Nitrosamines in Malt and Malt Beverages

National and local surveys of barley malt and barley malt-containing beverages for volatile N-nitrosamines were conducted by the Food & Drug Administration. Sixty-four samples of locally purchased beers had an average N-nitrosodimethylamine (NDMA) content of 3 ppb. Eleven of 44 Scotch whiskeys had nondetectable NDMA levels. A national sampling of domestic and imported malts revealed that 84% had NDMA concentrations of less than 10 ppb. Domestic and imported beers were also surveyed in a compliance program; 260 beer samples (180 domestic, 80 import) were analyzed. The domestic beers averaged less than 1 ppb and the imported beers averaged 1 ppb. These data indicate that the levels of NDMA in beer have been reduced significantly.     

Copper, zinc, calcium and magnesium content of alcoholic beverages and by-products from Spain: nutritional supply.

Levels of copper, zinc, calcium and magnesium were measured in alcoholic beverages (whiskies, gins, rums, liquors, brandies, wines and beers) and by-products (non-alcoholic liquors and vinegars) using flame atomic absorption spectrometry (FAAS). Mineral concentrations were found to be significantly different between the nine alcoholic and non-alcoholic by-products studied (p < 0.001). In distilled alcoholic beverages, concentrations measured in rums and brandies were statistically lower than those determined in gins and alcoholic liquors (p = 0.001). For Cu, measured concentrations were statistically different for each of the five groups of distilled alcoholic beverages studied (p < 0.001). In fermented beverages, Zn, Ca and Mg levels were significantly higher than those concentrations determined in distilled drinks (p < 0.005). Contrarily, Cu concentrations were statistically lower (p < 0.001). Wines designated as sherry had significantly higher Ca and Mg levels (p < 0.005). White wines had significantly higher Ca and Zn levels (p < 0.05) compared with red wines and, contrarily, Cu concentrations were significantly lower (p < 0.005). In wine samples and corresponding by-products (brandy and vinegar), statistical differences were established for all minerals analysed (p < 0.01). Remarkably, for Cu, the concentrations determined in brandies were statistically higher. On the basis of element levels and the official data on consumption of alcoholic beverages and by-products in Spain, their contribution to the daily dietary intake (DDI) was calculated to be 124.6 microg Cu day(-1) and 193.3 microg Zn day(-1), 40.3 mg Ca day(-1) and 19.9 mg Mg day(-1). From all studied elements, Cu was the one for which alcoholic beverages constitute a significant source (more than 10% of recommended daily intake). These findings are of potential use to food composition tables.     

N-nitrosodimethylamine analysis in Estonian beer using positive-ion chemical ionization with gas chromatography mass spectrometry

N-nitrosodimethylamine (NDMA) is a potent animal carcinogen that has been detected in trace levels in beers. A total of 264 beer samples were analyzed for their NDMA content. For cleaning of the sample the two-step solid-phase extraction with Extrelut and Florisil sorbents were used. NDMA was separated by gas chromatography and detected by positive-ion chemical ionization using ammonia as reagent gas. The HP 6890 Plus GC/HP 5973 MSD with positive-ion chemical ionization option was used in the selected ion-monitoring mode. The limit of detection for NDMA using this method was 0.15 ppb with about 70–80% recovery. Of 158 Estonian beers analyzed during 2003–2004, the average NDMA level was found to be 0.20 ppb. Of 106 imported beer samples the average NDMA level was found to be 0.21 ppb.     

Content of biogenic amines and polyamines in beers from the Czech Republic

Altogether 114 samples of alcoholic and non‐alcoholic bottled beer produced in 28 breweries in the Czech Republic were monitored for levels of biogenic amines (BA) and polyamines (PA). The beers were analysed immediately after purchase and then at the end of the best‐before period (storage temperature 8 ± 1 °C). The concentrations of histamine, phenylethylamine and tryptamine in the studied samples were very low (mostly under 30 mg L^?1). The studied PA spermine and spermidine also occurred in small amounts. Nevertheless, the levels of tyramine, putrescine and cadaverine reached significant values, especially in alcoholic beers. In almost 25% of the tested samples of alcoholic beers (at the end of the best‐before period), the total amount of all the monitored BA and PA exceeded the ‘healthy’ level of 100 mg L^?1, which is considered toxicologically significant, especially in alcoholic beverages. Copyright © 2012 The Institute of Brewing & Distilling     

