Survey of ethyl carbamate in fermented foods sold in the United Kingdom in 2004

Results are presented of a survey of fermented foods and beverages sold in the United Kingdom for levels of ethyl carbamate (urethane) carried out to expand the range of food types sold in the United Kingdom for which data regarding ethyl carbamate are available. Samples were analyzed by in-house validated methods, which included measurement uncertainty estimates. The samples comprised 75 fermented liquids (beers, wines, fortified wines, spirits, liqueurs, soy sauces, and vinegars) and 25 fermented solid foods (cheeses, yogurts, soybean products, sauerkraut, yeast extract, olives, and Christmas pudding). Ethyl carbamate was not detected in the beers or the cider. Wines contained between 11 and 24 microg/kg and sake between 81 and 164 microg/kg. Fortified wines contained ethyl carbamate at levels between 14 and 60 microg/kg. Only two of five liqueurs contained ethyl carbamate. Most soy sauces and vinegars did not contain ethyl carbamate. No ethyl carbamate was detected in cheeses, yogurts, olives, or soybean-based products. Single samples of sauerkraut, yeast extract, and Christmas pudding contained low levels (29, 41, and 20 microg/kg ethyl carbamate, respectively).     

Association of key foods and beverages with obesity in Australian schoolchildren

OBJECTIVE: To examine the pattern of intake of key foods and beverages of children aged 4-12 years and the association with weight status. DESIGN AND SETTING: A computer-assisted telephone interview was used to determine the intake of fruit, vegetables, packaged snacks, fast foods and sweetened drinks 'yesterday' and 'usually' as reported by parents/guardians of a representative sample of 2184 children from the Barwon South-Western region of Victoria, Australia. RESULTS: Children who consumed >2-3, >3-4 and >4 servings of fruit juice/drinks 'yesterday' were, respectively, 1.7 (95% confidence interval (CI) 1.2-2.2), 1.7 (95% CI 1.2-2.5) and 2.1 (95% CI 1.5-2.9) times more likely to be overweight/obese compared with those who had no servings of fruit juice/drink 'yesterday', adjusted for age, gender and socio-economic status (SES). Further, children who had > or = 3 servings of soft drink 'yesterday' were 2.2 (95% CI 1.3-3.9) times more likely to be overweight/obese compared with those who had no servings of soft drink 'yesterday', adjusted for age, gender and SES. In addition, children who 'usually' drank fruit juice/drinks twice or more per day were 1.7 (95% CI 1.2-2.4) times more likely to be overweight/obese compared with those who drank these beverages once or less per week, adjusted for age, gender and SES. Although fast foods and packaged snacks were regularly eaten, there were no associations between weight status and consumption of these foods. CONCLUSIONS: Intake of sweetened beverages was associated with overweight and obesity in this population of Australian schoolchildren and should be a target for intervention programmes aimed at preventing unhealthy weight gain in children.     

Free galactose content in selected fresh fruits and vegetables and soy beverages

The free galactose content was determined in three soy beverages, and 34 selected fruits and vegetables purchased at different times of the year and/or local markets in British Columbia, Canada. The objective of the work was to provide additional information on the free galactose content of foods to assist individuals with galactosemia in making dietary decisions. Free galactose contents in the selected plant materials ranged from 2.0 +/- 0.1 mg/100 g in red potato to 39.7 +/- 1.9 mg/100 g in red pepper. Different time of the season, variety, and storage of the product affected the free galactose contents in most of the plant materials measured in this study. Free galactose levels in kiwi, green seedless grapes, and bell peppers were found to be higher than previous reports, whereas the amount of free galactose in three varieties of tomatoes was significantly lower than previously reported. An evaluation of the change of galactose in Roma tomatoes during ripening showed that free galactose levels increased linearly over time, and storage at 4 degrees C significantly increased free galactose levels in tomatoes. Soy beverages made from soy protein isolate contained less free galactose (1.3 +/- 0.2 mg /100 g) compared to the samples made from whole soybeans (4.8 +/- 1.9 and 5.3 +/- 1.7 mg/ 100 g). This study provides additional information on the range of free galactose in fruits and vegetables which will allow individuals with galactosemia to make more informed dietary choices.     

