Plasma lipoprotein (a) levels in men and women consuming diets enriched in saturated, cis-, or trans-monounsaturated fatty acids

Studies that have shown adverse effects of trans-unsaturated fatty acids on plasma lipoprotein (a) [Lp(a)] levels have used levels of trans-fatty acid that are higher than those in the average U.S. diet. This study was conducted to clarify the effects on Lp(a) of trans-fatty acids levels commonly found in U.S. diets. Lp(a) levels were measured in a double-blind study of 29 men and 29 women who ate 4 controlled diets in random order for 6 weeks each. Fatty acids represented 39% to 40% of energy. The diets were: (1) Oleic (16.7% of energy as oleic acid); (2) Moderate trans (3.8% of energy as trans-monoenes, approximately the trans content of the U.S. diet); (3) High trans (6.6% of energy as trans-monoenes); (4) Saturated (16.2% of energy as lauric plus myristic plus palmitic acids). The Saturated diet lowered Lp(a) levels significantly (by 8% to 11%). Compared to the Oleic diet, the trans diets had no adverse effect on Lp(a) levels when all subjects were considered collectively. A subset with initially high levels of Lp(a) (> or = 30 mg/dL), however, responded to the High trans diet with a slight (5%) increase in Lp(a) levels relative to the Oleic and Moderate trans diets. Thus, in amounts commonly found in the typical U.S. diet, saturated fatty acids consistently decrease Lp(a) concentrations. The adverse effects of replacing cis- with trans-fatty acids are only suggestive and are restricted to high trans intakes in subjects with high Lp(a) levels.     

Divergent incorporation of dietary trans fatty acids in different serum lipid fractions

We isolated from the endogenous polyprenyl-phospho-sugar pool of Mycobacterium smegmatis two mannose-containing compounds, i.e., a partially saturated C35-octahydroheptaprenyl-P-mannose and a fully unsaturated C50-decaprenyl-P-mannose. The relative amount of C35-polyprenyl-P-mannose in mycobacterial cells was comparable to that of decaprenyl- P-pentoses and, at least, an order of magnitude higher than that of C50-decaprenyl-P-mannose. The major form of mycobacterial polyprenyl-P-mannose was structurally characterized by combined gas chromatography-mass spectrometry, fast-atom bombardment tandem mass spectrometry and proton-nuclear magnetic resonance spectroscopy as beta-d-mannopyranosyl-monophospho-(C35)octahydroheptapren ol of which all three isoprene units have Z ( cis ) configuration. The differences in the structure and cellular concentrations of the mycobacterial mannosyl-P-polyprenols reflect distinct biochemical pathways of the two compounds and suggest the existence of specific GDP-Man:polyprenyl-P mannosyltransferases (synthetases) able to distinguish between C35-octahydroheptaprenyl- and C50-decaprenyl- phosphates of mycobacteria. Since the 6'-O-mycoloylated form of C35-octahydroheptaprenyl-P-mannose isolated from M. smegmatis is apparently involved in mycolate rather than mannosyl transfer reactions, we speculate that a catabolic pathway responsible for degradation of C35-P-mannose and recycling C35-octahydroheptaprenyl phosphate might exist in mycobacteria.     

The effect of plasma free fatty acids and long-chain triglycerides on glucose metabolism in uncomplicated falciparum malaria

To investigate the therapeutic potential of increased plasma free fatty acid (FFA) and triglyceride concentrations in hypoglycaemic patients receiving quinine, 32 untreated Thai adults with uncomplicated falciparum malaria were allocated at random to one of 4 regimens: 2 mg/kg/min dextrose infused over 60 min either alone (group A) or with a prior injection of 5000 units of heparin and simultaneous Intralipid infusion (group C), or 4 min/kg/min dextrose alone (group B) or with heparin and Intralipid (group D). Quinine (10 mg/kg) was also infused over 60 min in all cases. In patients of groups A and C, mean changes in plasma glucose concentrations from the beginning to the end of the infusion were 0.1 (SD 0.8) and 1.0 (SD 0.7) mmol/L respectively (P = 0.015). In groups B and D, plasma glucose increased by 1.8 (SD 1.2) and 2.2 (SD 0.4) mmol/L respectively (P < 0.5). Plasma FFA levels fell by approximately 50% during the infusion in groups A and B but increased by a similar percentage in groups C and D. Despite significant mean increases in plasma insulin during the infusion (from 12.2 milliunits (mu)/L in group A to 38.8 mu/L in group D), no rebound hypoglycaemia was observed in any patient during the ensuing 7 h. These data suggest that the glycaemic response to dextrose given at high rates, which match average glucose utilization in a severely ill patient with malaria, is not augmented by increased plasma FFA and long-chain triglycerides. However, this strategy increases the glycaemic efficacy of lower dextrose infusion rates and the combination could, therefore, reduce the volumes of hypertonic dextrose required to prevent hypoglycaemia in severely ill patients in whom optimal fluid balance is crucial.     

