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BACKGROUND/OBJECTIVES: An association between biomarkers of trans fat intake and greater risk of preeclampsia has been reported, but research in this area is scant. Thus, we examined the association of second trimester intake of trans fats with risk of preeclampsia and severe preeclampsia. SUBJECTS/METHODS: We followed 67,186 pregnancies of women participating in the Danish National Birth Cohort between 1998 and 2003. Diet was assessed with a food frequency questionnaire at gestation week 25, and preeclampsia diagnosis was obtained by linkage with the Danish National Patient Registry. RESULTS: There were 1804 cases of preeclampsia and 402 cases of severe preeclampsia identified in the cohort. Intake of trans fats decreased during the study period as a consequence of a reduction in industrial trans fat intake. Second trimester intake of trans fats was unrelated to risk of preeclampsia or severe preeclampsia. The relative risk (95% confidence interval; P, trend) of preeclampsia and severe preeclampsia comparing top to bottom quintiles of trans fat intake were 0.95(0.81; 1.11, 0.33) and 1.07 (0.78; 1.48, 0.92), respectively. CONCLUSION: Second trimester intake of trans fats is unrelated to risk of preeclampsia within the intake range observed in a period of gradual reduction of industrial trans fats from the Danish food supply.

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The monitoring of food additives and recent dietary surveys carried out in Denmark have earlier been used to estimate the intake of sweeteners and nitrite in relation to acceptable daily intakes. The ubiquitous use of the preservatives benzoic and sorbic acids raises the question of the magnitude of the intake of these preservatives in relation to acceptable daily intakes. This area is explored in this paper. The content of benzoic and sorbic acids in all food groups, where they are allowed, was monitored in Denmark 17 times between 2001 and 2006 with a total of 1526 samples. Transgressions of maximum limits, illegal use or declaration faults were found in about 3% of samples. From repeated investigations on fat-based foods (salads and dressings), marmalade and stewed fruit, it is concluded that the amounts used in industry have been relatively stable throughout the whole period, although limited data for marmalade show some variation. Most foods in the categories soft drinks, dressings, fat-based salads, pickled herrings, and marmalade contain benzoic and sorbic acid, and sliced bread also contains in some cases sorbic acid. The median daily intake and intake distribution of benzoic and sorbic acids were calculated with data from the Danish National Survey of Dietary Habits and Physical Activity (age from 4 to 75 years) conducted in 2000-2004 with 5785 participants. The median intakes of both benzoic acid and sorbic acid are well below the acceptable daily intakes of 0-5 and 0-25 mg kg(-1) body weight (bw) day(-1) for benzoic and sorbic acid, respectively. However, the 90th percentile based on the average of the samples with a content of benzoic acid is higher than the acceptable daily intake for both men and women, with the highest value of 16 mg kg(-1) bw day(-1) for both boys and girls in the 4-6-year-old age group. Based on the average of all samples, the 95th percentile is over the acceptable daily intake for men up to 34 years and for women up to 24 years, and the 90th percentile for men up to 18 years and for women up to 10 years. Soft drinks, salads and dressings are the main contributors to benzoic acid intake. The sorbic acid intake based on the average of all samples is well below the acceptable daily intake. However, for the intake based on the average of samples with content, the 95th percentile exceeds the acceptable daily intake. This is caused by the dominating contribution to the intake of sorbic acid from sliced bread, but since only seven out of 42 samples have added sorbic acid, the calculation based on the average of samples with content will exaggerate the intake. With a built-in safety factor of 100 in the acceptable daily intakes and judging from the literature, the high intakes of benzoic acid should not cause any concern for ill-effects. However, there must be a reason to reconsider the maximum limits especially for benzoic acid in soft drinks, dressings and salads and for sorbic acid in sliced bread.

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The content of nitrite and nitrate in cured meat products has been monitored in Denmark seven times between 1995 and 2006. The maximum permitted added amounts of sodium nitrite in Denmark (60 mg kg(-1) for most products up to 150 mg kg(-1) for special products) have not been exceeded, except for a few samples back in 2002. The intake, mean and intake distribution of sodium nitrite have been calculated from 1998 to 2006 with data from the Danish dietary survey conducted in 2000-02 on Danes from four to 75 years of age. The amounts used by industry have been relatively stable through the whole period with levels varying between 6 and 20 mg sodium nitrite kg(-1) with sausages, meat for open sandwiches and salami-type sausages being the greatest contributors. The mean intake of sodium nitrate was around 1 mg day(-1), which is very low compared with the total intake of 61 mg day(-1). The mean intake of sodium nitrite was 0.017 and 0.014, 0.009 and 0.008, and 0.007 and 0.003 mg kg(-1) body weight day(-1) for men and women in the age groups 4-5, 6-14 and 15-75 years, respectively, which was much lower than the acceptable daily intake (ADI) of 0.09 mg kg(-1) body weight day(-1). The 99th percentile for the group of 4-year-olds was 0.107 and 0.123 mg kg(-1) body weight day(-1) for boys and girls, respectively, and the 95th percentile was 0.057 and 0.073 mg kg(-1) body weight day(-1) for boys and girls, respectively, highest for the girls. With fewer than 100 boys and girls in the 4-5-year age group, only very few persons were responsible for the high intake. The conversion of nitrate to nitrite in the saliva and the degradation of nitrite during production and storage must also be considered when evaluating the intake of nitrite.

