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Simultaneous determination of fat-soluble vitamins and provitamins in dairy products by liquid chromatography with a narrow-bore column.

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of vitamins A, D2, D3, E and K1, retinyl acetate, retinyl palmitate, tocopherol acetate, ergosterol and 7-dehydrocholesterol in milk and butter. Narrow-bore columns are recommended because this alternative provides a good separation and efficiency, plus greater economy and sensitivity. Detection limits for individual vitamins range from 0.14 to 6.9 ng. All vitamins are separated in less than 33 min. For the simultaneous determination of these vitamins and provitamins we use two sample pre-treatment methods, a liquid-liquid extraction with hexane or a solid-phase extraction with a C18 cartridge. Recovery studies show good results for all solutes (84-108% and 85-108% for milk and butter, respectively) and the intra-day coefficients of variations range from 1.6 to 4.5%. These methods permit the simple determination of fat-soluble vitamins using a small sample volume.

Animals↗

The alpha and gamma tocopherol levels in serum are influenced by the dietary fat quality.

BACKGROUND: Alpha tocopherol in serum is thought to be of importance in protecting lipids against oxidation and low serum levels of alpha tocopherol has been suggested to increase the risk for coronary heart disease. However, low levels of gamma, rather than alpha, tocopherol have been found in patients with manifest coronary heart disease and in populations with a high incidence of coronary heart disease. AIM: The aim of this study was to determine the tocopherol concentrations in serum after two diets with identical nutrient content but with different fat quality, enriched in butter and rapeseed oil-based fats, respectively. METHOD: Twenty moderately hyperlipidemic, healthy subjects (six females and 14 males) participated in this double-blind cross-over study, where two isoenergetic diets were given in a randomized order during two 3-week periods, interrupted by a wash-out period of 3-4 weeks. RESULTS: The lipid-corrected serum concentrations of alpha and gamma tocopherol increased during the diet rich in rapeseed oil (by 7 and 23%, respectively) compared with on the baseline diet (P < 0.001), while these concentrations decreased (by 5 and 37%, respectively, P < 0.01) during the diet rich in saturated fat. The ratio between alpha and gamma tocopherol decreased significantly during the rapeseed oil diet (-23%, P < 0.01) and increased (+46%, P < 0.001) during the butter diet. CONCLUSION: Alpha and gamma tocopherol levels in serum are influenced by the type of fat used in the diet. The most unexpected finding is that the lipid-adjusted gamma tocopherol concentration significantly decreased by 37% during a diet rich in saturated fat with an increased ratio between alpha and gamma tocopherol, similar to the situation found in CHD patients.

Aged↗

Response of extrapancreatic immunoreactive glucagon to intraluminal nutrients in pancreatectomized dogs.

To investigate the response of extrapancreatic glucagon to intraluminal stimuli, nutrients were administered to normal and pancreatectomized dogs through a stomach tube in a fully conscious state after an overnight fast. Plasma immunoreactive glucagon was determined with antisera specific and nonspecific to glucagon and was designated as IRG and total IRG, respectively. Oral glucose load elicited a decrease in plasma IRG and a remarkable rise of plasma total IRG in a group of 6 pancreatectomized dogs, as in the control dogs. When arginine was given, both plasma IRG and total IRG significantly increased in a group of 5 pancreatectomized dogs, while only total IRG rose significantly in the normal control dogs. Butter load did not reveal any changes in plasma IRG and total IRG in a group of 5 pancreatectomized dogs, whereas only total IRG increased in the normal control dogs. It is concluded that extrapancreatic glucagon responds to intraluminal administration of nutrients, as pancreatic glucagon does. In addition, gut glucagon-like immunoreactivity increased following glucose or arginine ingestion in pancreatectomized dogs. Furthermore, the failure in response of plasma IRG and total IRG to butter load in pancreatectomized animals suggests that its intraluminal hydrolysis is important in the secretion of extrapancreatic immunoreactive glucagon.

