycerides containing 18:1n-9[d6], 18:2n-6[d2], and 18:3n-3[d4]. Blood samples were drawn at 0, 2, 4, 6, 8, 12, 24, 48, and 72 h. Methyl esters of plasma total lipids, triglycerides, phospholipids, and cholesterol esters were analyzed by gas chromatography-mass spectrometry. Chylomicron triglyceride results show that the deuterated fatty acids were equally well absorbed and diet did not influence absorption. Compared to the low-DHA diet (LO-DHA), clearance of the labeled fatty acids from chylomicron triglycerides was modestly higher for subjects fed the high DHA diet (HI-DHA). DHA supplementation significantly reduced the concentrations of most n-6[d2] and n-3[d4] long-chain fatty acid (LCFA) metabolites in plasma lipids. Accumulation of 20:5n-3[d4] and 22:6n-3[d4] was depressed by 76 and 88%, respectively. Accumulations of 20:3n-6[d2] and 20:4n-6[d2] were both decreased by 72%. No effect of diet was observed on acyltransferase selectivity or on uptake and clearance of 18:1n-9[d6], 18:2n-6[d2], and 18:3n-3[d4]. The results indicate that accumulation of n-3 LCFA metabolites synthesized from 18:3n-3 in typical U.S. diets would be reduced from about 120 to 30 mg/d by supplementation with 6.5 g/d of DHA. Accumulation of n-6 LCFA metabolites synthesized from 18:2n-6 in U.S. diets is estimated to be reduced from about 800 to 180 mg/d. This decrease is two to three times the amount of n-6 LCFA in a typical U.S. diet. These results support the hypothesis that health benefits associated with DHA supplementation are the combined result of reduced accretion of n-6 LCFA metabolites and an increase in n-3 LCFA levels in tissue lipids.

Preparation, separation, and confirmation of the eight geometrical cis/trans conjugated linoleic acid isomers 8,10- through 11,13-18:2.

Conjugated linoleic acid (CLA) mixtures were isomerized with p-toluenesulfinic acid or I2 catalyst. The resultant mixtures of the eight cis/trans geometric isomers of 8,10-, 9,11-, 10,12-, and 11,13-octadecadienoic (18:2) acid methyl esters were separated by silver ion-high-performance liquid chromatography (Ag+-HPLC) and gas chromatography (GC). Ag+-HPLC allowed the separation of all positional CLA isomers and geometric cis/trans CLA isomers except 10,12-18:2. However, one of the 8,10 isomers (8cis, 10trans-18:2) coeluted with the 9trans,11cis-18:2 isomer. There were differences in the elution order of the pairs of geometric CLA isomers resolved by Ag+-HPLC. For the 8,10 and 9,11 CLA isomers, cis,trans eluted before trans,cis, whereas the opposite elution pattern was observed for the 11,13-18:2 geometric isomers (trans,cis before cis,trans). All eight cis/trans CLA isomers were separated by GC on long polar capillary columns only when their relative concentrations were about equal. Large differences in the relative concentration of the CLA isomers found in natural products obscured the resolution and identification of a number of minor CLA isomers. In such cases, GC-mass spectrometry of the dimethyloxazoline derivatives was used to identify and confirm coeluting CLA isomers. For the same positional isomer, the cis,trans consistently eluted before the trans,cis CLA isomers by GC. High resolution mass spectrometry (MS) selected ion recording (SIR) of the molecular ions of the 18:1, 18:2, and 18:3 fatty acid methyl esters served as an independent and highly sensitive method to confirm CLA methyl ester peak assignments in GC chromatograms obtained from food samples by flame-ionization detection. The high-resolution MS data were used to correct for the nonselectivity of the flame-ionization detector.

Improved separation of conjugated fatty acid methyl esters by silver ion-high-performance liquid chromatography.

