Content of Individual Phenolic Acids in Worts and Beers and their Possible Contribution to the Antiradical Activity of Beer

Phenolic acids are widely distributed in foods and raw materials. They are easily absorbed by humans due to their simplicity. Once they enter the blood plasma, they act as antioxidants. Beer can be a rich source of phenolic acids in the diet. The aim of this study was to determine the concentrations of phenolic acids in two experimental worts and beers as well as in nine market beers (using HPLC‐UV). An examination of the total antiradical activities of phenolic acids with in vitro model systems (using ABTS and DPPH free radicals), at the concentrations comparable to those detected in beers, was performed. Only low fractions of the main phenolic acids present in barley malt (ferulic, vanillic and p‐coumaric acid) were detected in the experimental worts. Moreover, the concentrations of phenolic acids significantly decreased until the last steps of beer production. The main beer phenolic acids (vanillic and ferulic acid) exerted a lower share of total antiradical activity against both free radicals (calculated as the sum of the individual activities of all acids detected in beer) than the minor phenolic acids (caffeic, chlorogenic, o‐coumaric, sinapic or syringic acid). The synergies, between individual phenolic acids in pairs, were also studied with in vitro model solutions using free radicals. The total antiradical activity of the compounds studied in pairs, was at the most as high as the sum of the antiradical activity of the individual phenolic acids, but in most cases it was considerably lower (i.e. no synergy was detected).     

Development of low-alcohol isotonic beer by interrupted fermentation

Alcohol-free beer with isotonic properties is getting more popular and its production can be carried out by different production strategies; however, interrupted fermentation is still a challenge. Therefore, the objective of this study was to develop a low-alcohol isotonic beer (<0.5% v/v) by interrupted fermentation. Moreover, the major objective is to compare the developed product to commercial beverages (sports drinks, ‘Pilsen' regular beer, alcohol-free beers and low-alcohol isotonic beer). The beverages were evaluated based on pH, alcohol content (% v/v), total titratable acidity (mEq L⁻¹), osmolality (mOsmol kg⁻¹), bitterness International Bitterness Units, colour European Brewery Convention, total phenolic compounds (mg L⁻¹ gallic acid), reducing and total sugars (%) and Na and K contents (mg L⁻¹). The developed low-alcohol isotonic beer presented characteristics similar to sports drinks, with the advantage of being richer in phenolic compounds and suitable osmolality. Despite salts were added in its formulation, the grades attributed to all beers employed in the sensory evaluation, as well as the purchase intention did not present significant differences.     

Immunochemical determination of gluten in malts and beers.

The gluten content in different varieties of barley and malts, and in different types of beers, was determined by a 'sandwich' enzyme immunoassay (RIDASCREEN Gliadin kit). The gluten levels in barley wheat, rye and spelt malts ranged 18.8-45.0, 44.0-68.0, 41.6 and 21.2 g kg-1, respectively. When various types of beer were compared, the gluten concentration increased as follows: alcohol-free beer (<3.0), lager beers (<3.0-8.7 mg l-1), stouts (9.0-15.2 mg l-1) and wheat beers (10.6-41.2 mg l-1). When 10 Czech lager beers were analysed, using both sandwich and competitive ELISA, the results showed that the latter method provided values several times higher than the former. Gluten balance was carried out during the brewing process, starting from the raw materials and terminating at the final beer. Gluten levels decreased due to precipitation during the mashing process, primary and secondary fermentation and, lastly, as a result of adsorption during beer stabilization. The gluten content in beer is, thus, approximately three orders of magnitude lower than in the raw malt.     