Animals↗

Pesticide residues in the Canadian Market Basket Survey--1992 to 1996.

Market basket food samples from six Canadian cities collected from 1992 to 1996 were analysed for pesticide residues. One hundred and thirty-six composites were prepared for each city, representing 99% of the Canadian diet. Residues were found most frequently in peanut butter and butter. DDE, malathion and captan occurred most frequently, while the fungicides chlorothalonil, dicloran and captan were present in the highest concentrations. Processed commodities contained fewer residues and at lower concentrations than the raw products. No residues were detected in either milk or soy-based infant formula. Of the infant foods sampled, fruit contained both the greatest number and highest concentrations of pesticides.

Arachis↗

Levels of specific polychlorinated biphenyl congeners in fatty foods from five Canadian cities between 1986 and 1988.

A total of 155 fatty food composites from five major Canadian cities were analysed for 36 selected polychlorinated biphenyl (PCB) congeners. Total PCB congener levels of more than 1 ng/g (wet weight) were found in fresh water fish, canned fish, butter, marine fish and cheese, with fresh water fish (18.7 ng/g) containing 5 to 10 times more total PCBs than the other four food commodities. Milk (2%), cooking oils/salads and canned meat soup contained less than 0.1 ng/g (wet weight). The observed total PCB residue levels in fatty foods were well within the Canadian guidelines for fish, dairy products, poultry, eggs and beef. The most predominant congeners found in fish, butter, cheese, meat and poultry were the 2,2',4,4',5-, 2,3',4,4', 5-pentachloro-, 2,2',3,4,4',5'-, 2,2',4,4',5,5'-hexachloro- and 2,2',3,4,4',5,5'-heptachlorobiphenyls. The predominant congener pattern was not always evident in other dairy products (e.g. ice cream) and processed foods (e.g. canned meat soup). An attempt was made to relate the intake of PCB congeners from fatty foods with congener levels found in adipose tissue of Canadians. The estimated daily intake of some specific PCB congeners from fatty foods, in most cases accounted for more than half the total deposit of these congeners in adipose tissue of Canadians.

Adipose Tissue↗

Polychlorinated biphenyl residues in various fatty foods consumed in Guipúzcoa, the Basque Country (Spain).

Three congeners of polychlorinated biphenyls (PCBs)--the 2, 2', 3, 4, 4', 5'-, 2, 2', 4, 4', 5, 5'-hexachloro, and 2, 2', 3, 4, 4', 5, 5'-heptachlorobiphenyls--were determined in 18 samples of butter, 44 of fish (hake and anchovy) and 24 of egg, all of which came from markets in Guipúzcoa. The total content as Araclor 1260 was also determined. Quantitation was carried out by gas chromatography in a capillary column and electron capture detector. The total content in butter ranged from 20 to 60 ng/g (fat basis), in fish from 10 to 250 ng/g (edible part), and in eggs from 1 to 10 ng/g (edible part). None of these exceeded the tolerance limits established by the Canadian Government Guideline and USFDA.

Animals↗

Regional differences in consumption of 103 fat products in Belgium: a supermarket-chain sales approach.

OBJECTIVE: In Belgium, important regional differences in mortality exist which have been linked to differences in dietary consumption patterns. METHODS: To study regional food consumption in Belgium, sales data of 103 spreading and cooking fat products in 110 branch stores of a major supermarket chain (Colruyt) for 12 months (1991-92) were analyzed. RESULTS: Sale of more ordinary and polyunsaturated spreading margarine and of more polyunsaturated low-fat spread in the north in combination with a greater sale of butter and dairy low-fat spread in the south resulted in a P/S-ratio of 0.99 in the north vs. 0.40 in the south (p < 0.001) and a U/S-ratio of 1.93 vs. 1.10 (p < 0.001) for spreading fats. The P/S- and U/S-ratios of cooking fats were lower in the north (ns). Mortality data correlated positively with the sale of butter and dairy low-fat spread and negatively with sales data of spreading margarine, polyunsaturated spreading margarine, and polyunsaturated low-fat spread (all p < 0.001). CONCLUSION: Sales data from supermarket chains provide useful information on regional fat consumption and offer interesting perspectives of trends over time.