The quantitative analysis of thiamin and riboflavin and their respective vitamers in fermented alcoholic beverages

This research aimed to develop a simple and effective method for analyzing thiamin (B(1)), riboflavin (B(2)) and their respective vitamers by high performance liquid chromatography (HPLC) in fermented alcoholic beverages. The method developed here employs a phosphate buffer/methanol gradient elution on a single reverse phase column, coupled with independent fluorescent detection regimes. It also employs a precolumn derivatization to convert thiamin to thiochrome via an alkaline potassium ferricyanide solution. The method described here allowed a spike recovery of better than 97%, with a typical linear detection range (R(2) ≥ 0.9997) between ≤ 5 and ≥ 500 μg/L for all vitamers studied. Lager style beers were found to contain significantly (p < 0.001) less thiamin than other tested styles of beers (lager, 35.7 μg/L; ale, 88.3 μg/L; stout/porters, 104.4 μg/L; wheat beers, 130.7 μg/L), which may be due to the raw material and extensive processing that occurs for this style. There was no statistical difference (p = 0.608) between the riboflavin content of each beer style. Furthermore, wines and ciders contain less thiamin and riboflavin than beer, which is also likely to be due to the base materials used and the differences in processing steps to produce these beverages.     

High-performance liquid Chromatography/Mass spectrometry analysis of proanthocyanidins in foods and beverages.

Monomeric and oligomeric proanthocyanidins present in a range of plant-derived foods and beverages were separated by degree of polymerization and identified using a modified normal-phase high-performance liquid chromatography (HPLC) method coupled with on-line mass spectrometry (MS) analysis using an atmospheric pressure ionization electrospray chamber. In addition, ultraviolet (UV) and fluorescence detection were used to monitor the separation of proanthocyanidins, with fluorescence detection demonstrating both increased sensitivity and the ability to reduce interfering signals from other components present in the food and beverage matrices as compared to UV detection. This qualitative study demonstrates the ability of this HPLC/MS technique to separate singly and doubly linked procyanidins, prodelphinidins, and copolymer oligomers, including their galloylated derivatives, present in a range of food and beverage samples.     

Lignan content of selected foods from Japan

Lignans constitute a group of phytochemicals widely distributed in the human diet. Once ingested, most dietary lignans are metabolized by the gut microflora to enterolactone and enterodiol, also known as enterolignans. Together with isoflavones and coumestrol, enterolignans have been traditionally classified as phytoestrogens, plant-derived compounds that exhibit estrogen-like activity. In addition to a higher intake of vegetables, many studies have suggested that the presence of phytoestrogens in the Japanese diet is one of the factors that might explain the reduced incidence of certain chronic diseases in Japanese populations compared with Western countries. Being a vegetable-based diet, exposure to dietary lignans should be high, but to assess this exposure, a lignan food database is required. Stable isotope-dilution gas chromatography coupled with mass spectrometry was used to quantify six plant lignans in 86 food items commonly consumed in Japan. These data will complement the previous databases and most importantly expand the knowledge of occurrence of lignans in food to Eastern diets.     

Contents of anthocyanins and ellagitannins in selected foods consumed in Finland

Numerous in vitro and in vivo studies have suggested that dietary anthocyanins and ellagitannins or ellagic acid might have beneficial health effects. Epidemiological evidence on the disease-preventing potential of these polyphenols is lacking, due to the absence of reliable data on their contents in foods. In this study was analyzed the content of anthocyanins and ellagitannins (as ellagic acid equivalents after acid hydrolysis) in foods consumed in Finland, including berries, fruits, vegetables, and processed products, using high-performance liquid chromatographic (HPLC) methods. Anthocyanins were detected in 41 of 54 selected food items. The total anthocyanin content varied in berries from 1 to 611 mg/100 g, in fruits from 2 to 66 mg/100 g, and in vegetables from 3 to 75 mg/100 g of fresh weight as the weight of the aglycone. Ellagitannins were screened in 33 food items, but were detected only in 5 species of berries, that is, in cloudberry, raspberry, rose hip, strawberry, and sea buckthorn, the content ranging from 1 to 330 mg/100 g. The results underscore the superiority of berries, especially dark blue or red berries, as excellent sources of anthocyanins and certain berries of the Rosaceae family as the major source of ellagitannins in the Finnish diet.     

