Measuring Hordein (Gluten) in Beer – A Comparison of ELISA and Mass Spectrometry

Background

Subjects suffering from coeliac disease, gluten allergy/intolerance must adopt a lifelong avoidance of gluten. Beer contains trace levels of hordeins (gluten) which are too high to be safely consumed by most coeliacs. Accurate measurement of trace hordeins by ELISA is problematic.

Methods

We have compared hordein levels in sixty beers, by sandwich ELISA, with the level determined using multiple reaction monitoring mass spectrometry (MRM-MS).

Results

Hordein levels measured by ELISA varied by four orders of magnitude, from zero (for known gluten-free beers) to 47,000 µg/mL (ppm; for a wheat-based beer). Half the commercial gluten-free beers were free of hordein by MS and ELISA. Two gluten-free and two low-gluten beers had zero ELISA readings, but contained significant hordein levels (p<0.05), or near average (60–140%) hordein levels, by MS, respectively. Six beers gave false negatives, with zero ELISA readings but near average hordein content by MS. Approximately 20% of commercial beers had ELISA readings less than 1 ppm, but a near average hordein content by MS. Several barley beers also contained undeclared wheat proteins.

Conclusions

ELISA results did not correlate with the relative content of hordein peptides determined by MS, with all barley based beers containing hordein. We suggest that mass spectrometry is more reliable than ELISA, as ELISA enumerates only the concentration of particular amino-acid epitopes; this may vary between different hordeins and may not be related to the absolute hordein concentration. MS quantification is undertaken using peptides that are specific and unique, enabling the quantification of individual hordein isoforms. This outlines the problem of relying solely on ELISA determination of gluten in beverages such as beer and highlights the need for the development of new sensitive and selective quantitative assay such as MS.     

Application of chitooligosaccharides as antioxidants in beer to improve the flavour stability by protecting against beer staling during storage

Objective

To improve beer flavour stability by adding chitooligosaccharides that prevent formation of staling compounds and also scavenge radicals in stale beer.

Results

Chitooligosaccharides, at 0.001–0.01%, inhibited the formation of staling compounds in forced aged beer. The formation of 5-hydroxymethylfurfural, trans-2-nonenal and phenylacetaldehyde were decreased by 105, 360 and 27%, respectively, when compared with those in stale beer without chitooligosaccharide addition. The capability of chitooligosaccharides to prevent staling compound formation depended on their molecular size (2 or 3 kDa). The DPPH/hydroxyl radical scavenging activity in fresh beer significantly lower than that in forced aged beer in the presence of chitooligosaccharides. When compared with stale beer without added chitooligosaccharides, the radical scavenging activity could be increased by adding chitooligosaccharides to forced aged beer.

Conclusions

Chitooligosaccharides play an active part in the prevention of beer flavour deterioration by inhibiting the formation of staling compounds and increasing radical scavenging activity.

     

Beer as a Rich Source of Fluoride Delivered into the Body

Fluoride is an element which in the minimum amount is necessary for the proper construction of the teeth and bones. But on the other hand, it increases the synthesis of reactive oxygen species, inflammatory mediators, and impairs the action of enzymes. Beer is the most popular alcoholic beverage in the world. Due to its prevalence and volume of consumption, it should be considered as a potential source of F- and taken into account in designing a balanced diet. Therefore, the aim of this study was to analyze beer samples in terms of F- levels. The concentrations of fluoride were examined using ion-selective electrode Thermo Scientific Orion and statistical analysis was based on two-way ANOVA and t test. When compared to imported beers, Polish beers were characterized by the lowest mean F- concentration (0.089 ppm). The highest mean F- concentrations were recorded in beers from Thailand (0.260 ppm), Italy (0.238 ppm), Mexico (0.210 ppm), and China (0.203 ppm). Our study shows that beer is a significant source of fluoride for humans, which is mainly associated with the quality of the water used in beer production.     

The use of chitooligosaccharide in beer brewing for protection against beer-spoilage bacteria and its influence on beer performance

Objectives

To identify a biological preservative that can protect beer from microbial contamination, which often results in the production of turbidity and off-flavor.

Results

The antimicrobial activity of a chitooligosaccharide against beer-spoilage bacteria and its effect on the fermentation performance of brewer’s yeast was studied. Chitooligosaccharide with an average 2 kDa molecular weight was the best at inhibiting all tested beer-spoilage bacteria. The application of chitooligosaccharide in the brewing process did not influence the fermentation of brewer’s yeast. The change in beer performance induced by the contamination of Lactobacillus brevis could be effectively controlled by application of chitooligosaccharide in the beer brewing process.