An investigation of the levels of N-nitrosodimethylamine, apparent total N-nitroso compounds and nitrate in beer

Over 170 retail samples of beer have been analysed for N-nitrosodimethylamine (NDMA), apparent total N-nitroso compounds (ATNC) and nitrate. Levels of NDMA ranged from below 0.1 up to 1.2 micrograms/kg with a mean of 0.2 micrograms/kg. ATNC was detected in 42% of the samples in concentrations of up to 569 micrograms (N-NO)/kg. The levels of nitrate ranged from less than 0.2 up to 143 mg/kg with a mean of 16.8 mg/kg. There was no correlation between the amounts of NDMA and ATNC found in the retail beers. Samples taken during the course of fermentation showed that NDMA was unaffected by the bacterial reduction of nitrate which causes ATNC formation. HPLC studies using a photolysis/chemiluminescence detector revealed that the ATNC in beer are highly polar species of as yet unknown identity.     

Current incidence of N-nitrosodimethylamine in beers worldwide

A range of beers (n = 138) from 42 countries were analysed for the presence of N-nitrosodimethylamine (NDMA) using a chemiluminescence technique with a limit of quantification of 0.1 µg L^-1. The overwhelming majority of samples (79%) did not contain detectable NDMA, and only three samples exceeded 0.5 µg L^-1. No association was found between NDMA content and beer strength, type or geographical origin. It was noted that water can be a potential source of NDMA in beverages.     

Influence of aflatoxin in Nuruk on the safety of starch-based alcoholic beverage

Nuruk is a fermentation agent used to manufacture alcoholic beverages, which contains a variety of microorganisms. Most microorganisms in Nuruk are useful for the production of alcoholic beverages; however, Nuruk can be infected with Aspergillus flavus, which produces aflatoxin (AF). Therefore, this study analyzed total AF concentrations in Nuruk, the transfer of AF from Nuruk to alcoholic beverages, and AF-producing microorganisms to determine the safety of alcoholic beverages with respect to this toxin. ELISA showed that total AF levels in 14 of 61 Nuruk samples exceeded 15 ppb, the Korean permissible level in cereal products. In alcoholic beverages produced with Nuruk containing AF levels >15 ppb, only AF G₁ was detected, at a level of 0.3 ppb, and the transfer ratio of AF G₁ was approximately 1.2% to 1.3%. The dominant genera in Nuruk were Aspergillus and Rhizopus. Among 30 strains belonging to Aspergillus, 10 produced only AF B₁ at levels of 0.1 to 2.4 ppb after incubation at 25 °C for 8 days on potato dextrose agar plates. Although AF in Nuruk was rarely transferred to alcoholic beverages and the aflatoxigenic strains were found to possess poor AF-producing capacity, further efforts to reduce AF in Nuruk are needed to ensure the safety of alcoholic beverages.     

Ochratoxin A in sultanas from Turkey I: Survey of unprocessed sultanas from vineyards and packing-houses.

A method for the determination of ochratoxin A (OTA) in sultanas from Turkey using extraction with a sodium bicarbonate solution (2% NaHCO3) followed by immunoaffinity clean-up and liquid chromatography with fluorescence detection was used to assess the frequency of occurrence and level of OTA. In-house validation was carried out with spiked samples at levels of 0.15, 1.5, 5.0 and 10 microg kg-1 and average recoveries were 91, 93, 87 and 89%, respectively. The limits of detection and limit of quantification in Turkish sultanas were 0.026 and 0.09 microg kg-1, respectively. A survey for the presence of OTA was carried out on 264 unprocessed sultana samples during the production seasons between 1998 and 2000 collected annually from vineyards and from packing-houses. The analyses of unprocessed sultanas showed that 32.2% of the total number of samples contained no detectable OTA, whereas 9.8% of sultana samples had OTA concentrations above 10 microg kg-1, and the remaining 58% had levels within the range 0.026-10 microg kg-1. There were big differences in median concentrations between years. Considering the year of production, it appears that sultanas produced in 1998 and 2000 showed the lowest incidence of OTA contamination (median<0.02 microg kg-1), whereas 2002 showed the highest incidence (median=4.3 microg kg-1). The overall mean OTA concentration was calculated as 3.4 microg kg-1, and the overall median as 0.9 microg kg-1. Among the samples analysed, the highest detected level of OTA was 54 microg kg-1.     