Belgium↗

Effect of high crude fiber intake on transit time and the absorption of nutrients in South African Negro schoolchildren.

Transit time (using carmine as marker) averaged about 9.5 hours in rural South African Negro children aged 9-12 years, when ingesting a mean of 10 g crude fiber per diem on their everyday diet. A: in subgroups of 40-50 pupils, supplements of maize and wheat bran, and of wholemeal bread--each affording about 2 g fiber--also of oranges, affording 4 g fiber, decreased transit time only slightly, to about 8.5 hours. Hence, pupils current daily fiber intake, in its effect on transit time, is already acting maximally. B: mean respective fecal nitrogen and fat concentrations on everyday diet were not significantly affected when the subgroups ate one of the following fibre-free foods: 1) 40 g protein from skim milk; 2) 40 g fat from butter; 3) 40 g carbohydrate afforded by sugar. Additionally, on their usual diet, also following the ingestion of 40 g uncooked maize flour, starch was extremely seldom detected in feces. C. feces collections for 5-day periods were made on small groups of pupils (8 per group) when consuming first their everyday diet, and next when including each day one of the following: 1) 40 g protein from skim milk; 2) 40 g fat from butter; 3) 40 g sugar. On these regimens, there were no significant changes, compared with respective data on everyday diet, in mean amount of dry feces excreted daily, in concentrations of nitrogen and fat, nor in amounts of nitrogen and fat excreted daily. Yet a daily supplement of 350 g oranges (5-7 oranges), although virtually nitrogen- and fat-free but containing 4 g fiber, resulted in slightly shorter transit time but highly significant increases in mean amounts of dry feces, nitrogen and fat excreted daily. Observations suggest that the protein, fat and carbohydrate (excluding unassimilable fiber) in the supplements were virtually fully digested and absorbed. Discussion suggests that this also applied to nutrients in the everyday diet. It is judged that the large stools voided by these people (and others consuming diets high in fiber) are composed, apart from unassimilable fiber, almost wholly of endogenous products.

Animals↗

Glycemic responses to foods: possible differences between insulin-dependent and noninsulin-dependent diabetics.

The effect on the blood glucose response of varying the amount (25 or 50 g) and type (bread or beans) of carbohydrate (CH2O) in test meals and of adding fat and protein was examined in a group of insulin dependent and noninsulin dependent diabetic volunteers. With noninsulin-dependent diabetics, the blood glucose area after a half bread portion was 48% that of the full bread meal (p less than 0.001). White pea beans (50 g CH2O) gave a blood glucose response of 41 +/- 5% (p less than 0.001) that of bread (50 g CH2O). A combined meal of bread (25 g CH2O) and beans (25 g CH2O) gave a blood glucose response of 60 +/- 8% of the full bread meal (p less than 0.005) and similar to that expected. Addition to bread of butter and skim milk cheese either singly or in combination had little effect on the glycemic response although a reduced rise was seen after addition of fat and protein as peanut butter. Insulin-dependent diabetics tended to respond similarly to noninsulin-dependent diabetics but their greater variability indicates that other factors in addition to the nature of the food may also be major determinants of their glycemic responses to foods.

Adult↗

Divergent lipoprotein responses to fish oils with various ratios of eicosapentaenoic acid and docosahexaenoic acid.

