Short communicationInvestigations on the occurrence of mycotoxins in beer
Introduction
Beer is the most consumed alcoholic beverage in the EU (WHO, 2014). In 2013, the average per capita consumption was 71 L in Europe. In some countries, annual consumption is more than 100 L (The Brewers of Europe, 2014). Therefore, the dietary exposure to natural toxins via beer might be of relevance for a high percentage of consumers in Europe.
Traditional raw materials for beer production are barley and wheat. These grains have often been associated with contamination of mycotoxins (Pereira, Fernandes, & Cunha, 2014). In a former review on mycotoxin carry-over from contaminated grains into beer, it has been suggested that existent mycotoxin levels in the raw grain might further increase as a result of promoted fungal growth in the malting process (Scott, 1996). According to previous reports on their moderate stability during food processing (Bullerman and Bianchini, 2007, Milani and Maleki, 2014), mycotoxins may, to some extent, effectively overcome the brewing process and thus be transferred from malt into beer. Therefore, the presence of mycotoxins in beer potentially is a significant source of intake. Research in this field has mainly been focused on the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEA), which are known to frequently contaminate brewing grains (Barthel et al., 2012; Běláková et al., 2014, SCOOP, 2003). The occurrence of these mycotoxins in beer has been examined in several studies (Curtui et al., 2005, Kappenstein et al., 2005, Kuzdraliński et al., 2013, Samar and Resnik, 2002). Since their potential to cause serious health problems in animals and humans has been well documented (Escrivá et al., 2015, Richard, 2007), the European Commission has established a range of maximum limits for DON and ZEA in cereals and cereal-based foods in order to prevent human mycotoxicoses in consequence of consumption of contaminated foodstuffs (EC, 2006). Maximum amounts for these mycotoxins in beer have not specifically been set so far.
Other groups of mycotoxins such as ergot alkaloids and Alternaria toxins have also been related with detrimental effects according to intake of contaminated foods (Asam and Rychlik, 2013, Crews et al., 2009, Dong et al., 1987). Ergot alkaloids, mainly produced by the fungus Claviceps purpurea, have been known since the Middle Ages to cause intoxications in humans and animals, described as ergotism, which is characterized by a variety of symptoms ranging from abdominal pain, vomiting, and neurological signs to the point of severe vasoconstrictive effects (Krska & Crews, 2008). Otherwise, some ergot alkaloids also play an important role as pharmaceuticals for treatment of vascular and neurological diseases, including migraine, thrombosis, and parkinsonism (Flieger, Wurst, & Shelby, 1997).
Relating to Alternaria toxins, toxicity data are rather sparsely available; to date some of these mycotoxins have been shown to be teratogenic in vivo (EFSA, 2011b). Moreover, genotoxic effects of alternariol (AOH) and alternariol monomethyl ether (AME) in vitro have been described (EFSA, 2011b). In addition, these toxins have been associated with human esophageal cancer in the context of contaminated grain in Linxian, China (Ostry, 2008).
In the EU, regulatory levels for ergot alkaloids and Alternaria toxins in food have not yet been defined. Recently, a maximum level of 0.5 g/kg ergot sclerotia in certain unprocessed cereals has been set by Commission Regulation (EU) 2015/1940 of October 2015 (EC, 2015). Although this regulation does not directly refer to the ergot alkaloid content, it characterizes ergot as a topic of concern, which should forward the provision of maximum levels for ergot alkaloids in the near future. In 2012, the European Commission has recommended that member states should collect data on the predominantly occurring ergot alkaloids in food and feed, namely ergometrine, ergotamine, ergosine, ergocristine, ergocryptine, ergocornine, and their related -inines (EC, 2012). In previous studies, both ergot alkaloids and Alternaria toxins have been detected in grains and grain-based foods intended for human consumption (Crews et al., 2009, Müller and Korn, 2013, Scott et al., 2012). In barley, the occurrence of these mycotoxins has already been demonstrated (Liesener et al., 2010, Medina et al., 2006). Medina et al. (2006), in particular, identified Alternaria as the dominant fungal species isolated from malting barley samples. Still, relatively few information is available about the occurrence of these mycotoxins in beer. Recently, Prelle, Spadaro, Garibaldi, and Gullino (2013) found AOH to be the most frequent occurring Alternaria mycotoxin in beer. In addition, low average amounts of tenuazonic acid (TEA) in beer had been reported before (Siegel, Merkel, Koch, & Nehls, 2010).
Regarding ergot alkaloids, the fate of these mycotoxins throughout the brewing process was studied by Schwarz, Hill, and Rottinghaus (2007) in the scope of a micro-brewing experiment with barley and wheat samples containing ergot sclerotia. The authors recorded low quantities of ergot alkaloids transmitted into beer.
To the best of our knowledge, a survey of ergot alkaloids and AOH in commercial beer samples from the German market has not yet been published. We therefore performed a preliminary survey of mycotoxins belonging to four different groups, namely DON, ZEA as well as ergot alkaloids and AOH using competitive enzyme immunoassays (EIA) as rapid screening methods.
Section snippets
Sample material
Various brands of bottled beer (n = 44) were purchased from local retail stores in 2014 in the area of Giessen, Germany. Most samples (n = 41) were of German produce, imported beers (n = 3) originated from Ireland (n = 1), Mexico (n = 1), and the Czech Republic (n = 1). The domestic beers were made from wheat or barley, one sample was made from rye. All beer was in bottles, these were stored at room temperature.