Comparison of ochratoxin A and deoxynivalenol in organically and conventionally produced beers sold on the Belgian market.

Beer was chosen as a cereal-derived and homogeneous product for a comparison of organic and conventional production methods in terms of mycotoxin contamination levels. Ochratoxin A (OTA, a storage mycotoxin) and deoxynivalenol (DON, a field mycotoxin) were assessed by HPLC in organically and conventionally produced beers sold in Belgium. Immunoaffinity column (OchraTest and DONPrep) purification was used prior to HPLC analysis. For in-house validation, recovery experiments, carried out with the spiked beers in the ranges of 50-200 ng OTA l-1 and 20-100 microg DON l-1, led to the overall averages of 91% (RSD = 10%, n = 9) and 93% (RSD = 5%, n = 27), respectively. Organic beers collected during 2003-2004 were more frequently OTA-contaminated (95%, n = 40) than their conventional counterparts (50%, n = 40). Conventional beers were OTA-contaminated at a mean concentration of 25 ng l-1 (range: 19-198 ng l-1), while organic beers contained a mean level of 182 ng l-1 (range: 18-1134 ng l-1). High OTA contamination above the limit of 200 ng l-1 (up to 1134 ng l-1) occasionally occurred in organically produced beers. A complementary survey performed with the same brands in 2005 did not confirm this accidental presence of excessive OTA loads (range: 3-67 ng l-1 for 10 conventional beers and 19-158 ng l-1 for 10 organic beers). Establishing a maximum of 3 microg OTA kg-1 in malt, the application of the regulation EC No. 466/2001 (entered in force before the last sampling) may be related to the observed improvement. The overall incidence of DON was 67 and 80% in conventional and organic beers, respectively. DON concentrations ranged from 2 to 22 microg DON l-1 (mean = 6 microg DON l-1) in conventional beers, while organic beers ranged from 2 to 14 microg DON l-1 (mean=4 microg DON l-1). Thus, DON in beers does not appear to be a major matter of concern. From the statistical tests, it was concluded that the variation between different batches was significant (P < 0.0001), in contrast to that observed between different brands, showing a lack of homogeneity in the raw materials. This occurs either in organically or in conventionally produced materials. Considering these results, an optimized frequency of controls according to European Regulations EC No 466/2001 and EC No 856/2005 should be recommended to reject the irregular batches.     

Multi‐element characterisation of carob,fig and almond liqueurs by MP‐AES

Carob pod, fig and almond liqueurs are produced in various Mediterranean regions. This work reports the mineral content of these traditional beverages and evaluates the influence of the raw material. Twenty‐five fruit liqueurs from 16 producers were analysed. A simple open‐vessel sample mineralization by wet digestion using nitric acid–hydrogen peroxide (1:1) was used before spectrometric analysis. Nine essential elements (Cu, Ca, Mg, Na, K, Fe, Zn, Mn and P) and two non‐essentials (Cd and Pb) were quantified by microwave plasma–atomic emission spectrometry (MP‐AES). Carob liqueur presented the broader profile of minerals and was the only fruit liqueur containing iron (72.7% of samples), phosphorous, manganese and low levels of lead (in two of the 11 samples). Conversely, almond liqueurs contained the lowest mineral content with only five elements detected. Fruit liqueurs showed variability in mineral content even within the same liqueur, presumably as a consequence of the different manufacturing processes. Despite this variability, application of principal component analysis to essential mineral concentrations (K, Na, Ca, Mg, Mn, Fe and Zn) resulted in satisfactory classification (PC1 and PC2 accounting for 78.5% of the total variance) of the Portuguese liqueurs evaluated. Copyright © 2018 The Institute of Brewing & Distilling     