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In 2005, 76 out of 177 analysed samples of non-alcoholic beverages were found to contain the intense sweeteners cyclamate, acesulfame-K, aspartame, and saccharin. The content of cyclamate did not exceed the now permitted maximum level in the European Union of 250 mg l(-1) in soft drinks. The estimated intake of the sweeteners was calculated using the Danish Dietary Survey based on 3098 persons aged 1-80 years. The estimated intake with 90th percentiles of 0.7, 0.8 and 0.2 mg kg(-1) body weight day(-1) for acesulfame-K, aspartame, and saccharin, respectively, was much lower than the acceptable daily intake values of 15, 40, 7, and 2.5 mg kg(-1) body weight day(-1) for acesulfame-K, aspartame, and saccharin, respectively, and on the same level as in the similar investigation from 1999. In contrast to the 1999 investigation, the 90th percentile of the estimated cyclamate intake in 1-3 year olds with 3.7 mg kg(-1) body weight day(-1) was in 2005 lower than the acceptable daily intake of 7 mg kg(-1) body weight day(-1). However, the 99th percentile for 1-3 year olds with 7.4 mg kg(-1) body weight day(-1) still exceeded the acceptable daily intake slightly. The 90th percentile for the whole population with 0.9 mg kg(-1) body weight day(-1) was halved compared with 1999. The reduction in the European Union of the maximum permitted level for cyclamate from 400 to 250 mg l(-1) has brought the intake of cyclamate in small children down to well below the acceptable daily intake value.

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In 1999, 116 samples of non-alcoholic beverages were analysed for the intense sweeteners cyclamate, acesulfame-K, aspartame and saccharin. High contents of cyclamate close to the maximum permitted level in 1999 of 400 mg l(-1) were found in many soft drinks. The estimated intake of the sweeteners was calculated using the Danish Dietary Survey based on 3098 persons aged 1-80 years. The estimated intake with 90th percentiles of 0.7, 4.0 and 0.2 mg kg(-1) body weight (bw) day(-1) for acesulfame-K, aspartame and saccharin, respectively, was much lower than the acceptable daily intake (ADI) values of 15, 40 and 2.5 mg kg(-1) bw day(-1) for acesulfame-K, aspartame and saccharin, respectively. However, the 90th percentile of the estimated cyclamate intake in 1-3 year olds was close to the ADI value of 7 mg kg(-1) bw day(-1); and the 99th percentile in the 1-10 year olds far exceeded the ADI value. Boys aged 7-10 years had a significantly higher estimated intake of cyclamate than girls. The 90th percentile for the whole population was 1.8 mg kg(-1) bw day(-1). After the reduction in the maximum permitted level in the European Union in 2004 from 400 to 250 mg cyclamate l-1, the exposure in Denmark can also be expected to be reduced. A new investigation in 2007 should demonstrate whether the problem with high cyclamate intake is now solved.

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OBJECTIVE: To estimate the intake of ruminant trans fatty acids (TFA) in the Danish population aged 1-80 years. DESIGN: Descriptive study. SUBJECTS: A sex- and age-stratified random sample drawn from the Danish Civil Registration System. A total of 3098 participants (51% female) aged 1-80 years were included. The participation was 66%. DIETARY INFORMATION: A 7-day dietary record. RESULTS: The estimated median intake of ruminant TFA was 1.4 g/day with the 80% central range being from 0.9 to 2.1 among children aged 1-6 years and 1.6 g/day (1.0-2.4) among children aged 7-14 years. The median TFA intake was 1.8 g/day (0.9-2.9) among adults aged 15-29 years and among adults aged 30-80 years. The intake expressed as percentage of energy intake was 0.8, 0.6, 0.7, and 0.7, respectively. Dairy products were the main source of ruminant TFA. CONCLUSIONS: The median intake of ruminant TFA in the Danish population aged 1-80 years is estimated to be 1.7 g/day (0.9-2.7), corresponding to 0.7% of energy intake (0.5-1.0), with dairy products being the main source of ruminant TFA. SPONSORSHIP: The Danish Heart Foundation (Grants 02-2-9-8-22010 and 03-2-9-4-22087) and the Female Researchers in Joint Action (FREJA) programme from the Danish Medical Research Council.