Rapid gas chromatographic method for determining ethyl carbamate in alcoholic beverages with thermal energy analyzer detection

A rapid column elution method has been developed for the determination of ethyl carbamate (EC) in alcoholic beverages. The beverage is mixed with Celite and packed in a column containing deactivated alumina capped with a layer of sodium sulfate. EC is then eluted with methylene chloride. The method, using a gas chromatograph-thermal energy analyzer with a nitrogen converter for detection and quantitation of EC, has been applied to a variety of alcoholic beverages. Recoveries +/- standard deviations of EC in wine and whisky fortified at the 20 and 133 micrograms/kg (ppb) levels averaged 87.3 +/- 5.3 and 88.7 +/- 3.6%, respectively. The method has a limit of detection of 1.5 ppb. Gas chromatography/mass spectrometry/mass spectrometry was used to confirm the identity and quantitation of EC in selected beverage extracts.     

The 'Sydney Principles' for reducing the commercial promotion of foods and beverages to children

A set of seven principles (the 'Sydney Principles') was developed by an International Obesity Taskforce (IOTF) Working Group to guide action on changing food and beverage marketing practices that target children. The aim of the present communication is to present the Sydney Principles and report on feedback received from a global consultation (November 2006 to April 2007) on the Principles. The Principles state that actions to reduce marketing to children should: (i) support the rights of children; (ii) afford substantial protection to children; (iii) be statutory in nature; (iv) take a wide definition of commercial promotions; (v) guarantee commercial-free childhood settings; (vi) include cross-border media; and (vii) be evaluated, monitored and enforced. The draft principles were widely disseminated and 220 responses were received from professional and scientific associations, consumer bodies, industry bodies, health professionals and others. There was virtually universal agreement on the need to have a set of principles to guide action in this contentious area of marketing to children. Apart from industry opposition to the third principle calling for a statutory approach and several comments about the implementation challenges, there was strong support for each of the Sydney Principles. Feedback on two specific issues of contention related to the age range to which restrictions should apply (most nominating age 16 or 18 years) and the types of products to be included (31% nominating all products, 24% all food and beverages, and 45% energy-dense, nutrient-poor foods and beverages). The Sydney Principles, which took a children's rights-based approach, should be used to benchmark action to reduce marketing to children. The age definition for a child and the types of products which should have marketing restrictions may better suit a risk-based approach at this stage. The Sydney Principles should guide the formation of an International Code on Food and Beverage Marketing to Children.     

Chemical conversion of alpha-keto acids in relation to flavor formation in fermented foods

Formation of flavor compounds from branched-chain alpha-keto acids in fermented foods such as cheese is believed to be mainly an enzymatic process, while the conversion of phenyl pyruvic acid, which is derived from phenylalanine, also proceeds chemically. In this research, the chemical conversion of alpha-keto acids to aldehydes with strong flavor characteristics was studied, with the main focus on the conversion of alpha-ketoisocaproic acid to the aldehyde 2-methylpropanal, and a manganese-catalyzed reaction mechanism is proposed for this conversion. The mechanism involves keto-enol tautomerism, enabling molecular oxygen to react with the beta-carbon atom of the alpha-keto acid, resulting in a peroxide. This peroxide can react in several ways, leading to unstable dioxylactone or noncyclic intermediates. These intermediates will break down into an aldehyde and oxalate or carbon oxides (CO and CO(2)). All the alpha-keto acids tested were converted at pH 5.5 and in the presence of manganese, although their conversion rates were rather diverse. This chemical reaction might provide new ways for controlling cheese flavor formation with the aim of acceleration of the ripening process or diversification of the flavor characteristics.     