DETERMINATION OF TYRAMINE IN ALCOHOLIC AND NONALCOHOLIC BEERS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH ELECTROCHEMICAL DETECTION

A simple method for the isolation of tyramine from alcoholic and nonalcoholic beers and its subsequent analysis by HPLC with electrochemical detection has been developed. Ultrafiltration of the beer followed by a cation-exchange step provided clear resolution of the tyramine peak and greatly shortened the assay time. The concentration of tyramine in the alcoholic beers ranged from 0.98 to 3.21 mg L⁻¹ and in the nonalcoholic beers from 0.52 to 3.96mg L⁻¹. The possible hazard of consumption of both types of beer to patients on monoamine oxidase inhibitor treatment is discussed. The application of the method to the determination in other beverages is also demonstrated.     

Determination of green,blue and yellow artificial food colorants and their abuse in herb-coloured green Easter beers on tap

Beer is one of the most popular alcoholic beverages worldwide. For consumer acceptance, significant factors are its taste, flavour and colour. This study determines selected synthetic green, blue and yellow food colorants in popular Easter herb-coloured green beers on tap produced in breweries on Holy Thursday. The abuse of beer colouring with Tartrazine (E 102), Quinoline yellow (E 104), Sunset yellow (E 110), Patent blue (E 131), Indigo carmine (E 132), Brilliant blue FCF (E 133), Green S (E 142) and Fast green FCF (E 143) was assessed in 11 green beer samples purchased in local restaurants. HPLC was used for the separation and detection of artificial colorants with diode-array detection and a Chromolith Performance CN 100 × 4.6 mm column with guard pre-column Chromolith CN 5 × 4.6 mm. Separation was performed in gradient elution with mobile phase containing methanol–aqueous 2% ammonium acetate at pH 7.0. The study showed that eight beers (70%) marketed in the Czech Republic contained artificial colorants (Tartrazine and Brilliant blue FCF). The concentration of colorants found in analysed green herb-coloured beers ranged from 1.58 to 3.49 mg l^–1 for Tartrazine, 0.45–2.18 mg l^–1 for Brilliant blue, while Indigo carmine was detected only once at concentration 2.36 mg l^–1. Only three beers showed no addition of the synthetic colorants. However, the levels of artificial colorants found in beers marketed in the Czech region were very low and did not show a serious risk for consumers’ health.     

Application of Gamma Irradiation on Breakdown of Hazardous Volatile N-Nitrosamines

ABSTRACT: Gamma irradiation was applied for the breakdown of the volatile N-nitrosamines (VNAs), nitrosodimethylamine (NDMA), and nitrosopyrrolidine (NPYR). NDMA and NPYR were dissolved in distilled water, dichloromethane, or ethanol, and irradiated at 2.5, 5.0, 7.5, 10, 15, 20, and 25 kGy by gamma ray. Samples were analyzed by GC-TEA. NDMA and NPYR in distilled water and dichloromethane were easily broken at 5 kGy or above. NPYR was the most sensitive to irradiation, while the NDMA is relatively resistant to irradiation. NDMA and NPYR dissolved in ethanol was the most resistant to irradiation and the breakdown was 90% or above at 20 kGy. VNA formation was not observed by in vitro test from decomposed compounds without nitrite, while the VNAs were found by exposure of irradiated products to nitrite. Results indicated that the gamma irradiation technology may be effective to reduce carcinogenic VNA contents in food and other industry.     

Metal profile of traditional alcoholic beverages prepared by the ethnic communities of Assam,India

The rice‐based traditional alcoholic beverages prepared by the ethnic communities, viz., Deori, Mising and Ahom communities of Sivasagar district of Assam, India are known as Sujen, Apong and Xajpani , respectively. The concentrations of Na, K, Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd and Pb in these traditional drinks have been determined with a view to making the general public aware of the average daily intake of these metals from the consumption of these rice beers. The metal contents were estimated using atomic spectrometry after digesting the samples with nitric acid and sulphuric acid. The mean concentrations of the metals (mg/L) in the samples were: 20–60 for Na; 300–460 for K; 0.011–0.325 for Mn; 0.709–4.15 for Fe; 0.13–0.313 for Co; 0.28–0.554 for Ni; 0.008–0.074 for Cu; 0.674–3.968 for Zn; 0.02–0.05 for Cd; and 0.48–1.13 for Pb. The presence of Cr could not be detected. Based on the results, traditional alcoholic beverages consumed by the ethnic communities of Assam, India, can be considered safe and can be used as a supplement of essential nutrients such as potassium, iron, manganese and zinc. The metal contents of these rice beers have been compared with those of wheat and barley beers. Copyright © 2017 The Institute of Brewing & Distilling     