To determine if the ratio of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids in fish oil had an effect on plasma lipid responses, we randomly fed eight normolipidemic men three 36%-fat diets containing primarily butter, EPA-rich pollock oil, or DHA-rich tuna or salmon-blend oils. Plasma EPA and DHA reflected the amounts in the diets. Compared with values for the butter diet, very-low-density lipoprotein (VLDL) triglycerides decreased equally (71-78%) with all diets; low-density lipoprotein (LDL) cholesterol (LDL-C) and apolipoprotein B decreased 26% and 13%, respectively, on the tuna and salmon-blend oil but did not change (-1%) and increased 19% with the pollock diet; high-density lipoprotein cholesterol (HDL-C) and lipoproteins A-I and A-II decreased with all diets but more with the pollock diet than with the tuna and salmon diets. The 23-31% decrease in total cholesterol on the tuna and salmon diets resulted mostly from decreased LDL-C whereas the 16% decrease on pollock oil resulted mostly from a decrease in HDL-C.

Butter↗

Plasma clearance of chylomicrons from butterfat is not dependent on saturation: studies with butterfat fractions and other fats containing triacylglycerols with low or high melting points.

BACKGROUND: Dietary fats influence plasma lipids, and changes in the clearance and metabolism of postprandial lipoproteins can affect atherosclerosis. Butterfat is considered hypercholesterolemic but contains a multitude of constituent fatty acids. OBJECTIVES: We determined triacylglycerol and cholesteryl ester clearances of lymph chylomicrons derived from butterfat, fractions of butterfat, and other dietary fats. METHODS: Radiolabeled lymph chylomicrons resulting from the intestinal absorption of different fats were reinjected into recipient rats to measure plasma clearance. Plasma clearance of [14C]triacylglycerol was used as an indicator of chylomicron lipolysis whereas clearance of [3H]cholesteryl ester was used as an indicator of chylomicron remnant removal. RESULTS: [3H]Cholesteryl ester clearance was slower from chylomicrons derived from a solid, high-saturated-butterfat fraction than from whole butterfat, but clearance of chylomicrons from other fractions did not correlate with the fractions' saturated fatty acid contents. Clearance of cholesteryl esters in chylomicrons derived from cocoa butter, palm oil, and butterfat was slower than clearance of cholesteryl esters in chylomicrons derived from safflower oil. Hepatic uptakes of cholesteryl esters were generally lower for chylomicrons from all butterfat fractions, cocoa butter, and palm oil. CONCLUSIONS: In contrast with minor effects on the lipolysis of chylomicron triacylglycerols, chylomicron remnant removal was strongly influenced by the type of dietary fat, with slower cholesteryl ester clearances for saturated fats with higher melting points. However, remnant removal and hepatic uptake of chylomicrons from whole butterfat and fractions of butterfat were not correlated with fat saturation. The mechanisms of this apparent paradox remain unknown but may be attributable to acyl arrangements in the lipid classes of chylomicrons that influence the association with apolipoproteins and receptors and hence remnant removal.

Analysis of Variance↗

Effect of type of dietary fat, cholesterol and chenodeoxycholic acid on gallstone formation, bile acid kinetics and plasma lipids in squirrel monkeys.

To explore the effect of type of dietary fat, cholesterol and chenodeoxycholic acid on gallstone formation, bile formation, bile composition, bile acid kinetics and plasma lipids in squirrel monkeys, 39 monkeys were studied using seven different diets. Safflower oil, a highly unsaturated fat, added to a diet with cholesterol resulted in at least as high an incidence of cholesterol gallstones as butter added to the same diet. On the other hand, diets with high levels of saturated or unsaturated fat without cholesterol did not result in gallstone formation. Dietary chenodeoxycholic acid (0.1%) did not reduce the incidence of cholesterol gallstones, although the proportion of bile acids as chenodeoxycholic acid increased. Gallbladder biles from monkeys fed semipurified diets with cholesterol had a significantly higher lithogenic index than the comparable groups without cholesterol. Pool sizes of bile acids in all semipurified diet groups were reduced and the lithogenic indices were increased compared with the group fed a commercial feed. Dietary chenodeoxycholic acid caused a decrease in plasma cholesterol in butter groups and an increase in triglyceride concentrations in safflower groups. Diet infuences bile composition and bile acid kinetics, as well as the incidence of gallstones, in squirrel monkeys.