Acute effects of dietary fatty acids on the fatty acids of human milk

1. Effects on 5-HT function of sibutramine and its active metabolites, BTS 54 354 and BTS 54 505, were compared with fluoxetine, (+)-fenfluramine and (+)-amphetamine. 2. In vitro sibutramine weakly inhibited [3H]-5-HT uptake into brain synaptosomes. BTS 54 354, BTS 54 505 and fluoxetine were powerful [3H]-5-HT uptake inhibitors, whereas (+)-fenfluramine and (+)-amphetamine were very much weaker. Conversely, whilst sibutramine, its metabolites and fluoxetine did not release [3H]-5-HT from brain slices at < or = 10(-5)M, (+)-fenfluramine and (+)-amphetamine concentration-dependently increased [3H]-5-HT release. 3. Sibutramine and fluoxetine had no effect on 5-hydroxytryptophan (5-HTP) accumulation in either frontal cortex or hypothalamus at doses < 10 mg kg(-1). In contrast, (+)-amphetamine ( > or = 3 mg kg(-1)) reduced 5-HTP in hypothalamus, whilst (+)-fenfluramine (> or =1 mg kg(-1)) decreased 5-HTP in both regions. 4. Sibutramine (10 mg kg(-1) i.p.) and fluoxetine (10 mg kg(-1) i.p.) produced slow, prolonged increases of extracellular 5-HT in the anterior hypothalamus. In contrast, (+)-fenfluramine (3 mg kg(-1) i.p.) and (+)-amphetamine (4 mg kg(-1) i.p.) induced rapid, short-lasting increases in extracellular 5-HT. 5. Only (+)-fenfluramine (10 mg kg(-1)) altered 5-HT2A receptors in rat frontal cortex when given for 14 days, producing a 61% reduction in receptor number and a 18% decrease in radioligand affinity. 6. These results show that sibutramine powerfully enhances central 5-HT function via its secondary and primary amine metabolites; this effect, like that of fluoxetine, is almost certainly mediated through 5-HT uptake inhibition. By contrast, (+)-fenfluramine enhances 5-HT function predominantly by increasing 5-HT release. (+)-Amphetamine, though weaker than (+)-fenfluramine, also enhances 5-HT function by release.     

Plasma and erythrocyte alpha-tocopherol and plasma retinol concentrations in term infants fed formula enriched with long-chain polyunsaturated fatty acids

OBJECTIVE: To assess the effect of long-chain polyunsaturated fatty acids (LCPUFA)- and vitamin E-supplemented formula feeding on erythrocyte and plasma alpha-tocopherol (VE), and plasma retinol (VA) concentrations in neonates and to compare these values with those found in infants feeding on infant formula without LCPUFA or breast milk SETTING: University Hospital of Granada, Spain. SUBJECTS: 49 full-term infants. DESIGN AND INTERVENTION: Subjects who chose not to breast feed were fed either (i) unsupplemented infant formula (F) or (ii) infant formula supplemented with LCPUFA and vitamin E (FL). Alpha-tocopherol and retinol were measured at 7 days, 1 month and 3 months. RESULTS: Plasma and erythrocyte VE concentrations and plasma VE/total lipids ratio increased significantly in all groups at 1 month of life (P < 0.05), but did not change significantly between 1 month and 3 months in any group (P > 0.05). Erythrocyte VE and VA retinol concentrations were higher in infants fed an infant formula than in breast milk-fed infants at 1 month of life (P < 0.05). Finally, there were no significant differences in plasma or erythrocyte VE levels, plasma VA or plasma VE/total lipid ratio between any groups at 3 months of life (P > 0.05). CONCLUSION: Infants fed on LCPUFA- and vitamin E-supplemented infant formula for 3 months have similar vitamin E and A status to infants fed on breast milk or infant formula without LCPUFA supplementation.     