Operating from one to six silver ion-high-performance liquid chromatography (Ag+-HPLC) columns in series progressively improved the resolution of the methyl esters of conjugated linoleic acid (CLA) isomeric mixtures from natural and commercial products. In natural products, the 8 trans, 10 cis-octadecadienoic (18:2) acid was resolved from the more abundant 7 trans, 9 cis-18:2, and the 10 trans, 12 cis-18:2 was separated from the major 9 cis, 11 trans-18:2 peak. In addition, both 11 trans, 13 cis-18:2 and 11 cis, 13 trans-18:2 isomers were found in natural products and were separated; the presence of the latter, 11 cis, 13 trans-18:2, was established in commercial CLA preparations. Three Ag+-HPLC columns in series appeared to be the best compromise to obtain satisfactory resolution of most CLA isomers found in natural products. A single Ag+-HPLC column in series with one of several normal-phase columns did not improve the resolution of CLA isomers as compared to that of the former alone. The 20:2 conjugated fatty acid isomers 11 cis, 13 trans-20:2 and 12 trans, 14 cis-20:2, which were synthesized by alkali isomerization from 11 cis, 14 cis-20:2, eluted in the same region of the Ag+-HPLC chromatogram just before the corresponding geometric CLA isomers. Therefore, CLA isomers will require isolation based on chain length prior to Ag+-HPLC separation. The positions of conjugated double bonds in 20:2 and 18:2 isomers were established by gas chromatography-electron ionization mass spectrometry as their 4,4-dimethyloxazoline derivatives. The double-bond geometry was determined by gas chromatography-direct deposition-Fourier transform infrared spectroscopy and by the Ag+-HPLC relative elution order.

Effect of dietary arachidonic acid on metabolism of deuterated linoleic acid by adult male subjects.

The influence of dietary supplementation with 20:4n-6 on uptake and turnover of deuterium-labeled linoleic acid (18:2n-6[d2]) in human plasma lipids and the synthesis of desaturated and elongated n-6 fatty acids from 18:2n-6[d21 were investigated in six adult male subjects. The subjects were fed either a high-arachidonic acid (HIAA) diet containing 1.7 g/d or a low-AA (LOAA) diet containing 0.21 g/d of AA for 50 d. Each subject was then dosed with about 3.5 g of 18:2n-6[d2] as the triglyceride (TG) at 8:00 A.M., 12:00, and 5:00 P.M. The total 18:2n-6[d21] fed to each subject was about 10.4 g and is approximately equal to one-half of the daily intake of 18:2n-6 in a typical U.S. male diet. Nine blood samples were drawn over a 96-h period. Methyl esters of plasma total lipid (TL), TG, phospholipid, and cholesterol ester were analyzed by gas chromatography-mass spectroscopy. Dietary 20:4n-6 supplementation did not affect uptake of 18:2n-6[d2] in plasma lipid classes over the 4-d study period nor the estimated half-life of 24-36 h for 18:2n-6[d2]. The percentages of major deuterium-labeled desaturation and elongation products in plasma TL, as a percentage of total deuterated fatty acids, were 1.35 and 1.34% 18:3n-6[d2]; 0.53 and 0.50% 20:2n-6[d2]; 1.80 and 0.92% 20:3n-6[d2] and 3.13 and 1.51% 20:4n-6[d2] for the LOAA and HIAA diet groups, respectively. Trace amounts (<0.1%) of the 22:4n-6[d2] and 22:5n-6[d2] metabolites were present. Plasma TL concentration data for both 20:3n-6[d2] and 20:4n-6[d2] were 48% lower (P < 0.05) in samples from the HIAA diet group than in samples from the LOAA diet group. For a normal adult male consuming a typical U.S. diet, the estimated accumulation in plasma TL of 20:4n-6 synthesized from 20 g/d (68 mmole) of 18:2n-6 is 677 mg/d (2.13 mmole). Dietary supplementation with 1.5 g/d of 20:4n-6 reduced accumulation of 20:4n-6 synthesized from 20 g/d of 18:2n-6 to about 326 mg/d (1.03 mmole).

Influence of dietary arachidonic acid on metabolism in vivo of 8cis,11cis,14-eicosatrienoic acid in humans.