Conclusion

The experimental data suggested that chitooligosaccharide should be an excellent preservative to inhibit beer-spoilage bacteria in the brewing process and in the end product.

     

Bioflavoring and beer refermentation

Various techniques are used to adjust the flavors of foods and beverages to new market demands. Although synthetic flavoring chemicals are still widely used, flavors produced by biological methods (bioflavors) are now more and more requested by consumers, increasingly concerned with health and environmental problems caused by synthetic chemicals. Bioflavors can be extracted from plants or produced with plant cell cultures, microorganisms or isolated enzymes. This Mini-Review paper gives an overview of different systems for the microbial production of natural flavors, either de novo, or starting with selected flavor precursor molecules. Emphasis is put on the bioflavoring of beer and the possibilities offered by beer refermentation processes. The use of flavor precursors in combination with non-conventional or genetically modified yeasts for the production of new products is discussed.     

A short history of beer brewing: Alcoholic fermentation and yeast technology over time

The history of beer: from a staple food to a consumer product with an enormous variety of styles and tastes. Subject Categories: Biotechnology & Synthetic Biology, History & Philosophy of Science

As far back as we can retrace history, beer has always been an important part of human life: it was and still is a valuable food staple that has been constantly improved and adapted to human needs. For most of the time, intoxication was not the main purpose and could only be achieved to a limited extent, if at all, given that beer had a low alcohol content for most of human history. Instead, beer, owing to its specific ingredients and characteristics—alcohol, carbon dioxide and a low pH value—was often the only safe liquid to drink when clean water was rare.

For most of the time, intoxication was not the main purpose and could only be achieved to a limited extent, if at all, given that beer had a low alcohol content for most of human history.

In addition, beer and other fermented foods are an important source of essential vitamins, such as vitamin B or riboflavin, trace elements and other health‐promoting ingredients. Especially for poorer people who mainly lived on bread or porridge, supplementing their diet with beer was beneficial to their health. Beer was also an important staple for certain professions, such as seafarers, who had to live of vitamin‐poor foodstuffs for longer times. Not surprisingly, many seafaring nations contributed to the spread and improvement of beer brewing (Fig 1).Open in a separate windowFigure 1Beer brewing over timeThe most important discoveries and developments during a history of 10,000 years of brewing beer.     

Farm-scale production of fuel ethanol and wet grain from corn in a batch process

The batch production of fuel grade ethanol and distillers' wet grain (wet solids) in a farm-scale process (1240-15,580 L/batch) is described. The employs yeast fermentation of amylase-treated corn mash and a two-stage distillation. Primary emphasis in this study was on the cooking, fermentation, and centrifugation steps. Without recycling, fermentation of the mash yield beers with 10.0-10.5% ethanol. Recycling of stillage supernatant at full, 75, or 50% strengths produced enriched mashes that after 48-h fermentation yielded beers with 5-;14% more ethanol. Recycling twice with full-strength supernatant at pH 7.0 increased the ethanol yield in the final beer 16.5%; however, the time to complete the final fermentation was extended form 48 to 72 h and salt buildup occurred. By recycling at pH 5.4, it was possible to avoid rapids salt buildup and obtain beers with 10.3-10.5% ethanol. Recycling resulted in increased levels of glucose, starch, crude protein, and fat in the beer and a reduced moisture content while the wet solids showed an increased starch content. Centrifugation after cooking or fermentation yield in the subsequently produced beer. Fermentation of a volume-resorted mash supernatant gave a beer with only 9.25% ethanol. Mash wet solids varied somewhat chemically from beer and stillage solids. An economic and energy balance analysis of various modes of plant operation are provided and plant considerations are suggested.     

Life cycle environmental impacts and costs of beer production and consumption in the UK

Purpose

Global beer consumption is growing steadily and has recently reached 187.37 billion litres per year. The UK ranked 8th in the world, with 4.5 billion litres of beer produced annually. This paper considers life cycle environmental impacts and costs of beer production and consumption in the UK which are currently unknown. The analysis is carried out for two functional units: (i) production and consumption of 1 l of beer at home and (ii) annual production and consumption of beer in the UK. The system boundary is from cradle to grave.

Methods

Life cycle impacts have been estimated following the guidelines in ISO 14040/44; the methodology for life cycle costing is congruent with the LCA approach. Primary data have been obtained from a beer manufacturer; secondary data are sourced from the CCaLC, Ecoinvent and GaBi databases. GaBi 4.3 has been used for LCA modelling and the environmental impacts have been estimated according to the CML 2001 method.