Regulation of tumour cell fatty acid oxidation by n-6 polyunsaturated fatty acids

The aim of this study was to evaluate acute effects of ethyl tert-butyl ether (ETBE) in man after short-term exposure. ETBE may in the future replace methyl tert-butyl ether, a widely used oxygenate in unleaded gasoline. Eight healthy male volunteers were exposed to ETBE vapor for 2 h at four levels (0, 5, 25, and 50 ppm) during light physical exercise. The subjects rated irritative symptoms, discomfort, and central nervous system effects in a questionnaire. Ocular (eye redness, tear film break-up time, conjunctival epithelial damage, and blinking frequency), nasal (acoustic rhinometry and analysis of inflammatory markers and cells in nasal lavage fluid), and pulmonary (peak expiratory flow, forced expiratory volume in 1 s, forced vital capacity, vital capacity, and transfer factor) measurements were performed. Significantly increased ratings of solvent smell (p = 0.001, repeated-measures ANOVA) were seen during exposures and correlated to exposure levels. Furthermore, significantly elevated ratings of discomfort in throat and airways were seen during and after 50 ppm compared to the control exposure (p = 0.02). Increased nasal swelling (p = 0.001) and blinking frequency (p = 0.01) were noted at all exposure levels, but their magnitudes were not related to exposure levels. A slightly impaired pulmonary function was seen at 25 and 50 ppm, since forced vital capacity (p = 0.02) and vital capacity (p = 0.04) differed significantly from the clean air exposure. Although the impairments seemed to fall within normal inter- and intraindividual variation and have no clinical relevance as such, it cannot be excluded that other individuals may react more severely than eight healthy male volunteers in this study.     

Modulation of exercise-induced immunosuppression by dietary polyunsaturated fatty acids in mice

The possible interaction between intense exercise, known to suppress the immune response, and nutritive factors, such as polyunsaturated fatty acids (PUFA), was examined in inbred female C57Bl/6 mice. The animals received for 8 wk either a natural ingredient diet or a diet supplemented with 10 g/100 g linseed oil containing over 50% of 18:3 (n-3) alpha-linoleic acid. Other groups received PUFA containing only traces of 18:3 (n-3) fatty acid; beef tallow, containing mostly 18:1 (n-9) saturated fat, safflower oil, an 18:2 (n-6) PUFA, and fish oil, containing longer chain (n-3) PUFA. Each dietary group was divided into two subgroups: sedentary diet controls and exercised animals. Exercise consisted of continuous swimming at high intensity until exhaustion. It was shown in three separate experiments that (1) the primary humoral response to sheep red blood cells, determined by the plaque-forming cell (PFC) assay, was affected by PUFA diet in sedentary animals in the order beef tallow > control diet > safflower oil > fish oil > linseed oil, and (2) the PFC response was suppressed by the exhaustive exercise, as compared to sedentary controls, except for animals fed 18:3 (n-3) linseed oil, where the normal response was noted. Phagocytosis of fluorescent microspheres by peritoneal macrophages, determined by flow cytometry, was significantly lower in exercised animals receiving the linseed oil diet, whereas other diets either increased or did not significantly change the macrophage phagocytic activity, compared to the sedentary diet controls. Spleen lymphocyte subsets were unchanged in exercised animals except for a marked shift from the lymphoid peak toward the erythroid peak. Generally, our data showed a marked immunomodulatory effect of 18-3 (n-3) alpha-linoleic acid on the exhaustive exercise-related immunosuppression, as compared to the effects of other selected PUFA.     

Distribution of plasma phosphatidylcholine molecular species in rabbits fed fish oil is modulated by dietary n-6 fatty acids

