Human studies using isotope labeled fatty acids: answered and unanswered questions.

Human studies using deuterium-labeled fatty acids have answered many questions related to the metabolism and health effects of dietary fats. These studies also raised a number of unanswered questions and unresolved issues. For example, studies with cis and trans positional isomers dispelled concerns and allegations that the isomers in partially hydrogenated fats were poorly absorbed, accumulate in undesirable phospholipid acyl positions, mimic stearic acid and competed with oleic acid.

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.

Stearic acid absorption and its metabolizable energy value are minimally lower than those of other fatty acids in healthy men fed mixed diets.

Compared with other saturated fatty acids, stearic acid appears to have different metabolic effects with respect to its impact on risk for cardiovascular disease. These differences may in part reflect biologically important differences in absorption. This study was designed to compare the absorption and the metabolizable energy value of stearic acid with other fatty acids from mixed diets fed to healthy humans.

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.

Stable isotope approaches, applications, and issues related to polyunsaturated fatty acid metabolism studies.

The use of stable isotope tracers for investigating fatty acid metabolism in human subjects has increased substantially over the last decade. Advances in analytical instrumentation, commercial availability of labeled substrates, and safety considerations are major reasons for this increased use of stable isotope tracers. Several experimental design options are available for using either deuterium or carbon-13 as tracers for fatty acid and lipid 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.

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.

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.

trans Fatty acids: infant and fetal development.

This review evaluates scientific data associated with the possibility that trans fatty acids compromise fetal and infant early development. Concerns have been triggered by research that has heightened our awareness of the importance of n-3 and n-6 fatty acids; shown that trans fatty acids inhibit delta6 desaturation of linoleic acid; identified trans fatty acid isomers in fetal, infant, and maternal tissues; and reported an inverse association between the trans fatty acid content of tissue lipids and measures of growth and development. Animal studies provide little evidence that trans fatty acids influence growth, reproduction, or gross aspects of fetal development.

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.

A human dietary arachidonic acid supplementation study conducted in a metabolic research unit: rationale and design.

While there are many reports of studies that fed arachidonic acid (AA) to animals, there are very few reports of AA feeding to humans under controlled conditions. This 130-d study was conceived as a controlled, symmetrical crossover design with healthy, adult male volunteers. They lived in the metabolic research unit (MRU) of the Western Human Nutrition Research (WHNRC) for the entire study.

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).

Delta 8 desaturation in vivo of deuterated eicosatrienoic acid by mouse liver.

In vitro evidence has been reported for an alternate pathway that involves delta 8 desaturation of n-6 and n-3 polyunsaturated fatty acids (PUFA). The present study was designed to allow detection of delta 8 desaturation in vivo and to provide an estimation of the relative contribution of delta 8 desaturation to the in vivo synthesis of n-3 fatty acids. Male adult ICR mice were fed a semisynthetic fat-free diet for eight days, and then the diets were supplemented for three days with deuterated 11,14,17-eicosatrienoic acid (20:3-d8) labeled at the 3,3,4,4,8,8,9,9 carbon positions.

Analysis of triglycerides using atmospheric pressure chemical ionization mass spectrometry.

Atmospheric pressure chemical ionization (APCI) mass spectrometry was investigated as a new method for analysis of a mixture of triglycerides separated by reverse-phase high-performance liquid chromatography. A mixture of homogeneous (monoacid) triglyceride standards containing fatty acids with zero to three double bonds was analyzed to demonstrate the quality of mass spectra obtained by using the APCI interface. The mass spectra showed that minimal fragmentation occurs, resulting primarily in diglyceride [M-RCOO]+ ions and [M + 1]+ protonated molecular ions.

Metabolism of dietary stearic acid relative to other fatty acids in human subjects.

This paper reviews results obtained by stable-isotope-tracer methods for stearic acid (18:0) and palmitic acid (16:0) metabolism and the influence of dietary linoleic acid on the metabolism of these saturated fatty acids in humans subjects. The results, based on stable-isotope-tracer data, show that absorption of 18:0 is not significantly different from 16:0; percent desaturation of 18:0 to 9-cis 18:1 (9.2%) is 2.

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.

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.

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.

Alternate pathways in the desaturation and chain elongation of linolenic acid, 18:3(n-3), in cultured glioma cells.

Cultured C6 glioma cells rapidly incorporate and metabolize the essential fatty acids, 18:2(n-6) and 18:3(n-3), to 20- and 22-carbon polyunsaturated fatty acids. Using several deuterated fatty acid substrates we have obtained data that suggest alternate pathways, one possibly involving delta 8-desaturation, may exist in glioma cells for formation of 20:5(n-3) and 22:6(n-3) from 18:3(n-3). With 18:3(n-3)-6,6,7,7-d4 practically no 18:4(n-3)-6,7-d2 or 20:4(n-3)-8,9-d2 was detected whereas 20:3(n-3)-8,8,9,9-d4 accounted for 3.

Metabolites of cis,trans, and trans,cis isomers of linoleic acid in mice and incorporation into tissue lipids.

Metabolism of octadecadienoic acid isomers in weanling mice was studied by feeding fat-free diets supplemented with 2% by weight of cis-9,trans-12-octadecadienoic acid (c,t-18:2-d0), tetradeuterated trans-9,cis-12-octadecadienoic acid (t,c-18:2-d4) or dideuterated cis-9,cis-12-octadecadienoic acid (c,c-18:2-d2). Rates for conversion of c,t-18:2-d0 and c,c-18:2-d2 to c,t-20:4-d0 and c,c-20:4-d2 were identical and both were 5-times higher than conversion of t,c-18:2-d4 to t,c-20:4-d4. Accumulation of t,c-18:2-d4 in liver lipids was 2-4-times higher than for c,t-18:2-d0 or c,c-18:2-d2.

Correlation of isomeric fatty acids in human adipose tissue with clinical risk factors for cardiovascular disease.

The relationships between the adipose tissue concentrations of 19 geometric and positional fatty acid isomers and 10 cardiovascular disease risk factors were determined in 76 free-living adult males. The percentages for trans isomers (total mean +/- SD 4.14 +/- 0.

Measurement of the metabolic interconversion of deuterium-labeled fatty acids by gas chromatography/mass spectrometry.

An analytical method that was developed to analyze deuterium-labeled fatty acids in human blood has been extended to identify labeled fatty acids from C14 to C24 chain length which are formed by metabolic processes such as desaturation, elongation, or shortening of the labeled fatty acids fed. A new computer and a hardware adder have been utilized to assure reliable data acquisition. Relative standard deviations for the analysis of labeled fatty acids were measured at 0.