Determination of ethyl carbamate in some fermented Korean foods and beverages

Ethyl carbamate has been associated with cancer for several decades. It is mainly found in fermented foods and beverages. In view of the importance of fermented foods in the Korean diet and the significant level of ethyl carbamate expected, we determined ethyl carbamate concentrations in some of the staple food items and estimated the daily intake for the Korean population. Ethyl carbamate in commercial samples of kimchi, soy sauce, vinegar, soybean paste, and alcoholic beverages were determined by gas chromatography-mass spectrometry/selective ion monitoring (GC-MS/SIM). Homemade soy sauce and kimchi were also analysed. The maximum ethyl carbamate concentrations observed were 73 mu g/kg in soy sauce, 7.9 mu g/kg in soybean paste, 2.5 mu g/l in vinegar, 16.2 mu g/kg in kimchi and 15.4 mu/l in Korean traditional alcoholic beverages. Combining these values with the average daily food intake data, we estimated that the maximum daily exposure of Korean population to ethyl carbamate is 2.8 mu g/day, which is not a negligible amount considering the 'virtually safe dose' derived by animal experiment ranges between 1.2 and 4.8 mu g/day. It would be desirable to closely monitor ethyl carbamate levels in Korean foods and to find ways to reduce the daily intake.     

Mycotoxins in South African traditionally brewed beers

Traditionally brewed alcoholic beverages are regularly consumed by most ethnic black South Africans. Maize and barley, both of which are used for producing locally brewed alcoholic beer, are frequently contaminated by mycotoxin-producing moulds. The study was undertaken to investigate whether these toxins are present in raw grains and the traditional beers imbibed by the local black African population. It was established that the raw ingredients (sorghum, sorghum malt grains, maize grits), commercially produced traditional beers (Utshwala and Utshwala special) and home-brewed beers (Umqombotha, Isiqatha, Imfulamfula) were contaminated by bacteria and fungi (both yeasts and moulds). The contaminating moulds were isolated and identified. The contaminated samples were analysed for aflatoxins B 1 , B 2 , G 1 and G 2 , zearalenone, citrinin, deoxynivalenol, and ochratoxin A using a multi-mycotoxin thin-layer chromatography screening method and the toxins were quantified by high-performance liquid chromatography. Grain samples were infected by Aspergillus flavus , A. alliaceus , A. clavatus , Penicillium spp., Rhizopus spp. and Mucor spp. Sorghum malt grain samples contained the toxin zearalenone. No mycotoxin-producing fungi were present in the fermented beers but two of six commercial beer samples contained aflatoxins (200 and 400 μgl -1 ) and 45% (13 of 29) of the home-brewed beers had zearalenone (range 2.6-426 μg l -1 ) and/or ochratoxin A (3-2340 μg l -1 ).     

Bioactive and Sensory Properties of Herbal Spirit Enriched with Cocoa (Theobroma cacao L.) Polyphenolics