Natural occurrence of Fusarium mycotoxins (trichothecenes and fumonisins) and aflatoxin B1 (AFB1) were surveyed in 32 corn samples, harvested in 1993 and randomly sampled in 1994 in several districts of Hanoi, Vietnam. Corn samples were first milled into fine powder, extracted with methanol-water (3:1) and the crude extracts obtained from the same samples were used for the simultaneous analysis of the trichothecenes such as nivalenol (NIV), deoxynivalenol (DON), and T-2 toxin (T-2) by gas chromatography/mass spectrometry (GC/NS); fumonisins B1 (FB1), B2 (FB2), and B3 (FB3) by high-performance liquid chromatography (HPLC) with a flourescence detector; and AFB1 by and ELISA kit based on a monoclonal antibody. The data revealed that 14, 8, 4, 3, and 2 out of 15 corn kernel samples were positive for AFB1, FB1, FB2, FB3, and NIV with the average levels being 28, 1, 101, 276, 232, and 858 ppb, respectively, and neither DON nor T-2 were detected. As for the other 17 samples of corn powder, 13, 15, 12, 10, 4 and 2 were positive for AFB1, FB1, FB2, FB3, DON, and NIV with the average being 30, 780, 289, 176, 3, 170, and 1,365 ppb, respectively, and T-2 was not detected. Although their positive rates and levels fell in the ranges reported elsewhere, it was found for the first time that the Fusarium toxins (NIV, DON, and fumonisins) and an Aspergillus toxin (AFB1) were naturally co-contaminated in selected samples of corn produced in north Vietnam.     

Depression of lipogenesis in swine adipose tissue by specific dietary fatty acids

The objective of this study was to document the influence of specific dietary fatty acids on rates of lipid synthesis and sensitivity to insulin in porcine adipose tissue. Weanling pigs were assigned to one of six groups, and each group was fed diets containing 10 g/100 g of added cornstarch or 10 g/100 g of added fatty acid. The fatty acid-enriched diets contained either a combination of 14:1 plus 16:1 (14:1/16:1 diet), 16:0, 18:0, 18:1, or 18:2 (n-6). With the exception of the cornstarch diet, all diets contained approximately 35% 14:0. Subcutaneous adipose tissue samples were collected at slaughter from the area overlying the first cranial vertebra. Fresh samples were incubated for 2 h in 20 mM glucose and 0, 10, 100 or 1,000 microU/mL of porcine insulin. The smallest adipocytes were observed in adipose tissue from pigs fed the 16:0 or 18:2 diets. Glucose incorporation into lipids was greater (P < .05) in adipose tissue from cornstarch-fed pigs than in adipose tissue from the other treatment groups. Lipogenesis was 67, 53, 35, 32, and 20% lower (P < .05) in adipose tissue from 16:0-, 14:1/16:1-, 18:0-, 18:2-, and 18:1-fed pigs, respectively, than in adipose tissue from the cornstarch-fed pigs. Insulin increased lipogenesis by 19% (P < .05) in adipose tissue from the cornstarch-fed pigs and by 15 to 40% (P < .05) in adipose tissue from the 14:1/16:1-fed pigs. Insulin did not stimulate lipogenesis (P > .4) in adipose tissue from pigs fed the 16:0, 18:0, or 18:1 diets. The data suggest that fatty acid chain length and unsaturation are determinants in the effects of dietary fat and insulin on de novo lipogenesis.     

Regulation of pancreatic and gallbladder functions by intraluminal fatty acids and bile acids in man

The effects of intraduodenal glycerol, fatty acid (FA) chain length and FA loads, and bile acid (BA) concentrations on pancreatic and gallbladder function were investigated in 31 healthy volunteers by a perfusion method. FA absorption rates in the duodenum and proximal jejunum were measured simultaneously. Pancreatic and gallbladder responses were augmented by increasing FA chain length and FA loads until the "maximal" secretory capacity of the pancreas and gallbladder emptying was attained. Glycerol had no effect. Raising BA concentrations above the critical micellar concentration accelerated FA absorption rates but decreased the magnitude of pancreatic and gallbladder responses to FA. Higher BA concentrations exerted an opposite effect, slowing FA absorption and increasing pancreatic and gallbladder responses. Indeed, a significant, inverse correlation was found between FA absorption and pancreatic and gallbladder responses to FA, suggesting a relationship between the length of intestine exposed to FA and the amount of cholecystokinin (and/or other neurohormonal factors) released, which stimulates pancreatic secretion and gallbladder contraction.     