Results and discussion

Depending on the type of packaging (glass bottles, aluminium and steel cans), 1 l of beer requires for example 10.3–17.5 MJ of primary energy and 41.2–41.8 l of water, emits 510–842 g of CO₂ eq. and has the life cycle costs of 12.72–14.37 pence. Extrapolating the results to the annual consumption of beer in the UK translates to a primary energy demand of over 49,600 TJ (0.56 % of UK primary energy consumption), water consumption of 1.85 bn hl (5.3 % of UK demand), emissions of 2.16 mt CO₂ eq. (0.85 % of UK emissions) and the life cycle costs of £553 million (3.2 % of UK beer market value). Production of raw materials is the main hotspot, contributing from 47 to 63 % to the impacts and 67 % to the life cycle costs. The packaging adds 19 to 46 % to the impacts and 13 % to the costs.

Conclusions

Beer in steel cans has the lowest impacts for five out of 12 impact categories considered: primary energy demand, depletion of abiotic resources, acidification, marine and freshwater toxicity. Bottled beer is the worst option for nine impact categories, including global warming and primary energy demand, but it has the lowest human toxicity potential. Beer in aluminium cans is the best option for ozone layer depletion and photochemical smog but has the highest human and marine toxicity potentials.

     

Environmental evaluation of aluminium cans for beverages in the German context

Background

In the years 2000 and 2002, the German Environment Agency in Berlin (UBA) published the results of a comprehensive LCA study on beverage containers comprising aluminium cans with volumes of 330 ml and 500 ml. Starting with the aluminium can scenarios and the respective results obtained during the UBA study, additional analyses were performed by IFEU in 2003, a German consultant having been a member of the project team working on the UBA study. The objective was to examine the influence of selected parameters on the LCA profile of carbonated soft drink containers. Data and method were in complete analogy with the LCI and LCA part of the UBA study.

Materials

In 2006, the aluminium industry commissioned a study on further influential factors that help determine the sale of certain types of beer, studying the effects of two selected parameter settings on the comparative results of the aluminium can against the refillable glass bottle. In this scenario, special attention was given to two influential factors, the distribution distance—distinguished by regional and nationwide distribution—and trippage rate.

Results and discussion

The results of the initial LCA from the years 2000 and 2002 showed, for the examined parameters container weight, rate of post-consumer recovery of used containers, degree of recycled content and quality of the recycling routes, that each had a considerable influence on the environmental impact profile of the aluminium can within the given framework. Can weight and recycling rate were sensitive factors in the impact categories of climate change, fossil resources, summer smog (POCP), acidification and terrestrial eutrophication. Can volume affected virtually all impact categories examined.

Conclusions

By now, individual improvement options have already been put into practice in Germany. The environmental profile of the average 330 ml aluminium can on the German market can be expected to be ahead of that of the aluminium can at the time of the UBA study. The introduction of a 500-ml can on the market denotes a fundamental step forward in improving LCA results of the aluminium can as a container for beverages.

     

Perceptions on LCA implementation: evidence from a survey on adopters and nonadopters in Italy

Purpose

This paper introduces the results of an Italian survey on the implementation of life cycle assessment (LCA). Both LCA adopters and nonadopters were involved, in order to understand the main benefits and barriers to the adoption of LCA and how the experiences of LCA adopters differ from the expectations of nonadopters.

Methods

Approximately 2000 Italian companies were contacted and 122 companies participated in the online survey, which represent the 6.5 % of the statistical population of our study. To define the statistical population, firms that had implemented an LCA or an environmental initiative according to an official international standard were only considered. Statistically significant differences in answers between LCA adopters and nonadopters were tested by performing the Mann–Whitney test.

Results and discussion

Companies recognize that LCA can provide useful information to drive strategic decisions and product design, and it is perceived as an opportunity to improve the current monitoring systems. In addition, companies recognize the potential of LCA in marketing, making the communication of green attributes more substantial and robust. Focusing on the barriers experienced by LCA adopters, data collection can be cited. Communication issues also pose a barrier to the further implementation of LCA. The analysis of the results and the comparison of the results for the two groups of respondents highlight that on average, the difficulties are considered as more important than the benefits and that nonadopters tend to overestimate the difficulties and underestimate the benefits connected to the implementation of LCA.

Conclusions

The findings shed light on LCA-related issues both for companies and public servants. The misconception of LCA by nonadopters suggests that an increased awareness is the key to the success of LCA and to its more widespread adoption by companies. It is essential to create and disseminate know-how and sensitize companies to the real barriers and benefits of adopting an LCA. The awareness of potential LCA adopters can be raised by training and education initiatives, as well as by increased possibilities to experiment with these kinds of tools (public programs for financial support, fiscal incentives). On the other hand, market and communication research would contribute to better understand how the environmental impacts of products can be more effectively communicated to clients and consumers.