Comparative study of the inhibitory effects of wine polyphenols on the growth of enological lactic acid bacteria

This paper reports a comparative study of the inhibitory potential of 18 phenolic compounds, including hydroxybenzoic acids and their derivatives, hydroxycinnamic acids, phenolic alcohols and other related compounds, stilbenes, flavan-3-ols and flavonols, on different lactic acid bacteria (LAB) strains of the species Oenococcus oeni, Lactobacillus hilgardii and Pediococcus pentosaceus isolated from wine. In general, flavonols and stilbenes showed the greatest inhibitory effects (lowest IC?? values) on the growth of the strains tested (0.160-0.854 for flavonols and 0.307-0.855 g/L for stilbenes). Hydroxycinnamic acids (IC?? > 0.470 g/L) and hydroxybenzoic acids and esters (IC?? >1 g/L) exhibited medium inhibitory effect, and phenolic alcohols (IC?? > 2 g/L) and flavanol-3-ols (negligible effect) showed the lowest effect on the growth of the LAB strains studied. In comparison to the antimicrobial additives used in winemaking, IC?? values of most phenolic compounds were higher than those of potassium metabisulphite for O. oeni strains (e.g., around 4-fold higher for quercetin than for potassium metabisulphite), but lower for L. hilgardii and P. pentosaceus strains (e.g., around 2-fold lower for quercetin). Lysozyme IC?? values were negligible for L. hilgardii and P. pentosaceus, and were higher than those corresponding to most of the phenolic compounds tested for O. oeni strains, indicating that lysozyme was less toxic for LAB than the phenolic compounds in wine. Scanning electron microscopy confirmed damage of the cell membrane integrity as a consequence of the incubation with antimicrobial agents. These results contribute to the understanding of the inhibitory action of wine phenolics on the progress of malolactic fermentation, and also to the development of new alternatives to the use of sulphites in enology.     

Presence and distribution of arsenical species in beers

The total content of arsenic and of its inorganic (As(III) and As(V)) and organic (monomethylarsonic acid, MMAA, and dimethylarsinic acid, DMAA) species were determined in a set of 21 alcoholic and alcohol-free beer samples using the technique of Hydride Generation Atomic Absorption Spectrometry. For total arsenic analysis, beer samples were dried and then microwave digested with nitric acid in polytetrafluoroethylene containers. For the speciation analysis, beers were previously subjected to ion exchange chromatography to elute the mentioned inorganic and organic arsenical species. Both microwave digestion and chromatographic separation methods were validated from certified reference materials and prepared standard solutions, respectively. The results obtained are presented in terms of the distribution and occurrence of arsenical species in the samples. The As levels of the beer samples were in the range of 1.5-12.4mug/l. The influence of the production process for the alcoholfree beers in the speciation of arsenic is discussed. In alcoholic beers MMAA was the most abundant species, and for non-alcoholic beers inorganic As(III) was similar to the organic species. An estimated intake of total As of 0.47mug/person/day and 11.4mug/person/day was obtained for average consumers and for heavy drinkers, respectively.     

Growth and survival of foodborne pathogens in beer

This work aimed to assess the growth and survival of four foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus) in beer. The effects of ethanol, pH, and storage temperature were investigated for the gram-negative pathogens (E. coli O157:H7 and Salmonella Typhimurium), whereas the presence of hops ensured that the gram-positive pathogens (L. monocytogenes and S. aureus) were rapidly inactivated in alcohol-free beer. The pathogens E. coli O157:H7 and Salmonella Typhimurium could not grow in the mid-strength or full-strength beers, although they could survive for more than 30 days in the mid-strength beer when held at 4°C. These pathogens grew rapidly in the alcohol-free beer; however, growth was prevented when the pH of the alcohol-free beer was lowered from the "as received" value of 4.3 to 4.0. Pathogen survival in all beers was prolonged at lowered storage temperatures.     