Sample preparation
Visibly cloudy beers containing particles were first filtered through a paper
Results and discussion
All four EIAs provided sensitive and robust means of detection of the respective mycotoxin analyzed. Except of DON, no extraction was necessary, low LODs in the ng/L (ZEA, ergometrine equivalents, AOH) or μg/L (DON) range could be achieved in diluted beer samples. The EIA standard curve parameters, the minimum sample dilution factor, and the mean LODs in beer are summarized in Table 2.
Mean recovery rates for spiked beer samples were in the range of 50–104% (Table 3). In most cases (DON,
Acknowledgments
We thank Margit Kessler for her excellent technical assistance.
References (58)
- et al.
Stability of mycotoxins during food processing
International Journal of Food Microbiology
(2007) - et al.
The occurrence of the selected fusarium mycotoxins in Czech malting barley
Food Control
(2014) - et al.
In vivo toxicity studies of fusarium mycotoxins in the last decade: a review
Food and Chemical Toxicology
(2015) - et al.
Deoxynivalenol and zearalenone occurrence in beers analysed by an enzyme-linked immunosorbent assay method
Food Control
(2013) - et al.
Occurrence of Alternaria toxins in food products in the Netherlands
Food Control
(2016) - et al.
Survey of the mycobiota of Spanish malting barley and evaluation of the mycotoxin producing potential of species of Alternaria, Aspergillus and Fusarium
International Journal of Food Microbiology
(2006) - et al.
Alternaria mycotoxins in wheat – a 10 years survey in the northeast of Germany
Food Control
(2013) - et al.
Mycotoxins in cereals and related foodstuffs: a review on occurrence and recent methods of analysis
Trends in Food Science & Technology
(2014) - et al.
Quality and occurrence of deoxynivalenol and fumonisins in craft beer
Food Control
(2015) - et al.
A new method for detection of five alternaria toxins in food matrices based on LC-APCI-MS
Food Chemistry
(2013)
Some major mycotoxins and their mycotoxicoses – an overview
International Journal of Food Microbiology
Quantification of the Alternaria mycotoxin tenuazonic acid in beer
Food Chemistry
Mold and mycotoxin problems encountered during malting and brewing
International Journal of Food Microbiology
Deoxynivalenol and its conjugates in beer: a critical assessment of data obtained by enzyme-linked immunosorbent assay and liquid chromatography coupled to tandem mass spectrometry
Analytica Chimica Acta
Determination of zearalenone and its metabolites α- and β-zearalenol in beer samples by high-performance liquid chromatography-tandem mass spectrometry
Journal of Chromatography B
Widespread occurrence of low levels of alternariol in apple and tomato products, as determined by comparative immunochemical assessment using monoclonal and polyclonal antibodies
Journal of Agricultural and Food Chemistry
Comparison of ochratoxin A and deoxynivalenol in organically and conventionally produced beers sold on the Belgian market
Food Additives & Contaminants
Development of new dispersive liquid-liquid microextraction technique for the identification of zearalenone in beer
Analytical Methods
Potential health hazards due to the occurrence of the mycotoxin tenuazonic acid in infant food
European Food Research and Technology
Occurrence of type A, B and D trichothecenes in barley and barley products from the Bavarian market
Mycotoxin Research
Ergot alkaloids in some rye-based UK cereal products
Food Additives & Contaminants Part B
Deoxynivalenol in food
Mycotoxin Research
Determination of deoxynivalenol in bread and beer
Mycotoxin Research
Masked mycotoxins are efficiently hydrolyzed by human colonic microbiota releasing their aglycones
Chemical Research in Toxicology
Induction of mutagenesis and transformation by the extract of Alternaria alternata isolated from grains in Linxian, China
Carcinogenesis
Commission regulation (EC) no 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs
Official Journal of the European Union
Commission recommendation of 15 March 2012 on the monitoring of the presence of ergot alkaloids in feed and food
Official Journal of the European Union
Commission regulation (EU) 2015/1940 of 28 October 2015 amending regulation (EC) no 1881/2006 as regards maximum levels of ergot sclerotia in certain unprocessed cereals and the provisions on monitoring and reporting
Official Journal of the European Union
Scientific opinion on the risks for public health related to the presence of zearalenone in food
EFSA Journal
Cited by (62)
Co-occurrence of deoxynivalenol and its acetylated derivatives in Chinese beer samples
2023, Journal of Food Composition and AnalysisDevelopments and characteristics of craft beer production processes
2022, Food BioscienceFree and conjugated Alternaria and Fusarium mycotoxins during Pilsner malt production and double-mash brewing
2022, Food ChemistryCitation Excerpt :The concentrations of free mycotoxins and peak area ratios of conjugated to free mycotoxins in barley, malt and beer are summarized in Table 1. As regards malt and beer, while studies carried out about ten years ago reported mainly the presence of DON and its modified forms (D3G, 3-ADON and 15-ADON) (Lancova et al., 2008; Kostelanska et al., 2011; Zachariasova et al., 2012), later studies reported other mycotoxins (Bauer et al., 2016; Pascari et al., 2018), which may be related to changing patterns of mycotoxins (associated with climate change and changing patterns of their producers (Khodaei et al., 2021)). In our study, in malts after both conventional and intensified processes, TEA (451 and 1,889 µg/kg, respectively), ENNB (387 and 612 µg/kg, respectively) and ENNB1 (289 and 375 µg/kg, respectively) were predominant.
Alternaria in malting barley: Characterization and distribution in relation with climatic conditions and barley cultivars
2021, International Journal of Food MicrobiologyPeroxidase as a simultaneous degradation agent of ochratoxin A and zearalenone applied to model solution and beer
2020, Food Research International