     

Uncertainty in environmentally conscious decision making: beer or wine?

Purpose

Life cycle assessment (LCA) is being used increasingly in decision support situations. In actual cases, the sources of uncertainty are easily hidden in the complexity. Methods for taking uncertainty into account are recommended by LCA guidelines, but actual application remains rare. The aim of this study is to demonstrate the sources of uncertainty in a practical simple selection case wherein a customer makes a decision between beer and wine in a restaurant, considering the selected criteria and the given information. The uncertainty in LCA results is connected to the broader scope of decision analysis.

Methods

Life cycle inventories were collected for beer and wine production from existing literature. The functional unit was chosen to be one serving of alcohol: beer or wine. For illustrative purposes, only the global warming potential indicator was included in the LCA through carbon footprint (CF). Probabilistic uncertainty analysis was applied to the CF system using Monte Carlo simulation. Water footprint was also roughly considered. In addition, three non-environmental indicators were included in the decision: weight control, price, and taste. The comparison between the two products was constructed as a multiple-criteria decision analytical problem.

Results and discussion

The results indicated that beer had, on average, a higher CF value than wine did. However, the difference was not significant, and within the uncertainty range, also the opposite conclusion was possible. The ratio of wine to beer CF was dominated by the uncertainty in the N₂O emissions of wine production. When all of the decision criteria were included, the level of uncertainty prevented robust overall conclusions about preference for beer or wine. However, depending on the utility differences assigned to subjective indicators, there existed also cases wherein decisions could be made at a 10?% risk level regardless of high overall uncertainty.

Conclusions

In many cases, the uncertainties of LCA are dwarfed by the overall uncertainty of the decision situation. However, as shown by our example, in many cases, reasonable decisions can be made in spite of high uncertainties. The uncertainties of single LCA indicators should be considered in relation to the decision-making problem, which depends on the uncertainty of LCA indicators but also significantly on the weighting of the indicators and the related uncertainty. Successful decision making depends on both the magnitude of uncertainty and the differences in expected utility value between alternatives. More attention should be paid to uncertainty analysis considering the weighting factors.     

EFFECTIVENESS OF PALATE CLEANSERS FOR EVALUATING SOURNESS

The objective of this study was to compare four palate cleansing strategies (crackers, carrots, water and nothing) for their effectiveness in evaluating the sourness of a fruit-flavored beverage. Forty panelists rated the sourness intensity of five orange drink samples that varied in citric acid content. Each subject participated in four sessions, one for each palate cleanser. As expected, sourness increased with increasing citric acid content. There were no significant differences between any of the palate cleansers for the discrimination among the sourness levels. Sourness neither increased (buildup) nor decreased (adaptation) during the session with any of the palate cleansing strategies. Using carrots or water provided similar discrimination to crackers, but the panelists rated the sourness higher after using carrots or water as a palate cleanser than after using the cracker.

PRACTICAL APPLICATIONS

Palate cleansers are widely used during sensory evaluation of foods. We suspect that part of the reason people use them is because they lower the intensity of residual flavors, leading people to believe the flavors have been removed from their mouth. Our results show that residual cleansers can produce changes in the flavor of subsequently tasted products. Because the nature and extent of these changes will logically depend on the specific product and the specific palate cleansing strategy, experiments need to be conducted to provide these measures.     

PREFERENCES FOR SOUTHWESTERN SALTED SNACK FLAVORS: INDIVIDUAL DIFFERENCES AT THE CONCEPTUAL LEVEL AND THE CONSTRUCTION OF A PRODUCT LINE

Individuals differ in what they like, both from a sensory basis, but also from a conceptual basis. For a product to be purchased, the concept (or even the flavor name) must be acceptable just as the physical product itself must be acceptable. This paper presents the results of a study in which the panelists rated degree of interest in 12 different new flavors for a snack chip positioned at the start of the study to have a "taste of the southwest." The results show the substantial variation in acceptance for the different flavors presented as names only (viz., concepts, rather than actual products). Segmentation by attitude of consumers on the basis of the pattern of liking revealed differences among consumers, but did not show the typical opposing patterns found when sensory segmentation is done using actual products having different flavors. These individual differences require that a line of different flavors be developed. There is no simple organizing principle, however, underlying the individual differences. Thus to create the line of flavors requires a different approach. The approach selects various combinations of flavors, identifies the proportion of consumers who would accept at least one flavor of the line (so-called "satisfied consumer"), and then maximizes the number of such "satisfied consumers" for a line comprising a given number of flavors.     