This study investigated the influence of dietary arachidonic acid (20:4n-6) on delta 5 desaturation and incorporation of deuterium-labeled 8cis,11 cis, 14-eicosatrienoic acid (20:3n-6) into human plasma lipids. Adult male subjects (n = 4) were fed diets containing either 1.7 g/d (HI20:4 diet) or 0.21 g/d (LO20:4 diet) of arachidonic acid for 50 d and then dosed with a mixture containing ethyl esters of 20:3n-6[d4] and 18:1n-9[d2]. A series of blood samples was sequentially drawn over a 72-h period, and methyl esters of plasma total lipid, triacylglycerol, phospholipids, and cholesteryl ester were analyzed by gas chromatography-mass spectrometry. Based on the concentration of 20:3n6[d4] in total plasma lipid, the estimated conversion of 20:3n-6[d4] to 20:4n-6[d4] was 17.7 +/- 0.79% (HI20:4 diet) and 2.13 +/- 1.44% (LO20:4 diet). The concentrations of 20:4n-6[d4] in total plasma lipids from subjects fed the HI20:4 and LO20:4 diets were 2.10 +/- 0.6 and 0.29 +/- 0.2 mumole/mL plasma/mmole of 20:3n-6[d4] fed/kg of body weight. These data indicate that conversion of 20:3n-6[d4] to 20:4n-6[d4] was stimulated 7-8-fold by the HI20:4 diet. Phospholipid acyltransferase was 2.5-fold more selective for 20:3n-6[d4] than 18:1n-9[d2], and lecithin:cholesterol acyltransferase was 2-fold more selective for 18:1n-9[ds] than 20:3n-6[d4]. These differences in selectivity were not significantly influenced by diet. Absorption of ethyl 20:3n-6[d4] was about 33% less than ethyl 18:1n-9[d2]. The sum of the n-6 retroconversion products from 20:3n-6[d4] in total plasma lipids was about 2% of the total deuterated fatty acids. Neither absorption nor retroconversion appears to be influenced by diet.

Quantitative analysis of triglycerides using atmospheric pressure chemical ionization-mass spectrometry.

Atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) was used for quantitative analysis of triglycerides (TG) separated by reverse-phase high-performance liquid chromatography. APCI-MS was used for analysis of mono-acid TG standards containing deuterated internal standard, of a synthetic mixture of heterogeneous TG, of randomized and normal soybean oils and of randomized and normal lard samples. Quantitation of the TG by four approaches based on APCI-MS were compared, and these were compared to quantitation obtained using liquid chromatography (LC) with flame-ionization detection (FID). The APCI-MS methods were based on (i) calibration curves from data for mono-acid TG standards, (ii) response factors obtained from a synthetic mixture of TG, (iii) response factors calculated from comparison of randomized samples to their statistically expected compositions, and (iv) response factors calculated from comparison of fatty acid (FA) compositions calculated from TG compositions to FA compositions obtained by calibrated gas chromatography (GC) with FID. Response factors derived from a synthetic mixture were not widely applicable to samples of disparate composition. The TG compositions obtained using APCI-MS data without application of response factors had average relative errors very similar to those obtained using LC-FID. Numerous TG species were identified using LC/APCI-MS which were undetected using LC-FID. Two quantitation methods, based on response factors calculated from randomized samples or on response factors calculated from FA compositions, both gave similar results for all samples. The TG compositions obtained using response factors calculated from FA compositions showed less average relative error than was obtained from LC-FID data, and were in good agreement with predicted compositions for the synthetic mixture and for randomized soybean oil and lard samples.

Dietary linoleic acid influences desaturation and acylation of deuterium-labeled linoleic and linolenic acids in young adult males.