Animals↗

Dietary (n-3) fat and cholesterol alter tissue antioxidant enzymes and susceptibility to oxidation in SHR and WKY rats.

Previously, 8% fish oil blend diets, compared to butter and soybean oil blend diets, reduced specific antioxidant enzyme activities and tissue susceptibility to in vitro oxidative stress in spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats. Moreover, high cholesterol (5.0 g/kg diet) diets protected against in vitro tissue lipid oxidation. In this study, we hypothesized that 160 g fat/kg diet as blends of (n-6) or (n-3) oils and cholesterol would alter antioxidant enzyme activities and thus increase tissue susceptibility to oxidation. The effects of diet blends of saturated (butter, B), (n-6) (soybean oil, SBO) or (n-3) (menhaden oil, MO) oils with cholesterol (0.5 or 5.0 g/kg) on systolic blood pressure (SBP), plasma lipids, antioxidant enzymes and susceptibility to oxidation were examined in SHR and WKY rats. SBP at 13 wk of age was greater (P < 0.001) in SHR than in WKY rats, but was not affected by diets. Plasma cholesterol and triacylglycerols were decreased (P < 0.001) by MO diets. Hepatic glutathione reductase activities were reduced (P < 0.001) in SBO-fed SHR and enhanced in SBO- and MO-fed WKY rats. Glutathione levels were reduced (P < 0.001) in RBC and enhanced (P < 0.001) in livers of MO-fed rats. Lipid oxidation was enhanced (P < 0.001) in red blood cells (RBC) from SBO groups, and hearts and livers of MO groups. High cholesterol diets reduced (P < or = 0.001) susceptibility to lipid peroxidation in RBC and liver of SHR and WKY rats. Greater amounts of dietary (n-3) fat enhance tissue susceptibility to oxidation, which can be modulated by increased dietary cholesterol in SHR and WKY rats.

Animals↗

Kinetics of viral replication in experimental rotavirus infection: effects of high dietary fat.

Epidemiological studies suggest that dietary fat can affect the frequency and severity of diarrhoeal illnesses in childhood. To study the effects of dietary fat on the kinetics of rotavirus antigen, 3-week-old mice of rotavirus-seronegative mothers were fed isocaloric diets with 40% of the total calories as fat [either butter (high saturated fat), olive oil (high monounsaturated fat), or corn oil (high polyunsaturated fat)] with one group on low fat (10% of calories) standard diet as controls. Seven-day-old mice from the first litter were killed and their stomach contents (milk) analyzed for total fatty acids. Seven-day-old mice from the second litter were inoculated with EDIM rotavirus, and some were killed at 48 h and others at 120 h postinoculation. The stool was removed from the distal colon of each mouse and examined for evidence of diarrhoea. The small intestine was removed, the contents washed, and the intestine divided into equal thirds. All stool and tissue samples were made to 10% (wt/vol), homogenised, and tested for rotavirus antigen by ELISA. We found that (a) dietary fat markedly altered the fatty acid profile of milk, (b) high saturated dietary fat (butter) delayed the onset of diarrhoea, reduced the excretion of rotavirus antigen in stool, and reduced mucosal antigen in the lower third of the small intestine at 120 h compared to 48 h postinoculation, (c) high polyunsaturated dietary fat initially increased the frequency of diarrhoea and viral antigen in the middle and lower segment at 120 h, followed by a reduction in rotavirus excretion in stool.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

A complex biochemical modulation of intestinal ion transport in rats fed on high-fat diets.