CONTROL OF FERULIC ACID AND 4-VINYL GUAIACOL IN BREWING

Phenolic acids in beer are important because they can be decarboxylated to phenols, which usually impart off-flavours. An improved high performance liquid chromatographic system was used to monitor phenolic acids and phenols during the brewing process. Ferulic acid was the most significant phenolic acid found in beers prepared from malted barley. Extraction of ferulic acid from malt involved an enzymatic release mechanism with an optimum temperature about 45°C. Mashing-in at 65°C significantly decreased the release of free ferulic acid into the wort. Wort boiling produced 4-vinyl guaiacol by thermal decarboxylation, in amounts (0.3 mg/L) close to its taste threshold, from worts that contained high contents of free ferulic acid (> 6 mg/L). The capacity of yeasts to decarboxylate phenolic acids (Pof⁺ phenotype) was strong in wild strains of Saccharomyces and absent in all lager brewing yeast and most ale brewing yeasts. Some top-fermenting strains, especially those used in wheat beer production, possessed a weak decarboxylating activity (i.e. Pof^δ). During storage of beers there were appreciable temperature-dependent losses of 4-vinyl guaiacol. These results indicated that the production of 4-vinyl guaiacol is amenable to close technological control.     

Impact of Different Hop Compounds on the Overfoaming Volume of Beer Caused by Primary Gushing

When weather conditions favour the growth of moulds on barley, beers brewed from the resulting malts often tend to gush. Certain Fusarium species (e.g., F. graminearum and F. culmorum) may cause this problem. Supersaturated with CO₂, a primary gushing beer contains an overcritical concentration of microbubbles; these are reputed to be stabilised by Fusarium‐derived hydrophobins. Research with varying brewhouse parameters has been performed to investigate the factors of primary gushing. As hops are known to contribute to a wide range of both gushing positive and negative substances in beer, the hopping regime has emerged as an important aspect. This paper examines the impact of different hop varieties on gushing. Hop oils and unsaturated fatty acids are reputed to be gushing‐suppressors. Compounds such as dehydrated humulinic acid can intensify the effect. Hop pellets, with a prevalent range of conductometric values (5–10% α‐acid), commonly employed in breweries to adjust bitterness and aroma were selected. By working with the same “gushing malt”, the spectrum of compounds in the finished beer only differed through the hop product used. The overfoaming volumes of different samples were determined according to MEBAK guidelines. Respective hop oil and fatty acid concentrations (by GC) and iso‐α‐acid contents (by HPLC) were compared and a chronological sequence of the changing percentages of beer loss is shown.     

Phenolic Profiles and Total Antioxidant Capacity of Marketed Beers in Serbia

The aim of this research was to determine the total phenolic and flavonoid contents as well as to measure antioxidant activity of 24 different commercial beers consumed in Serbia. The major phenolic acids (gallic acid, protocatechuic acid, 4-hydroxybenzoic acid, 2,5-dihydroxybenzoic acid, vanillic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, sinapic acid, salicylic), (+)-catechin, and (-)-epicatechin were also determined by high pressure liquid chromatography method using a photodiode array detector. Gallic acid, ferulic acid, and protocatechuic acid are the most abundant phenolic acids in all samples, followed by (+)-catechin. The total phenolic content was determined using the Folin-Ciocalteu assay. The total flavonoids were measured using spectrophotometrics as the aluminum chloride assay. The results showed that the highest total phenolic and flavonoid contents were established in dark and light beer samples. 2,2-Diphenyl-1-picrylhidrazyl radical scavenging activity, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radical cation scavenging activity, and ferric reducing/antioxidant power were used to assess the antioxidant potential of beers. These assays, based on different chemical mechanisms, were selected to take into account the wide variety and range of action of antioxidant compounds present in selected beer samples. All beers showed antioxidant power, but a wide range of antioxidant capacities was observed. Statistical differences between ferric reducing-antioxidant power and the other two antioxidant capacity assays were confirmed. This study will be useful for the appraisal of phenolic profile and antioxidant activities of various beers, and it will also be of interest for people who like drinking this beverage.     

