Mycotoxin analysis of industrial beers from Brazil: The influence of fumonisin B1 and deoxynivalenol in beer quality

doi:10.1016/j.foodchem.2016.09.062

Food Chemistry

Volume 218, 1 March 2017, Pages 64-69

https://doi.org/10.1016/j.foodchem.2016.09.062 Get rights and content

Highlights

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Deoxynivalenol was not detected in any of the industrial beer samples.
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High levels of fumonisin B₁ were found in 49% of the samples.
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Fumonisin B₁ contamination was possibly due to the use of adjuncts.
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Regulation for mycotoxins in beer is needed.

Abstract

Worldwide, barley is the main source of carbohydrate in the brewing process. However, corn is often used as an adjunct to improve and accelerate the fermentation process. Considering that, these two substrates are susceptible to fungal contamination as well as mycotoxins. The objective of the current study is to determine the incidence of the mycotoxins deoxynivalenol (DON) and fumonisin B₁ (FB₁) in industrial beers.

The method applied for mycotoxin analyses included high performance liquid chromatography_. The mean levels for recovery experiments were 89.6% for DON and 93.3% for FB₁. DON was not detected in any of the analyzed samples whereas FB₁ was found in 49% of the 114 samples. The current survey demonstrated levels of FB₁ contamination in industrial beer, possibly due to the addition of contaminated adjuncts. It is necessary to establish maximum levels of mycotoxins in beer in Brazil and other countries in order to reduce health risks.

Introduction

Beer is a popular alcoholic beverage in the world context for reasons that transcend social and economic aspects. In Brazil, the beer market continues to grow every year and production reached 13.5 billion liters in 2013. In terms of national average consumption, data has shown a large growth potential of 68.3 L per person in 2014 (CervBrasil, 2014).

Worldwide, the source of fermentable sugars in industrial beer is starch-rich cereals, mainly malted barley. Nevertheless, adjuncts including corn, rice, unmalted barley, wheat starch, oats and sorghum have been also used by the large-scale brewing industry to provide additional sources of fermentable carbohydrates for the yeast (Figueroa et al., 1995, Poreda et al., 2014).

The brewing industry has many reasons for the application of adjuncts including better availability on the local market, sensory modification of the beer and, the most important, the lower price of this product in Brazil (Dhellot and Kobawila, 2013, Glatthar et al., 2005). It has been proven that the use of 30% of corn adjunct can give an 8% reduction in total production costs, although this number may vary depending on the local prices of raw materials and other costs of production (Baca, 2001). However, application of adjuncts also has some disadvantages. One of the most relevant is the contamination by fungi and the production of mycotoxins. These will have a negative impact on the final product, mainly because of the health problems associated with mycotoxin contamination (Lancova et al., 2008). Several studies revealed that barley can be contaminated by fungi (Piacentini, Savi, Olivo, & Scussel, 2015), as well as the adjuncts used for brewing (Kawashima et al., 2007, Oliveira et al., 2016, Queiroz et al., 2012, Van der Westhuizen et al., 2003).

The major mycotoxins found in barley are the trichothecenes type B group, primarily DON and its metabolites (Lancova et al., 2008). Trichothecenes are toxins of the sesquiterpenoid metabolism, produced by some species of Fusarium and other fungi in the order Hypocreales (Rocha et al., 2015). DON is a mycotoxin largely produced by the Fusarium graminearum species complex (FGSC) and this group is a devastating pathogen causing Fusarium head blight in wheat and barley. DON is considered a potent inhibitor of protein biosynthesis, exhibiting acute adverse effects in animals (Rubella, Goswami, & Kistler, 2004).

Considering that corn is one of the adjuncts used for brewing, detection of fumonisins B₁ and B₂ (FB₁ and FB₂) are expected in beer, as these are the most prevalent mycotoxins in this cereal and its processed products (Matumba et al., 2014, Oliveira et al., 2016). Fumonisins is an important mycotoxin group produced mainly by the Fusarium fujikuroi species complex (FFSC) (Rocha et al., 2016). Fusarium verticillioides, the major fumonisin-producing species within the FFSC, is able to produce levels above 5000 μg/kg as observed by many authors worldwide (Hinojo et al., 2006, Vismer et al., 2004).

FB₁ is known to cause toxicity in animals due to the inhibition of sphingolipid metabolism and cell cycle regulation, resulting in several adverse effects such as leukoencephalomalacia in horses and pulmonary oedema in swine (Desjardins, 2006). It is also associated with oesophageal cancer and neural tube birth defects in humans (Marasas et al., 2004).

Nowadays, there is a lack of regulation for toxin levels in beer, with maximum levels set only for raw materials (Brasil, 2011, Brasil, 2013, Commission of the European Communities, 2006). However, the Scientific Committee for Food (SCF) and FAO/WHO Joint Expert Committee on Food Additives (JECFA) (Bolger et al., 2001) indicated a tolerable daily intake (TDI) of 1 μg/kg⁻¹ and 2 μg/kg⁻¹ bw for DON and FB₁, respectively.

For the reasons stated above and considering that the brewing industry in Brazil is continuing to rise, the objective of the study was to determine the influence of the mycotoxins DON and FB₁ on industrial beer quality from Brazil.

Section snippets

Beer samples

A total of 114 industrial beer cans and bottles (Lager type) consisting of 20 different brands and different batches were randomly chosen to represent the main brewing industries of Brazil. These samples were acquired from markets between September 2015 and January 2016. None of the beers had surpassed their expiry date. Until the sample preparation, they were stored in the dark at room temperature.