Tocopherol and tocotrienol levels of foods consumed in Hawaii

Because of the individual biological effects and the uncertain or missing information on levels of tocopherols (T) and tocotrienols (T3) in foods frequently consumed in Hawaii, 79 food items (50 in duplicate) were analyzed for alpha-, beta-, gamma-, and delta-tocopherol (alphaT, betaT, gammaT, and deltaT) and alpha-, beta-, gamma-, and delta-tocotrienol (alphaT3, betaT3, gammaT3, and deltaT3) in addition to alpha-tocopheryl acetate (alphaTac). Foods from local markets were stored according to usual household habits, freeze-dried, homogenized, and extracted three times with hexane containing butylated hydroxytoluene as a preservative and tocol as an internal standard. A normal-phase high-pressure liquid chromatography system was applied with fluorescence and photodiode array detection that resulted in baseline separation of all eight analytes and the internal standard tocol (To). The sum of all E vitamer concentrations, or total E vitamers (TEV), in all foods analyzed ranged an average from 0.6 to 828 mg/kg (T < or = 542 mg/kg and T3 < or = 432 mg/kg) and showed the following ranges: oils, 497-828 mg/kg (mainly alphaT and gammaT); margarines, 359-457 mg/kg (mainly gammaT); salad dressings, 20-291 mg/kg (mainly gammaT, except alphaT when soy oil was the main ingredient); cookies, 54-138 mg/kg (mainly gammaT); snacks, 101-220 mg/kg (mainly gammaT); nuts, 22-201 mg/kg (mainly alphaT); vegetables, 2-152 mg/kg (mainly alphaT); pasta, 24-90 mg/kg; cereals, 4-56 mg/kg (mainly betaT3 followed by alphaT); fish, 2-39 mg/kg (mainly alphaT); fried tofu, 64 mg/kg (mainly gammaT); breads, 20-22 mg/kg (mainly betaT3); fat-free mayonnaise, 5 mg/kg (mainly alphaT); poi (fermented taro root), 2 mg/kg (mostly alphaT); and fruits, 2 (papaya) to 13 mg/kg (canned pumpkin) with alphaT predominating. Cereals fortified with alphaTac ranked third and eighth among all foods assayed regarding alphaT and TEV levels, respectively. As compared to the few data available in the literature, our values agreed with some (corn flakes, mango fruit, fat-free mayonnaise, dry-roasted macadamia nuts, dry-roasted peanuts, mixed nuts, spaghetti/marinara pasta sauce, oils, and red bell pepper) but differed for many other items. Our results provide new information on the E vitamer content in foods, emphasize the vast differences of bioactivities of individual E vitamers, and confirm the need for analyses of foods consumed in specific study populations.     

Determination of ethyl carbamate in distilled alcoholic beverages by gas chromatography with flame ionization or mass spectrometric detection

Quantitative methods are detailed for determination of ethyl carbamate in distilled alcoholic beverages by capillary gas chromatography with flame ionization detection (GC/FID) and by packed-column gas chromatography/mass spectrometry (GC/MS) using selected ion monitoring. Five g samples of distillate of known ethanol concentration are diluted with water to 25% ethanol (v/v), washed with petroleum ether, and extracted with dichloromethane prior to GC/FID or GC/MS analysis. As necessary, sample extracts that exhibit GC/FID interference are passed through alumina for additional cleanup. When internal standards (tert-butyl carbamate and n-butyl carbamate for GC/FID, or ethyl 13C-15N-carbamate for GC/MS) were used for quantitation, the limit of detection for ethyl carbamate was in the range of 5-25 ppb. Coefficients of variation ranged from 3.5 to 6.0% for GC/FID determinations, and from 1.4 to 3.2% for GC/MS. Correlation between methods for 22 random distillate samples ranging in concentration from approximately 40 to 800 ppb gave a correlation coefficient (r) of 0.996.     