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Degradation of folic acid may occur during extraction of multivitamin-mineral preparations. The degradation may be caused by presence of ions such as Fe3+ and Cu2+, however, the buffer composition may also be critical. This study presents an optimised extraction procedure tested on 24 different products of multivitamin-mineral tablets. The present method yielded mean recoveries of 97% (n = 20) for folic acid and prevented degradation of folic acid in at least 24 h in extracts from multivitamin-mineral tablets.

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OBJECTIVE: To assess the intake of trans fatty acids (TFA) and other fatty acids in 14 Western European countries. DESIGN AND SUBJECTS: A maximum of 100 foods per country were sampled and centrally analysed. Each country calculated the intake of individual trans and other fatty acids, clusters of fatty acids and total fat in adults and/or the total population using the best available national food consumption data set. RESULTS: A wide variation was observed in the intake of total fat and (clusters) of fatty acids in absolute amounts. The variation in proportion of energy derived from total fat and from clusters of fatty acids was less. Only in Finland, Italy, Norway and Portugal total fat did provide on average less than 35% of energy intake. Saturated fatty acids (SFA) provided on average between 10% and 19% of total energy intake, with the lowest contribution in most Mediterranean countries. TFA intake ranged from 0.5% (Greece, Italy) to 2.1% (Iceland) of energy intake among men and from 0.8% (Greece) to 1.9% among women (Iceland) (1.2-6.7 g/d and 1.7-4.1 g/d, respectively). The TFA intake was lowest in Mediterranean countries (0.5-0.8 en%) but was also below 1% of energy in Finland and Germany. Moderate intakes were seen in Belgium, The Netherlands, Norway and UK and highest intake in Iceland. Trans isomers of C18:1 were the most TFA in the diet. Monounsaturated fatty acids contributed 9-12% of mean daily energy intake (except for Greece, nearly 18%) and polyunsaturated fatty acids 3-7%. CONCLUSION: The current intake of TFA in most Western European countries does not appear to be a reason for major concern. In several countries a considerable proportion of energy was derived from SFA. It would therefore be prudent to reduce intake of all cholesterol-raising fatty acids, TFA included.

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A high-performance liquid chromatographic (HPLC) separation method with photo-diode array (PDA) and mass spectrometric (MS) detection was developed to determine and quantify flavonols, flavones, and flavanones in fruits, vegetables and beverages. The compounds were analysed as aglycones, obtained after acid hydrolysis of freeze-dried food material. Identification was based on retention time, UV and mass spectra by comparison with commercial standards, and the UV peak areas were used for quantitation of the flavonoid contents. Examples of HPLC-MS, analyses of orange pulp, tomato, and apple are presented. The method has been used to screen foods on the Danish market, and the contents of flavones, flavonols, and flavanones were measured.

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To estimate the contents of flavonoids in the Danish diet, we have used a high performance liquid chromatographic method for the analysis of fruits, vegetables and beverages. The flavonoid contents were quantified, and the observations were verified by mass spectrometry. The investigation is not completed, but we are able to give an preliminary estimate of the daily flavonoid intake at 26 mg/day. The major food contributors are tea, onions, apples, oranges, and orange juice. Other food subjects have high flavonoid contents, but the daily intake is low, so their contributions to the average daily intake are negligible.

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Several non-nutritive components in fruits, vegetables, herbs and spices have been found to inhibit tumour formation in experimental animals exposed to carcinogens. The active non-nutritive components vary with respect to their chemical structures, and may be classed as phenols, terpenes, indoles, isothiocyanates, allyl sulphides or others. They also seem to work by different mechanisms, being inducers or inhibitors of various enzymes, antioxidants, scavengers of reactive metabolites, or inducers of apoptosis. The dietary levels are generally in the order of 1-100 mg/day for most classes of compounds in the Danish population, and similar levels are expected in most northern European countries. These levels are very low compared with the levels used in most animal experiments, where non-nutritive factors have individually been shown to have inhibitory actions on tumorigenesis. Human long-term intervention trials with antioxidants have generally been discouraging. In human short-term intervention studies, where increased dietary levels of specific vegetables or fruits are studied, doses are also comparatively low. Effects on important enzymes have been reported in several such studies, indicating that low levels of non-nutritive factors could influence carcinogenesis by specific mechanisms. Meta-analyses of cohort studies on specific food items rich in specific non-nutritive components, indicate that carotenoid- or glucosinolate-rich foods protect against some cancers, while flavonoid rich food items do not uniformly show protective effects.