Trigeminal isolated sensory neuropathy (TISN) and FOSMN syndrome: despite a dissimilar disease course do they share common pathophysiological mechanisms?

Background

Patients presenting with bilateral trigeminal hypoesthesia may go on to have trigeminal isolated sensory neuropathy, a benign, purely trigeminal neuropathy, or facial-onset sensory motor neuronopathy (FOSMN), a malignant life-threatening condition. No diagnostic criteria can yet differentiate the two conditions at their onset. Nor is it clear whether the two diseases are distinct entities or share common pathophysiological mechanisms.

Methods

Seeking pathophysiological and diagnostic information to distinguish these two conditions at their onset, in this neurophysiological and morphometric study we neurophysiologically assessed function in myelinated and unmyelinated fibres and histologically examined supraorbital nerve biopsy specimens with optic and electron microscopy in 13 consecutive patients with recent onset trigeminal hypoesthesia and pain.

Results

The disease course distinctly differed in the 13 patients. During a mean 10 year follow-up whereas in eight patients the disease remained relatively stable, in the other five it progressed to possibly life-threatening motor disturbances and extra-trigeminal spread. From two to six years elapsed between the first sensory symptoms and the onset of motor disorders. In patients with trigeminal isolated sensory neuropathy (TISN) and in those with FOSMN neurophysiological and histological examination documented a neuronopathy manifesting with trigeminal nerve damage selectively affecting myelinated fibres, but sparing the Ia-fibre-mediated proprioceptive reflex.

Conclusions

Although no clinical diagnostic criteria can distinguish the two conditions at onset, neurophysiological and nerve-biopsy findings specify that in both disorders trigeminal nerve damage manifests as a dissociated neuronopathy affecting myelinated and sparing unmyelinated fibres, thus suggesting similar pathophysiological mechanisms.

     

Modeling biological systems with uncertain kinetic data using fuzzy continuous Petri nets

Background

Uncertainties exist in many biological systems, which can be classified as random uncertainties and fuzzy uncertainties. The former can usually be dealt with using stochastic methods, while the latter have to be handled with such approaches as fuzzy methods.

Results

In this paper, we focus on a special type of biological systems that can be described using ordinary differential equations or continuous Petri nets (CPNs), but some kinetic parameters are missing or inaccurate. For this, we propose a class of fuzzy continuous Petri nets (FCPNs) by combining CPNs and fuzzy logics. We also present and implement a simulation algorithm for FCPNs, and illustrate our method with the heat shock response system.

Conclusions

This approach can be used to model biological systems where some kinetic parameters are not available or their values vary due to some environmental factors.

     

Population based cohort study of the association between alcohol intake and cancer of the upper digestive tract

Objective: To examine the relation between different types of alcoholic drinks and upper digestive tract cancers (oropharyngeal and oesophageal). Design: Population based study with baseline assessment of intake of beer, wine, and spirits, smoking habits, educational level, and 2-19 years’ follow up on risk of upper digestive tract cancer. Setting: Denmark. Subjects: 15 117 men and 13 063 women aged 20 to 98 years. Main outcome measure: Number and time of identification of incident upper digestive tract cancer during follow up. Results: During a mean follow up of 13.5 years, 156 subjects developed upper digestive tract cancer. Compared with non-drinkers (drinkers of <1 drink/week), subjects who drank 7-21 beers or spirits a week but no wine were at a risk of 3.0 (95% confidence interval 1.5 to 6.1), whereas those who had the same total alcohol intake but with wine as ⩾30% of their intake had a risk of 0.5 (0.2 to 1.4). Drinkers of >21 beers and spirits but no wine had a relative risk of 5.2 (2.7 to 10.2) compared with non-drinkers, whereas those who drank the same amount, but included wine in their alcohol intake, had a relative risk of 1.7 (0.6 to 4.4). Conclusion: A moderate intake of wine probably does not increase the risk of upper digestive tract cancer, whereas a moderate intake of beer or spirits increases the risk considerably.

Key messages

• Alcohol is a strong risk factor for oropharyngeal and oesophageal cancer
• The carcinogenic effect of alcohol has been assumed to be independent of type of alcohol drunk
• Resveratrol, a substance in grapes and wine, has been shown to inhibit the initiation, promotion, and progression of cancer
• Wine drinkers may be at a lower risk of developing upper digestive tract cancer than drinkers who have a similar intake of beer or spirits