Chemicals and reagents

DON and FB₁ standards were obtained from Sigma Aldrich Chemicals (St. Louis, MO, USA). The DON

Validation methods

The samples used for validation were analyzed beforehand to ensure that they did not contain any of the studied compounds. Afterwards, blank samples were selected for spiking and recovery purposes. The spiking concentrations and their recoveries are shown in Table 1 and were carried out in triplicate. The linearity was established by the calibration curve with injections of different standard concentrations aforementioned (0.025–6 μg/ml for FB₁ and 0.1–20 μg/ml for DON). Each standard

Discussion

The current study demonstrated that industrial Brazilian beer was not contaminated by DON, whereas FB₁ levels were relevant. Indeed, previous studies have shown that the transmission of mycotoxins into the final beer will depend on crop infection, agricultural practices and the technological conditions applied during the brewing process (Kostelanska et al., 2009, Lancova et al., 2008, Omurtag and Beyoglu, 2007). On the other hand, it has been established that during the brewing process, DON can

Conclusion

The current survey demonstrated relevant levels of FB₁ contamination in industrial beer, possibly due to the addition of contaminated adjuncts. In order to replace barley or enhance the fermentation process, corn is often used in Brazilian beer industries because of its wide distribution and low costs in the country. This explains the incidence of FB₁ in the analyzed samples. The results indicate the need for strategies to reduce mycotoxin contamination in the raw material used for beer

Financial support

The authors thank FAPESP for financial support.

References (47)

J.I. Bauer et al.
Investigations on the occurrence of mycotoxins in beer
Food Control
(2016)
T. Bertuzzi et al.
Mycotoxin occurrence in beer produced in several European countries
Food Control
(2011)
M. Bryła et al.
Fumonisins and their masked forms in maize products
Food Control
(2016)
M.J. Hinojo et al.
Fumonisin production in rice cultures of Fusarium verticillioides under different incubation conditions using an optimized analytical method
Food Microbiology
(2006)
F.E. Lanza et al.
Prevalence of fumonisin-producing Fusarium species in Brazilian corn grains
Crop Protection
(2014)
W.F.O. Marasas et al.
Fumonisins disrupt sphingolipid metabolism, folato transport, and neural tube development in embryo culture and in vivo: a potential risk factor for human neural tube defects among populations consuming fumonisin-contaminated maize
Journal of Nutrition
(2004)
L. Matumba et al.
Limited survey of mycotoxins in traditional maize based opaque beers in Malawi
Food Control
(2014)
K. O’Donnell et al.
Phylogenetic analyses of RPB1 and RPB2 support middle Cretaceous origin for a clade comprising all agriculturally and medically important fusaria
Fungal Genetics and Biology
(2013)
G.Z. Omurtag et al.
Occurrence of deoxynivalenol (vomitoxin) in beer in Turkey detected by HPLC
Food Control
(2007)
K.C. Piacentini et al.
Quality and occurrence of deoxynivalenol and fumonisins in craft beer
Food Control
(2015)

V.A.V. Queiroz et al.

Occurrence of fumonisins and zearalenone in maize stored in family farm in Minas Gerais, Brazil

Food Control

(2012)

Y. Rodríguez-Carrasco et al.

Occurrence of Fusarium mycotoxins and their dietary intake through beer consumption by the European population

Food Chemistry

(2015)

J.M. Soriano et al.

Mechanism of action of splingolipids and their metabolites in the toxicity of fumonisin B1

Progress in Lipid Research

(2005)

M.R. Torres et al.

Occurrence of fumonisins in Spanish beers analyzed by an enzyme-linked immunosorbent assay method

International Journal of Food Microbiology

(1998)

J.F. Alberts et al.

Effects of temperature and incubation period on production of fumonisin B1 by Fusarium moniliforme

Applied and Environment Microbiology

(1990)

AOAC

Official Methods of Analysis of AOAC International

E. Baca

The role of unmalted adjuncts in the creation of sensory characteristics of beer

Bolger, M., Coker, R., DiNovi, M., Gaylor, D., Gelderblom, W., & Olsen, M., et al. (2001). Fumonisins: safety...

Brasil. (2011). Agência Nacional de Vigilância Sanitária. Resoluçao RDC no. 7, de 18 de fevereiro de 2011. Diário...

Brasil. (2013). Agência Nacional de Vigilância Sanitária e ANVISA. Prorrogação para 1 de janeiro de 2017 o prazo para...

Brasil. Decreto n. 2314, de 04 de setembro de 1997. (1997). Regulamenta a Lei n° 8.918, de 14 de julho de 1994, que...

CervBrasil – Associação brasileira da indústria da cerveja (2014). Anuário (2014). p03-33. Disponível em:...

Commission of the European Communities

Commission regulation no 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs

Official Journal of the European Union

(2006)

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Mycotoxin analysis of industrial beers from Brazil: The influence of fumonisin B1 and deoxynivalenol in beer quality

Highlights

Abstract

Introduction

Section snippets

Beer samples

Chemicals and reagents

Validation methods

Discussion

Conclusion

Financial support

Food Control

Food Control

Food Control

Food Microbiology

Crop Protection

Journal of Nutrition

Food Control

Fungal Genetics and Biology

Food Control

Food Control

Food Control

Food Chemistry

Progress in Lipid Research

International Journal of Food Microbiology

Effects of temperature and incubation period on production of fumonisin B1 by Fusarium moniliforme

Applied and Environment Microbiology

Official Methods of Analysis of AOAC International

The role of unmalted adjuncts in the creation of sensory characteristics of beer

Commission regulation no 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs

Official Journal of the European Union

Mycotoxin analysis of industrial beers from Brazil: The influence of fumonisin B₁ and deoxynivalenol in beer quality