Available (ileal digestible reactive) lysine in selected pet foods

A recently developed assay for determining available lysine (true ileal digestible reactive lysine) in foods and feedstuffs was applied to 20 commercially available cat foods. Semisynthetic diets, containing cat food as the sole protein source, were prepared. Titanium dioxide was included as an indigestible marker. The diets were fed to growing rats and digesta from the terminal ileum collected and analyzed, along with the diets, for reactive lysine. True reactive lysine digestibility was determined after correction for endogenous lysine loss at the terminal ileum of rats fed an enzyme-hydrolyzed casein-based diet. The amounts of digestible total lysine (conventional method) were also determined. Ileal total lysine digestibility significantly (P<0.05) underestimated (3.6-10.2%) lysine availability (ileal reactive lysine digestibility) for most of the cat foods tested. Ileal digestible total lysine significantly (P<0.05) overestimated the amount of dietary available lysine for all of the cat foods tested by between 38 and 143%. Total lysine digestibility determined using the conventional method of lysine analysis was inaccurate when applied to commercially available cat foods.     

Determination of sulfite in foods and beverages by ion exclusion chromatography with electrochemical detection: collaborative study

A liquid chromatographic (LC) method for determination of total sulfite in foods and beverages by alkali extraction followed by ion exclusion chromatographic separation and electrochemical detection (IEC-EC) was collaboratively studied by 9 laboratories. Blind duplicate samples of starch, diluted lemon juice, wine cooler, dehydrated seafood, and instant mashed potatoes were analyzed without spiking and with added sulfite at 2 levels. The initial sulfite levels varied from 0 to 384 ppm SO2, and the levels added varied from 10 to 400 ppm. The initial sulfite levels determined by the IEC-EC method and the Monier-Williams method were in good agreement. Recovery of added sulfite by the IEC-EC method was generally higher than that by the Monier-Williams method. Within-laboratory repeatability (RSDr) for the IEC-EC method varied from 4.4 to 26.0%, and overall reproducibility (RSDR) varied from 8.5 to 39.3%. The collaborators found the method to be fast, sensitive, and easy to use, which makes it a useful alternative to the Monier-Williams method. The method has been adopted official first action.     

Capillary column gas chromatographic determination of ethyl carbamate in alcoholic beverages with confirmation by gas chromatography/mass spectrometry

A method is described for determining ethyl carbamate at low microgram/kg levels in several types of alcoholic beverages by capillary column gas chromatography with Hall electrolytic conductivity detection and confirmation by mass spectrometry. Samples are diluted to obtain a uniform concentration of ethanol (ca 10%) then saturated with NaCl and extracted with methylene chloride. Extracts are evaporated to a small volume and injected in ethyl acetate solution for chromatographic analysis. The method was evaluated by 5 laboratories, 4 employing the Hall detector and one using mass spectrometric detection. Overall between-laboratory mean percent recoveries were: wine, 85.3 +/- 21.0% coefficient of variation (CV) (spiking level 20-45 micrograms/kg); sherry, 83.8 +/- 16.1% CV (spiking level, 81-142 micrograms/kg); whiskey, 79.5 +/- 13.9% CV (spiking level 127-190 micrograms/kg); and brandy, 85.0 +/- 12.5% CV (spiking level 297-446 micrograms/kg). Mass spectrometric results agreed well with the Hall results for all commodities. Detection limits were about 5 micrograms/kg for the Hall detector and about 0.5 microgram/kg for mass spectrometric detection.     

Gas-liquid chromatographic-mass fragmentographic determination of low levels of diethylcarbonate in beverages.

Sodium chloride and ethanol (omitted for samples with greater than 10% alcohol) are added to the beverage sample and the sample is allowed to equilibrate in a 30 degrees C water bath. An aliquot of the headspace is injected into a gas chromatography containing a column packed with 0.2% Carbowax 1500 on 80--100 mesh Carbopack C. During the elution of diethylcarbonate (DEC), an impurity that is present in diethylpyrocarbonate, the column effluent is vented to a mass spectrometer with a multiple ion detection system and operated in the electron impact mode. The ions at m/e 63 and 91 are monitored. Lemonade, fruit drinks, wine, and beer samples (138 total) were analyzed for DEC. Sixteen samples had greater than 30 ppb DEC. Eight analyses of a lemonade sample gave a mean of 88 ppb with a coefficient of variation of 11%.