Brewer’s yeast in controlled and uncontrolled fermentations,with a focus on novel,nonconventional, and superior strains

Historically, brewing beer came from uncontrolled spontaneous fermentations; nowadays, spontaneous fermentation is still used in speciality products such as Belgian acid beers (e.g., lambic, gueze, and Rodenbach) or in various traditional beers worldwide. On the other hand, industrial fermentations sometimes require a combination of phenotypic traits that might not be commonly encountered in nature. In some cases, the demand for increased productivity, as well as changing consumer preferences, leads to a great interest towards improvement and/or design new strains or yeast variants. This review addresses the following topics: controlled and uncontrolled fermentations, and advances in the selection of novel, nonconventional, and superior strains for beer.     

Assessment of the aroma profiles of low-alcohol beers using HS-SPME–GC-MS

The volatile profile of low-alcohol and alcohol-free beers was compared with that of alcoholic ones. Qualitative and quantitative differences were analyzed by headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME–GC-MS). Fermentation compounds as esters (isoamyl acetate, ethyl hexanoate, ethyl octanoate), alcohols (1-octanol, decanol, isobutanol, isoamyl alcohol) and fatty acids (hexanoic and octanoic acid) were present in higher quantities in alcoholic beers. The low-alcohol beers were characterized by compounds derived from malt and the roasting process, including pyrazines and furanes, and volatile compounds such as linalool, β-humulene and α-terpineol derived from the essential oil of hops. Benzaldehyde, acetylpyrrole, furfural and 2-furanmethanol were characteristics of free-alcohol beers.     

Simple and Fast Sample Preparation Followed by Gas Chromatography‐Tandem Mass Spectrometry (GC‐MS/MS) for the Analysis of 2‐ and 4‐Methylimidazole in Cola and Dark Beer

We have developed a simple and fast sample preparation technique in combination with a gas chromatography‐tandem mass spectrometry (GC‐MS/MS) for the quantification of 2‐methylimidazole (2‐MeI) and 4‐methylimidazole (4‐MeI) in colas and dark beers. Conventional sample preparation technique for GC‐MS requires laborious and time‐consuming steps consisting of sample concentration, pH adjustment, ion pair extraction, centrifugation, back‐extraction, centrifugation, derivatization, and extraction. Our sample preparation technique consists of only 2 steps (in situ derivation and extraction) which requires less than 3 min. This method provided high linearity, low limit of detection and limit of quantification, high recovery, and high intra‐ and interday repeatability. It was found that internal standard method with diluted stable isotope (4‐MeI‐d₆) and 2‐ethylimidazole (2‐EI) could not correctly compensate the matrix effects. Thus, standard addition technique was used for the quantification of 2‐ and 4‐MeI. The established method was successfully applied to colas and dark beers for the determination of 2‐MeI and 4‐MeI. The 4‐MeI contents in colas and dark beers ranged from 8 to 319 μg/L and from trace to 417 μg/L, respectively. Small quantity (0 to 8 μg/L) of 2‐MeI was found only in dark beers. The contents of 4‐MeI (22 μg/L) in colas obtained from fast food restaurants were significantly lower than those (177 μg/L) in canned or bottled colas.     

Rich nutrition from the poorest – Cereal fermentations in Africa and Asia

Cereal fermentations in Africa and Asia involve mainly the processing of maize, rice, sorghum and the millets. Lactic acid bacteria (Lactobacillus, Pediococcus), Enterobacter spp., yeasts (Candida, Debaryomyces, Endomycopsis, Hansenula, Pichia, Saccharomyces and Trichosporon spp.) and filamentous fungi (Amylomyces, Aspergillus, Mucor, and Rhizopus spp.) contribute to desirable modifications of taste, flavour, acidity, digestibility, and texture in non-alcoholic beverages (e.g., uji, and ben-saalga), porridges (e.g., mawè) and cooked gels (e.g., kenkey, idli, and mifen). In addition, alcoholic beverages (beers such as tchoukoutou and jnard; and spirits e.g. jiu) are obtained using malt, or using amylolytic mixed microbial starter cultures as generators of fermentable substrates. Wet processing, marketing of multi-purpose intermediate products, co-fermentation for texture and nutrition, and mixed culture fermentations as practiced in indigenous fermentation processes are of interest for industrial innovation and for better control of natural mixed culture fermentation systems. On the other hand, the nutritional properties of traditional cereal fermented products can be enhanced by increasing their nutrient and energy density, as well as by increasing their mineral status by combining mineral fortification and dephytinization.     