The purpose of this study was to investigate the effect of dietary linoleic acid (18:2(n - 6)) on the conversion of 18:2(n - 6) and 18:3(n - 3) to their respective n - 6 and n - 3 metabolites; to compare the incorporation of these fatty acids into human plasma lipids; to evaluate the importance of dietary 18:3(n - 3) as a precursor for the biosynthesis of long-chain length n - 3 fatty acids. The approach used was to feed young adult male subjects (n = 7) diets containing 2 levels of linoleic acid (SAT diet, 15 g/day; PUFA diet, 30 g/day) for 12 days. A mixture of triacylglycerols containing deuterated linolenic (18:3(n - 3)) and linoleic (18:2(n - 6)) acids was fed and blood samples were drawn over a 48 h period. Concentrations of deuterated 18:3(n - 3) in plasma total lipid ranged from 309.2 to 606.4 microgram/ml and concentrations of 18:2(n - 6) ranged from 949.2 to 1743.3 micrograms/ml. The sum of the deuterated n - 3 long-chain length fatty acid metabolites in plasma total lipid were 116 +/- 4.3 micrograms/ml (SAT diet) and 41.6 +/- 12.4 micrograms/ml (PUFA diet). The total deuterated n - 6 fatty acid metabolites were 34.6 +/- 12.2 micrograms/ml (SAT diet) and 9.8 +/- 5.9 micrograms/ml (PUFA diet). The total percent conversion of deuterated 18:3(n - 3) to n - 3 fatty acid metabolites and deuterated 18:2(n - 6) to n - 6 fatty acid metabolites were 11-18.5% and 1.0-2.2%, respectively. The percentages for deuterated 20:5(n - 3), 22:5(n - 3) and 22:6(n - 3) (6.0%, 3.5%, and 3.8%) were much higher than for 20:3(n - 6) and 20:4(n - 6) (0.9% and 0.5%). Overall, conversion of deuterated 18:3(n - 3) and 18:2(n - 6) was reduced by 40-54% when dietary intake of 18:2(n - 6) was increased from 15 to 30 g/day. Comparison of the deuterated 18:3(n - 3) and 18:2(n - 6) data for plasma triacylglycerol and phosphatidylcholine (PC) indicated that 18:2(n - 6) was preferentially incorporated into PC. Dietary 18:2(n - 6) intake did not alter acyltransferase selectivity but activity was reduced when 18:2(n - 6) intake was increased. Based on these results, conversion of the 18:3(n - 3) in the US diet (2 g) is estimated to provide 75-85% of the long-chain length n - 3 fatty acids needed to meet daily requirements for some (but not all) adults.

Influence of linoleic acid on desaturation and uptake of deuterium-labeled palmitic and stearic acids in humans.

Objectives of this study were to investigate the desaturation of stearic acid (18:0) and palmitic acid (16:0), to determine if differences in their metabolism provide a reasonable explantation for differences in their effect on serum cholesterol levels, and to investigate the affect of linoleic acid on delta 9-desaturase products in man. Deuterium-labeled 16:0 and 18:0 were used to follow the metabolism of these fatty acids in young adult male subjects that were pre-fed diets containing two different levels of linoleic acid. Results indicate that absorption of 16:0 and 18:0 was similar when all components of the mixture used to formulate the deuterated fat mixture were kept above the melting point of tristearin. The percent of 18:0 desaturated to 9c-18:1 was higher than the percent of 16:0 desaturated to 9c-16:1 (9.2% vs. 3.9%). The subject-to-subject variability suggests that differences in ability to desaturate saturated fatty acids may be related to the variability observed in response of serum cholesterol levels to dietary saturated fatty acids. Data for the distribution of 16:0 and 18:0 between triacylglycerol and phosphatidylcholine (PC) was markedly different. Based on PC data, phospholipid acyltransferase selectivity was about 2-fold higher for 18:0 than for 16:0. A 2-fold difference in the linoleic acid content of the pre-fed diets had little influence on desaturation or distribution of 16:0 and 18:0 between plasma lipid classes. A deuterium isotope effect was estimated to reduce delta 9-desaturase enzyme activity by 30-50%.

Metabolism of meadowfoam oil fatty acids in mice.

Meadowfoam oil is unusual because over 95% of the fatty acids are 20- and 22-carbon aliphatic acids with cis double bonds located principally at the 5- and/or 13-position. Since little information is available on the metabolism of the 5c-20:1 and 5c,13c-22:2 fatty acids, an exploratory study in mice was conducted to investigate the metabolism of purified samples of the free fatty acids isolated from meadowfoam oil, and to determine the effect of meadowfoam oil on weight gain and tissue lipid composition. Mice fed diets containing 5% by wt of the purified 5c-20:1 or 5c,13c-22:2 for 6 days exhibited no apparent physiological problems. Total liver lipids from mice fed the purified fatty acid diets contained mean values of 2.0% 5c-20:1 and 2.1% 5c,13c-22:2; total heart lipids contained 1.7% 5c-20:1 and 10.7% 5c,13c-22:2. Liver total phospholipids from mice fed a 5% meadowfoam oil diet for 19 wk contained 1.4% 5c-20:1 and 1.9% 5c,13c-22:2. There was no evidence of desaturation, elongation or retroconversion. Weight gain for mice fed the meadowfoam oil diet for 19 wk was similar to mice fed corn oil, and was higher than for mice fed hydrogenated cottonseed oil. Considering the high 5c-20:1 and 5c,13c-22:2 content of the diets, the percentages of these fatty acids in mouse tissue lipids from both the short- and long-term studies were low. Weight gain was surprisingly good since the meadowfoam oil diet was essential fatty acid-deficient. Results of this initial investigation suggest that the 5c-20:1 and 5c,13c-22:2 fatty acids were utilized primarily for energy.(ABSTRACT TRUNCATED AT 250 WORDS)