This study investigates the relationship between dietary fat and ion transport in rat intestine. Animals were fed isocaloric diets with high fat content as corn oil, evening primrose oil (Efamol), fish oil, Efamol/fish oil, olive oil, coconut oil, and butter. A low-fat (corn oil) diet was used in a control group. Biopsies of the small intestine from these animals were mounted in an Ussing chamber, and the intestinal mucosal to serosal ion transport was measured as short-circuit current (SCC). The SCC was stimulated in rats fed on diets high in polyunsaturated fatty acids such as corn oil, Efamol, fish oil, and Efamol/fish oil. SCC was also stimulated in rats fed on diets high in monosaturated fatty acids such as olive oil. Animals fed on diets high in saturated fatty acids such as coconut oil and butter, on the other hand, showed an inhibition in the SCC. To determine whether the effect of membrane lipids was mediated by a differential effect on membrane receptor proteins, the response to secretogogue challenge was studied. Biochemical agents and secretogogues including acetylcholine, noradrenaline, dibutyryl-cAMP, calcium ionophore A2318, and prostaglandin E2 were analysed and compared. These agents were able to influence the SCC, showing responses with marked differences between diet groups.

Animals↗

Serum lipid effects of a monounsaturated (palmitoleic) fatty acid-rich diet based on macadamia nuts in healthy, young Japanese women.

1. Recent studies have identified potential beneficial effects of eating nuts, most of which have substantial amounts of monounsaturated fatty acids (MUFA). Macadamia nuts consist of 75% fat by weight, 80% of which is MUFA (palmitoleic acid). 2. To examine variations in serum lipid levels in response to a high-MUFA diet based on macadamia nuts, 3 week interventions of macadamia nuts, coconuts and butter were determined in young, healthy Japanese female students. 3. After 3 weeks intervention, serum concentrations of total cholesterol and low-density lipoprotein-cholesterol were significantly decreased in the macadamia nut and coconut diets and bodyweight and body mass index were decreased in the group fed macadamia nuts, although there were no statistically significant changes in the group fed butter.

Adult↗

Distribution of isomeric octadecenoic fatty acids in milk fat.

Milk fat octadecenoic acids were isolated by preparative gas-liquid chromatography of their methyl esters and separated into cis and trans isomers by argentation thin-layer chromatography. The distribution of the double bonds in the two fractions was determined by reductive ozonolysis. Cis octadecenoic acids had double bonds in positions 6 to 14. The cis-9 isomer was the most abundant isomer representing approximately 95% of the total. Trans octadecenoic acids had double bonds in positions 6 to 16, the predominant isomer being trans-11 octadecenoic acid. Data illustrating the range of variation in the distribution of positional isomers of octadecenoic acid for herd milk fat and butter are presented. There was no obvious seasonal variation in the distribution pattern. Fat from perirenal and subcutaneous adipose tissue of a dairy cow had composition similar to that of butter samples.

Adipose Tissue↗

Measurement of estrogens in cow's milk, human milk, and dairy products.

Free natural estrogens in raw and commercial whole milk were quantitated by radioimmunoassay. The ranges of concentration of estrone, estradiol 17-beta, and estriol were 34 to 55, 4 to 14, and 9 to 31 pg/ml. Proportions of active estrogens (estrone and estradiol) in the fat phases of milk by radioactive tracer on separated milk were 80% and 65%. These findings were supported by radioimmunoassay of skim milk and butter. Equilibrium dialysis of skim milk with hydrogen 3 labeled estrogens showed that 84 to 85% of estrone and estradiol and 61 to 66% of estriol were protein bound. Whey proteins demonstrated a greater binding capacity than casein. This result was confirmed by radioimmunoassay of dry curd cottage cheese and whey. The concentrations in curd were 35, 11, and 6 pg/g. In whey they were 4, 2, and 3 pg/ml. The quantity of active estrogens in dairy products is too low to demonstrate biological activity. Butter was highest with concentrations of 539, 82, and 87 pg/g. Human colostrum demonstrated a maximum concentration of 4 to 5 ng/ml for estrone and estriol and about .5 ng/ml for estradiol. By the 5th day postpartum, they decreased to become similar to cow's milk.

Animals↗