Utilization of supplemental triglycerides or free fatty acids by calves from 4 to 10 weeks of age

Our objectives were 1) to determine the effects of increased availability of 2-monoglycerides from triglyceride digestion on absorption of free fatty acids (FFA) and utilization of nutrients by calves of weaning age and 2) to determine whether absorption of FFA increased with age. Thirty-two male Holstein calves were fed milk replacer until d 28 of age and then were assigned, to one of four treatments: 1) control, 80 g/d of whey protein concentrate in 2 L of water; 2) control plus 120 g/d of lard triglycerides; 3) control plus 90 g/d of lard triglycerides plus 30 g/d of lard FFA (low FFA); and 4) control plus 60 g/d of triglycerides plus 60 g/d of lard FFA (high FFA). Treatments were fed by nipple pail twice daily to maintain closure of the esophageal groove. Intake of calf starter (available continuously) was decreased by supplemental fats and tended to be lower for calves fed high FFA than for those fed low FFA. Digestibilities of total C18 fatty acids and total fatty acids were lower for calves fed triglycerides than for calves fed FFA, but digestibilities were not affected by age. Energy digestibility was greater for fat treatments than for controls and was greater for FFA treatments than for triglycerides, but intakes of digestible energy and metabolizable energy did not differ. Absorbed N was greater for calves on the control treatment than for those on the fat treatments, but retained N and average daily gain did not differ. Digestibility of fatty acids was greater for calves fed mixtures of triglycerides and FFA than for those fed triglycerides alone, but digestibility did not change during wk 6 to 10 of age.     

Effect of dietary trans fatty acids on cholinesterase and monoamine oxidase activities in different organs of rats

The effect of trans fatty acids and essential fatty acid deficiency upon the activities of cholinesterase and monoamine oxidase in livers, hearts, brains and lungs of rats was studied. This study was performed using three groups of male albino rats. Two out of these three groups were fed essential fatty acid low diets containing 10% hydrogenated coconut oil (HCNO) or margarine stock (MS, partially hydrogenated soybean oil) while the third group was fed an adequate supply of essential fatty acids through a diet containing 10% corn oil (CO). In the group of rats fed HCNO the activities of cholinesterase and monoamine oxidase in their livers, hearts, brains and lungs were not significantly different from those of the control group fed CO. In the group of rats fed MS, the activity of cholinesterase was significantly decreased in the livers, hearts and brains, but not affected in the lungs, while the activity of monoamine oxidase was significantly decreased in the livers and hearts but not affected in the brains and lungs as compared to the control group fed CO. The levels of serum total lipids, total cholesterol and triglycerides were elevated in both groups of rats fed HCNO or MS than in the group fed CO, but this elevation was more highly significant in the group fed MS than in the group fed HCNO.     

Removal of low-density lipoprotein from plasma by adsorption increases bradykinin and plasma nitric oxide levels in patients with peripheral atherosclerosis

Low-density lipoprotein (LDL) adsorption using a dextran sulfate cellulose column is brought about by electrostatic binding between the positive charges of apolipoprotein B in LDL and the negative charges of dextran sulfate cellulose. There is general agreement that the initial contact phase in the coagulation pathway may be activated by a negatively charged surface such as dextran sulfate cellulose, resulting in the generation of bradykinin. We investigated whether the increase in the generation of bradykinin during LDL adsorption is accompanied by the activation of endogenous production of nitric oxide (NO) in patients with peripheral atherosclerosis. LDL adsorption therapy was repeated ten times over a period of 3 months in ten peripheral atherosclerosis patients. Treatment significantly reduced serum total cholesterol and LDL cholesterol levels. This was associated with a significant improvement in Fontaine's classification and ankle pressure index. We also measured the kinin-kallikrein system and plasma levels of NO in the same patients. The results showed that coagulation factors of the intrinsic pathway including high-molecular-weight kininogen and prekallikrein decreased markedly after initial adsorption compared with the levels before treatment. There was a marked increase in bradykinin and NO concentrations after the initial adsorption, compared with their levels before adsorption. Our results suggest that the generation of bradykinin and increased plasma levels of NO may contribute to the improvement in peripheral circulation after LDL adsorption in peripheral atherosclerosis patients.     