ORGANIC ACIDS AND GLYCEROL IN BEER

The concentrations of pyruvic acid, citric acid, L-malic acid, acetic acid, L-lactic acid and glycerol were determined in beers of different type and origin. Large differences in organic acid and glycerol contents were found, especially between different types of beer. The amounts of acetic acid, lactic acid and glycerol can be used as process parameters. During malting a decrease of the malic acid-citric acid ratio occurred due to the pathway of the TCA-cycle. The acetic acid content and glycerol contents showed increases during fermentation. The latter parameter is useful to indicate if alcohol-free beer has been produced by fermentation. Determination of apparent final attenuation degrees of worts according to the usual laboratory practice resulted in higher glycerol contents than was found for the corresponding beers, making the value of this assay questionable.     

Total hordatine content in different types of beers

Hordatines are phenolic secondary metabolites typical of barley. Hordatines withstand at least moderate processing, and thus they are also found in barley malts and beer. So far, no published data on the hordatine content has been available in beers or different styles of beer. The aim of this study was to produce information on the total hordatine content in beers and statistically compare the hordatine content of different beer types. In the current study, hordatines were analysed in 208 beers by high‐performance liquid chromatography equipped with a diode array detector (HPLC‐DAD). The average total hordatine content of all beer samples was 5.6 ± 3.1 mg L^?1 as p‐coumaric acid equivalents (PCAE), with a minimum values 0 to a maximum value 18.7 mg L^?1 PCAE. The total hordatine content correlated positively to the alcohol content in lagers, ales, stouts and porters, but not in wheat beers. There was no statistically significant difference in hordatine content in different types of beer, excluding the non‐alcoholic group of beers. It is noteworthy that non‐alcoholic beers also contained hordatines. More research would be needed to understand how parameters, such as mashing, should be chosen in order to achieve maximum recovery of hordatines in wort and beer.     

Isolation and Identification of Phenolic Antioxidants in Black Rice Bran

Black rice bran contains phenolic compounds of a high antioxidant activity. In this study, the 40% acetone extract of black rice bran was sequentially fractionated to obtain 5 fractions. Out of the 5 fractions, ethyl acetate fraction was subfractionated using the Sephadex LH‐20 chromatography. The antioxidant activity of phenolic compounds in the extracts was investigated by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical assay, 2,2‐azino‐bis‐(3‐ethylenebenzothiozoline‐6‐sulfonic acid) (ABTS) radical cation assay, reducing power. The subfraction 2 from ethyl acetate fraction had the highest total phenolic contents (TPC) (816.0 μg/mg) and the lowest EC₅₀ values (47.8 μg/mL for DPPH radical assay, 112.8 μg/mL for ABTS radical cation assay, and 49.2 μg/mL for reducing power). These results were 3.1, 1.3, and 2.6 times lower than those of butylated hydroxytoluene (BHT), respectively. At a concentration of 100 μg/mL, the antioxidant activity and TPC of various extracts was closely correlated, with correlation coefficients (R²) higher than 0.86. The major phenolic acid in subfraction 2 was identified as ferulic acid (178.3 μg/mg) by HPLC and LC‐ESI/MS/MS analyses. Our finding identified ferulic acid as a major phenolic compound in black rice bran, and supports the potential use of black rice bran as a natural source of antioxidant.     