In the Southeast European and Mediterranean regions, consumption of various alcoholic beverages in the form of plain and aromatized spirits and liqueurs is an important part of traditional local gastronomy. Especially herbal spirits and liquors are widely spread and consumed. The objective of this study was to evaluate the potential of using cocoa raw materials (cocoa beans, cocoa nibs or cocoa liquor) in order to enhance the bioactive profile of traditional herbal liqueur, and to produce an appealing cocoa-flavoured alcoholic beverage. Therefore, the polyphenolic profile, antioxidative and sensory properties of herbal liquors containing the macerated cocoa raw materials were monitored during 6 months, in order to evaluate the kinetics of cocoa polyphenols release and optimise the maceration procedure for obtaining the best cocoa-enriched herbal spirit. Compared to plain herbal liquor without the additions, the obtained herbal liqueurs exhibited significantly higher content of polyphenolic antioxidants. Among the used cocoa raw materials for the maceration, cocoa nibs were evaluated as the best source of polyphenolic compounds (herbal liqueur containing cocoa nibs, 3,081.82 mg GAE/L), followed by cocoa liquor (1,897.73 mg GAE/L) and cocoa beans (1,358.64 mg GAE/L). Prolonged maceration time (up to 6 months) resulted with an increase in the content of polyphenolic compounds and the antioxidant capacity of herbal liqueurs. Herbal liqueurs containing the cocoa raw materials of bigger particle size (cocoa beans and cocoa nibs) exhibited better sensory properties (overall acceptability and chocolate-like flavour) than herbal liqueur containing cocoa liquor.     

Solid-phase microextraction of N-nitrosodimethylamine in beer

N-nitrosodimethylamine (NDMA) is a highly active carcinogen that has been detected in trace levels in beers. Solid-phase microextraction (SPME) was used to develop a method for the extraction of NDMA from beer using headspace sampling and gas chromatography with mass spectrometry detection. Polydimethylsiloxane/divinylbenzene (PDMS–DVB) fibres were used to evaluate the influence of equilibrium time, ionic strength, extraction time and temperature by means of a factorial design. The method was validated calculating the linearity, reproducibility, limit of detection and limit of quantification. The method was applied to the quantitative analysis of NDMA combining the standard addition method with an internal standard method.     

Fumonisin B1 contamination of home-grown corn in high-risk areas for esophageal and liver cancer in China.

Fumonisin B1 (FB1) is reportedly the causative agent of several animal mycotoxicoses and has etiologically been linked to human oesophageal and liver cancer in certain areas of South Africa and China. To study a possible relationship between exposure to FB1 and human cancer risk, the current status of FB1 contamination in food samples in Huaian and Fusui, where incidences of oesophageal and liver cancer are amongst the highest in China, was investigated. A total of 259 corn samples were collected from individual households in five villages of different townships in Huaian during December 2001 and December 2002, and in four villages of different townships in Fusui during May 2001 and May 2002. Corn samples were also collected from individual households in two villages in Huantai, an area with low incidences of both cancers. Enzyme-linked immunosorbent assays (ELISA) and immunoaffinity-HPLC methods were used for FB1 analysis. In corn samples from Huaian, FB1 was detectable in 95.7% (112/117) of the samples, with an average of 2.84 mg kg-1 (range 0.1-25.5 mg kg-1). FB1 was detected in 83.0% (78/94) of the Fusui samples, with an average of 1.27 mg kg-1 (range 0.1-14.9 mg kg-1), and in 83.3% (40/48) of Huantai samples, with an average of 0.65 mg kg-1 ranging from 0.1 to 5.7 mg kg-1. The level of FB1 in corn samples from Huaian was significantly higher than from Huantai (P < 0.001). In addition, 47 of 112 (42.0%) positive Huaian samples had FB1 level greater than 2.0 mg kg-1, which was significantly higher than 10.0% (4/40) of Huantai samples (P < 0.001). Furthermore, variations were found between samples collected in different years and different villages. The high contamination rates of FB1 found in food from these areas, along with previous reports, suggest a possible contributing role of FB1 in human esophageal- and hepato-carcinogenesis.     

Investigations of Edible Oils for Volatile Nitrosamines

Twenty-one commercial edible oils and five margarine samples were purchased locally and analyzed for nitrosamines (NAs) by a gas-liquid chromatograph interfaced with a Thermal Energy Analyzer. N-nitrosodimethylamine (NDMA), at levels of 0.22–1.01 ppb, was the only volatile apparent NA detected. These levels were found in 7/7 corn, 6/6 olive, 2/2 sunflower, 1/3 soybean, 2/3 undesignated oils, and 4/5 margarine samples. The other three oils and one margarine sample showed no NDMA at the lower limit of 0.1 ppb. These levels of nitrosamine detected in the edible oils tested were much lower than the levels previously reported.