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Breast milk was treated with (1) conventional heating (in water bath) vs microwave heating; (2) microwave heating at two power levels (30% and 100%); (3) increasing final temperatures; and (4) microwave thawing vs refrigerator thawing and examined for changes in specific immunoglobulins to a pool of E. coli and poliovirus type 1 antigens, vitamins E and B1, and the polyunsaturated fatty acids linoleic and linolenic acid. Immunoglobulin activities were stable until final milk temperatures of around 60-65 degrees C were reached, and total inactivation occurred at 77 degrees C. Heating even to high final temperatures did not change contents of vitamins and polyunsaturated fatty acids. No differences in immunoglobulins and nutrients were demonstrated between microwave heating and conventional heating, and between power levels or thawing methods. The study shows that microwave heating of human milk can be performed without significant losses of examined immunoglobulins and nutrients, provided that final temperatures are below 60 degrees C.

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Trans fatty acids from hydrogenated vegetable and marine oils could be as hypercholesterolemic and atherogenic as saturated fatty acids. Hence, it is important to know the fatty acid composition in major food contributors, e.g., margarines and shortenings. In 1992 margarines were examined, and in 1995 brands covering the entire Danish market were examined. Significant amounts of trans-18:1 were found only in hard margarines (mean: 4.2 +/- 2.8%) and shortenings (mean: 6.8 +/- 3.1%), whereas the semisoft and soft margarines contained substantially less trans-18:1 in 1995 than in 1992. Where marine oils had been used to a larger degree the mean trans-monoenoic content was about 15%, of which close to 50% was made up of long-chain (C20 and C22) trans fatty acids. A noteworthy decrease in the content of trans-18:1 had occurred for the semisoft margarines, from 9.8 +/- 6.1% in 1992 to 1.2 +/- 2.2% in 1995. Calculated from sales figures, the supply of trans-18:1 plus saturated fatty acids from margarines has decreased over this three-year period by 1.4 g/day, which has been replaced by cis monounsaturated and polyunsaturated fatty acids.

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Twelve healthy young men followed a 10-d controlled diet that included 210 g of fatty fish d-1. The diet was repeated after 18 d, but with lean meat substituted for fish. Blood samples were collected for assessment of serum lipids and haemostatic variables in the plasma. Both experimental diets caused serum triglycerides and plasma factor VIIc to decline to the same extent. The meat diet was also associated with significant changes in plasma levels of tissue plasminogen activator (t-PA) antigen. PA inhibitor type I (PAI-1) antigen, PAI activity, and t-PA activity of the euglobulin fraction of plasma. The fish diet left these variables unchanged from initial values. Thus, in a paired comparison of the two diets, the fish diet was associated with higher levels of t-PA antigen (5.4 vs. 4.7 g ml-1), which is considered to be beneficial with regard to prevention of cardiovascular disease. However, the fish diet was concurrently associated with the putative unfavourable higher levels of PAI-1 antigen (3.0 vs. 1.2 ng ml-1) and PAI activity (6.1 vs. 3.2 IU ml-1), and lower t-PA activity (80 vs. 140 mIU ml-1). Thus it is unclear which of the two diets has the greatest potential in the prevention of cardiovascular disease.

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The biological activity of 2 R, 4' R, 8' R-(RRR)-a-tocopherol and all-rac-a-tocopherol has been determined by 3 different bioassays: Resorption-gestation test, red blood cell hemolysis test and rat liver storage test. Expressed in relation to all-rac-a-tocopheryl acetate is found by the resorption-gestation test with the doses given in vegetable oil potency ratios of 0.79 for all-rac-a-tocopherol and 1.06 for RRR-a-tocopherol, by the red blood cell hemolysis test ratios of 0.85 for all-rac-a-tocopherol and 1.13 for RRR-a-tocopherol and by the rat liver storage test ratios of 1.03 for all-rac-a-tocopherol and 1.32 for RRR-a-tocopherol. Furthermore is by the resorption-gestation test found a potency ratio of 0.90 between RRR-a-tocopheryl succinate and all-rac-a-tocopheryl acetate. By the evaluation of these figures, most weight must be added to the results from the resorption-gestation test, as it involves the most complex biological mechanism of the 3 methods. The potency ratios found in this investigation are significantly lower than the currently accepted potency ratios of 1.10 between all-rac-a-tocopherol and all-rac-a-tocopheryl acetate, 1.49 between RRR-a-tocopherol and all-rac-a-tocopheryl acetate and 1.21 between RRR-a-tocopheryl succinate and all-rac-a-tocopheryl acetate (all values calculated from their corresponding acetate by multiplication with the ratio of the molecular weight), but the potency ratio between RRR-a-tocopherol and all-rac-a-tocopherol calculated from the 3 different bioassays does not deviate significantly from the currently accepted value of 1.36. It seems necessary to reconsider the biological activity of all-rac-a-tocopherol and RRR-a-tocopherol, and it is suggested to use a potency ratio of 0.80 between all-rac-a-tocopherol and all-rac-a-tocopheryl acetate and 1.0 between RRR-a-tocopherol and all-rac-a-tocopheryl acetate.

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