Alcohol-free and low-alcohol beers: Aroma chemistry and sensory characteristics

Alcohol-free beers have gained popularity in the last few decades because they provide a healthier alternative to alcoholic beers and can be more widely consumed. Consumers are becoming more aware of the benefits of reducing their alcohol consumption, and this has increased the sales of nonalcoholic alternatives. However, there are still many challenges for the brewing industry to produce an alcohol-free beer that resembles the pleasant fruity flavor and overall sensory experience of regular beers. The aim of this review is to give a comprehensive overview of alcohol-free beer focusing on aroma chemistry. The formation of the most important aroma compounds, such as Strecker aldehydes, higher alcohols, and esters, is reviewed, aiming to outline the gaps in current knowledge. The role of ethanol as a direct and indirect flavor-active compound is examined separately. In parallel, the influence of the most common methods to reduce alcohol content, such as physical (dealcoholization) or biological, on the organoleptic characteristics and consumer perception of the final product, is discussed.     

Phenolic compounds’ characterization of Artemisia rutifolia spreng from Pakistani flora and their relationships with antioxidant and antimicrobial attributes

Artemisia rutifolia (Asteraceae) had been used in traditional medicines for the treatment of different ailments. In the current study, an effort was made to explore the phenolic composition, antioxidant, and antimicrobial activities of different solvent extracts obtained from A. rutifolia leaves. The reverse-phase high-performance liquid chromatographic (RP-HPLC) analysis revealed the higher extent of polyphenolic compounds (i.e., gallic acid, caffeic acid, chlorogenic acid, syringic acid, sinapic acid, p-coumaric acid, m-coumaric acid, ferulic acid, vanillic acid, myricetin, and quercetin) in methanol extract. Methanol extract consistently showed the highest total phenolic contents (98 ± 2 µg GAE/mg of plant extract), total flavonoid contents (28 ± 0.0 µg QE/mg of plant extract), antimicrobial activity, free radical (DPPH) scavenging (IC₅₀ = 39 µg/mL) activity, and reducing power (18.3 ± 0.2 mg GAE/g of plant extract) followed by those of chloroform and hexane extracts, respectively. The current study concluded that extracts of A. rutifolia are novel natural source of antioxidative and antimicrobial agents for the treatment of oxidative stress-related disorders and microbial infections.     

The influence of training and expertise on the multisensory perception of beer: A review

This review critically evaluates the literature documenting the impact of training on people’s perception of beer. In certain circumstances, training has been shown to improve people’s ability to match and identify beers, and to discriminate between beers and between the distinctive attributes of beer. However, a reasonably consistent finding is that the benefits of beer training do not seem to generalise to novel beers (i.e., those not experienced during training). As such, training would appear to improve the capacity to label perception and/or people’s recognition memory for beers, rather than necessarily influencing perception itself. Given how much beer is consumed annually, it is surprising that there has not been more published research into the role that training plays in this particular beverage category. Part of the reason for this may relate to (a) the limited role of experts in this field (e.g., primarily restricted to product testing and quality control; as compared to their much more prominent role in the world of wine), (b) the fact that training is an expensive and time-consuming process, and (c) it is wrongly assumed that the effects of training are similar across different beverage categories (e.g., beer and wine). We suggest that further efforts are therefore required before it will be possible to confidently conclude that training lowers the perceptual thresholds, and enhances the perceptual discrimination abilities, of beer experts above those of novices.     

PHENOLIC ACIDS IN BEERS AND WORTS

The total contents of phenolic acids measured by high-performance liquid-chromatography were 5–8 mg/litre in beers brewed in Ireland whereas 16–40 mg/litre were present in four other beers. In all beers the predominant phenolic acids were vanillic, p-coumaric and ferulic acids. Free phenolic acids were extracted from Emma barley grains and malt in very small amounts (15–28 mg/kg) but larger quantities (191 mg/kg) were released on mashing the malt. Little change occurred in the contents of phenolic acids on processing a lager wort through to the finished beer. Treatment with excess Polyclar AT removed astringent flavour and phenolic acids from an experimental ale but this flavour loss could not be accounted for by the adsorption of phenolic acids. The flavour threshold for a nine-component phenolic acid mixture in lager was between 50 mg/litre and 100 mg/litre.