Incorporation of deuterium-labeled fatty acids into human milk, plasma, and lipoprotein phospholipids and cholesteryl esters.

Fatty acid metabolism and the contribution of dietary fatty acids to milk cholesteryl ester (CE) and phospholipid (PL) were investigated in normal lactating mothers. The approach used was to feed mixtures of triglycerides containing deuterium-labeled palmitic acid (16:0-2H2), oleic acid (18:1-2H6), and linoleic acid (18:2-2H4). Milk and plasma samples were collected for 72 hr. Triglyceride (TG), CE, and PL fractions from milk, plasma, and lipoprotein were isolated and analyzed by gas-liquid chromatography and mass spectrometry. Data for the milk CE and PL fractions showed a definite selectivity for incorporation of 16:0-2H2 and 18:1-2H6 relative to 18:2-2H4. Based on the ratios of the deuterated fatty acids incorporated into the milk CE and PL samples, their incorporation times and isotopic enrichment data, it appears that these fatty acids are supplied mainly by the TG derived from chylomicrons and very low density lipoproteins. Plasma and lipoprotein CE data showed a progressive increase in 18:2-2H4 content, with 16:0-2H2 and 18:1-2H4 remaining relatively constant over the collection period. Plasma and lipoprotein PL data showed a higher rate for incorporation of 18:2-2H4 than 16:0-2H2 and 18:1-2H6 over the course of the sampling period. Comparison to previous data from adult males indicates lactation does not have a major effect on the general metabolism of these fatty acids. An increase with time in the isotopic enrichment of 18:2-2H4 in the plasma and lipoprotein CE and PL samples was observed which is consistent with in vitro selectivities reported for lecithin:cholesterol acyltransferase and phosphatidylcholine acyltransferase.(ABSTRACT TRUNCATED AT 250 WORDS)

Incorporation of trans-8- and cis-8-octadecenoic acid isomers in human plasma and lipoprotein lipids.

Mixtures of deuterium-labeled trans-8, cis-8 and cis-9-octadecenoic acids (8t-18:1, 8c-18:1, 9c-18:1) were fed as triglycerides (TG) to two adult male subjects. Blood samples were collected sequentially over a 48-hour period. Plasma and lipoprotein lipids were separated by thin layer chromatography and analyzed by gas chromatography-mass spectroscopy. Results indicate (i) absorption of the 8t- and 8c-18:1 isomers were similar to 9c-18:1; (ii) the 8t-18:1 isomer was cleared approximately 30% faster than 9c-18:1 from plasma TG; (iii) cholesterol ester samples contained 8.4 times less 8t-18:1 than 9c-18:1; (iv) incorporation at the 1-acyl phosphatidylcholine (PC) position was higher for 8t-18:1 and 8c-18:1 (2.2 and 1.7 times) than for 9c-18:1; and (v) discrimination at the 2-acyl PC position was 4.6-fold against 8t-18:1 and 1.3-fold against 8c-18:1 compared with 9c-18:1. Discrimination against uptake of the delta-8 isomers in both neutral and phospholipid classes suggests that both 8t- and 8c-18:1 may be preferentially oxidized relative to 9c-18:1. Except for triglycerides, data for each of the lipid classes from total plasma and individual lipoprotein samples were similar. These data indicate that differences for incorporation and turnover of the 8t- and 8c-18:1 isomers relative to 9c-18:1 are not substantially influenced by the lipoprotein classes. The maximum isotopic enrichment detected in the chylomicron triglycerides fractions was 60%, which indicates that a substantial amount of endogenous triglycerides was mobilized during absorption of the deuterated fats.