Digestion and absorption of dietary triglycerides

This review is an attempt to put in order some facts obtained during twenty-five years of studying the digestion and absorption of dietary triglycerides. After a short history of this problem to show the progress of research, the active mechanism of pancreatic lipase on triglycerides is explained: this enzyme specifically hydrolyses fatty acids esterified with the primary hydroxyl groups of glycerol, forming 2-monoglycerides and free fatty acids. This fact is very important for the further process involving formation and composition of intraluminal micelles, along with absorption and de novo synthesis of triglycerides inside enterocytes. Several different points are disscussed: - Short or medium chain-length fatty acids are hydrolyzed more easily than long-chain ones; after their absorption, the former go to the liver by the portal circulation and the latter go to lymph. - An evaluation of the extent of digestive hydrolysis has been given by means of doubly-labelled triglycerides (glycerol and fatty acids), and by comparing isotope ratio values of lymph and dietary triglycerides. - The physico-chemical state that intraluminal fats are in is discussed; 2-monoglycerides and fatty acids released from triglyceride hydrolysis form macromolecular agregates with biliary salts and phospholipids, cholesterol, and dietary phospholipids (or their degradation products). These are termed mixed micelles and are absorbed by diffusion. - Many recent morphological studies carried out with the electron microscope are indicated. - Two metabolic pathways for triglyceride resynthesis in the mucosal cells have been established, one from free fatty acids and alpha-glycerophosphate, the other from 1 or 2-monoglycerides. Phosphatidic acids are involved in this resynthesis, but the role of other phospholipids is almost certainly not so narrowly limited. - The apparent digestibility of fats according to their fatty acid composition, and mainly according to the location of the fatty acids in the initial triglyceride molecule, is discussed. - The nature and importance of endogenous fatty acids in the digestive tract and lymph is discussed. - The lipid and fatty acid composition of lymph chylomicrons is given. - The importance of lymph lipid particles (average diameter 0,05 less than or equal mum) for transport of exogenous fatty acids is indicated. The biochemical mechanism involved in the digestion and absorption of dietary lipids are now well known, but some processes are still uncertain, particularly the physico-chemical state of intraluminal lipids, the role of enterocyte membranes, the synthesis and secretion of digestive enzymes, and the role of intestinal flora.     

Enrichment of hen eggs with n-3 long-chain fatty acids and evaluation of enriched eggs in humans

Eggs enriched with n-3 polyunsaturated fatty acids (PUFAs) were produced by hens fed diets containing fish oil or a combination of fish and vegetable oils. In a sensory evaluation, 78 untrained volunteers could not distinguish between ordinary and enriched eggs. Storage life was also not significantly different between egg types. A food intake survey of 4 groups of 14 subjects each who consumed 7 eggs/wk for 24 wk showed that intakes of the major dietary components were not significantly different for 4 different egg types. Mean (n = 56) plasma cholesterol and triacylglycerol concentrations were not significantly different at the start and finish of the study. Body weight and HDL concentrations increased during the study (P < 0.05). For the last 2 wk of the experiment (weeks 23-24), mean egg consumption was increased from a total of 14 to a total of 21 eggs, resulting in a small increase in plasma triacylglycerols only. There were no significant differences (P > 0.05) in body weight, blood pressure, or plasma lipid components among treatment groups consuming the 4 different egg types. Blood samples taken after 16 and 22 wk from fasted subjects showed significant increases in eicosapentaenoic acid, docosahexaenoic acid, and total n-3 PUFAs in subjects consuming enriched eggs compared with controls. In addition, the ratio of n-6 to n-3 PUFAs in plasma was significantly reduced from 12.2:1 to 6.5-7.7:1 in subjects consuming enriched eggs compared with controls. Consumption of only one enriched egg daily can contribute substantially to the recommended daily intake of n-3 PUFAs.     

Enzymatic hydrolysis of anchovy oil: production of glycerides enriched in polyunsaturated fatty acids

In an attempt to produce the polyunsaturated fatty acid (PUFA)-enriched glycerides, commercially available Turkish anchovy oil (PUFA content of 27%), was hydrolyzed with 1,3-specific Rhizomucor miehei lipase. After the hydrolysis, the triglyceride (TG), diglyceride (DG), monoglyceride (MG), and free fatty acid (FFA) composition of the reaction mixture was determined, and fatty acid components of these fractions were analyzed. R. miehei lipase released PUFA extremely slowly, resulting in their accumulation in the TG and DG fractions, especially in TG. The PUFA content in the glyceride mixture (including TG, DG, and MG) increased as hydrolysis progressed. The effects of operational parameters (pH, temperature, time, and enzyme concentration) on the extent of hydrolysis were investigated. Based on these results, optimal reaction conditions were established. At optimal conditions (pH 4.0, 35 degrees C, 3 h, and enzyme concentration of 500 U/g oil), the level of PUFA in the glyceride mixture was raised to 40%. The individual TG and DG fractions contained 45 and 30% PUFA, respectively. Less than 2% of the total PUFA was lost in the FFA fraction.