ABSTRACT
Two direct methylation procedures often used for the analysis of total fatty acids in biological samples were evaluated for their application to samples containing fatty amides. Methylation of 5 mg of oleamide (cis-9-octadecenamide) in a one-step (methanolic HCl for 2 h at 70 degrees C) or a two-step (sodium methoxide for 10 min at 50 degrees C followed by methanolic HCl for 10 min at 80 degrees C) procedure gave 59 and 16% conversions of oleamide to oleic acid, respectively. Oleic acid recovery from oleamide was increased to 100% when the incubation in methanolic HCl was lengthened to 16 h and increased to 103% when the incubation in methoxide was modified to 24 h at 100 degrees C. However, conversion of oleamide to oleic acid in an animal feed sample was incomplete for the modified (24 h) two-step procedure but complete for the modified (16 h) one-step procedure. Unsaturated fatty amides in feed and digesta samples can be converted to fatty acid methyl esters by incubation in methanolic HCl if the time of exposure to the acid catalyst is extended from 2 to 16 h.
Subject(s)
Animal Feed/analysis , Feces/chemistry , Oleic Acid/chemical synthesis , Oleic Acids/chemistry , Esters , Fatty Acids , Food Additives , Methylation , Temperature , Time FactorsABSTRACT
On the basis of earlier reports of reduced growth rate and fat accumulation in animals fed trans 18:1, a study was conducted to determine whether the effects of trans 18:1 on lipolysis and glucose utilization by adipocytes differed from effects of the cis isomer. Two experiments compared three fatty acid isomers (oleic, elaidic and vaccenic acids) at several concentrations and at several fatty acid to albumin ratios in cell media. Adipocytes were isolated from adipose tissue of rats by collagenase digestion and incubated for 2 h in media containing added fatty acids. Compared with oleic acid, both trans isomers reduced (P < 0.01) the amount of glucose converted to cell lipid in both experiments. Glucose oxidation to carbon dioxide also was lower for both trans fatty acids in Experiments 1 (P < 0.05) and 2 (P < 0.06). Lipolytic rates were increased (P < 0.01) in both experiments by replacing oleic acid with either of the trans isomers. Trans isomers of octadecenoic acid had catabolic effects on adipocyte metabolism that occurred regardless of the position of the double bond, the fatty acid concentration in media or the fatty acid to albumin ratio. These catabolic effects explain previous observations of reduced growth rate and fat accumulation when oleic acid in animal diets is replaced with a trans isomer.
Subject(s)
Adipocytes/metabolism , Glucose/metabolism , Lipolysis/drug effects , Oleic Acid , Oleic Acids/chemistry , Oleic Acids/pharmacology , Adipocytes/cytology , Adipocytes/drug effects , Albumins/metabolism , Animals , Cells, Cultured , Culture Media , Fatty Acids/metabolism , Female , Isomerism , Oleic Acids/analysis , RatsABSTRACT
A study was conducted to determine which fatty acids in plasma membranes of adipose tissue from ruminants are changed when the diet is supplemented with unsaturated fatty acids and to determine the effect of the fat supplement on adipocyte metabolism. Ten sheep were randomly assigned to two isonitrogenous diets containing either no added fat (control) or 5 g soybean oil/100 g diet. Perirenal fat was removed at slaughter, adipocytes isolated by collagenase digestion, and plasma membranes prepared by centrifugation on a Percoll gradient. Feeding soybean oil to the sheep increased (P < 0.05) linoleic acid [18: 2(n-6)] concentration in subcutaneous fat and isolated adipocytes, suggesting partial escape of dietary unsaturated fatty acids from ruminal biohydrogenation. Soybean oil consumption also decreased (P < 0.05) concentrations of myristic acid, arachidonic acid [20: 4(n-6)] and anteiso 17:0 in plasma membranes, but increased (P < 0.05) trans 18:1. Lipogenesis was not affected by diet, but lipolysis tended to be greater (P = 0.07) in sheep fed the soybean oil-containing diet than in those fed the control diet. In ruminants, fatty acids of ruminal origin, namely trans intermediates of biohydrogenation or branched-chain fatty acids of microbial lipid, may account for as much change in the composition of plasma membranes and in cellular metabolism as do the small quantities of unsaturated fatty acids in the diet that escape biohydrogenation.
