Contribution of monophenols to beer flavour based on flavour thresholds, interactions and recombination experiments.

Although monophenols are known to contribute to the flavour of many foods and beverages, little is known about their influence on beer flavour. Therefore, the contribution of 11 monophenols to the overall beer flavour was studied by determining their flavour thresholds. Large differences in sensitivity were observed between individual tasters. Next, flavour interactions between monophenols were examined in nine binary mixtures, which showed that strong interactions like synergy and antagonism occur. Based on these results, the flavour contribution of the monophenols was estimated by calculating flavour units. These proved to be rather low for most of the studied monophenols. However, recombination experiments demonstrated that monophenols enriched beer flavour with spicy, smokey and vanilla flavour aspects. This showed how monophenols might influence overall flavour, even at sub-threshold concentrations.

Determination of volatile monophenols in beer using acetylation and headspace solid-phase microextraction in combination with gas chromatography and mass spectrometry.

Monophenols are widely spread compounds contributing to the flavour of many foods and beverages. They are most likely present in beer, but so far, little is known about their influence on beer flavour. To quantify these monophenols in beer, we optimised a headspace solid-phase microextraction method coupled to gas chromatography-mass spectrometry. To improve their isolation from the beer matrix and their chromatographic properties, the monophenols were acetylated using acetic anhydride and KHCO(3) as derivatising agent and base catalyst, respectively. Derivatisation conditions were optimised with attention for the pH of the reaction medium. Additionally, different parameters affecting extraction efficiency were optimised, including fibre coating, extraction time and temperature and salt addition. Afterwards, we calibrated and validated the method successfully and applied it for the analysis of monophenols in beer samples.

Optimisation of specialty malt volatile analysis by headspace solid-phase microextraction in combination with gas chromatography and mass spectrometry.

The objective of this study was to develop a technique for analysing 14 flavour components, relevant for specialty malts. Therefore, a method was developed for the analysis of these components in dry ground malt using headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. A procedure was optimised for the optimal amount of sample, fibre selection, extraction temperature and extraction time. Afterwards, the method was calibrated and validated by the quantification of the specialty malt flavour components in a colour, a caramel and a roasted malt.

Electronic tongue as a screening tool for rapid analysis of beer.

An electronic tongue (ET) comprising 18 potentiometric chemical sensors was applied to the quantitative analysis of beer. Fifty Belgian and Dutch beers of different types were measured using the ET. The same samples were analyzed using conventional analytical techniques with respect to the main physicochemical parameters. Only non-correlated physicochemical parameters were retained for further analysis, which were real extract, real fermentation degree, alcohol content, pH, bitterness, color, polyphenol and CO(2) content. Relationship between the ET and physicochemical datasets was studied using Canonical Correlation Analysis (CCA). Four significant canonical variates were extracted using CCA. Correlation was observed between 6 physicochemical variables (real extract and fermentation degree, bitterness, pH, alcohol and polyphenols' content) and 14 sensors of the ET. The feasibility of the ET for the quantification of bitterness in beer was evaluated in the aqueous solutions of isomerized hop extract and in the set of 11 beers with bitterness varying between 14 and 38 EBU (European Bitterness Units). Sensors displayed good sensitivity to isomerized hop extract and good prediction of the bitterness in beer was obtained. Calibration models with respect to the physicochemical parameters using ET measurements in 50 Belgian and Dutch beer samples were calculated by Partial Least Square regression. The ET was capable of predicting such parameters as real extract, alcohol and polyphenol content and bitterness, the latter with Root Mean Square Error of Prediction (RMSEP) of 2.5.

Decrease of aged beer aroma by the reducing activity of brewing yeast.

The flavor profile of beer is subject to changes during storage. Since, possibly, yeast has an influence on flavor stability, the aim of this study was to examine if there is a direct impact of brewing yeast on aged aroma. This was achieved by refermentation of aged beers. It was shown that several aged aroma notes, such as cardboard, ribes, Maillard and Madeira, were removed almost entirely by brewing yeast, independently of the yeast or the beer type. This was explained by the reduction of aldehydes, mainly (E)-2-nonenal, Strecker aldehydes, 5-hydroxymethylfurfural and diacetyl, to their corresponding alcohols. Furthermore, it became evident that the reducing capacity of brewing yeast is high, but that yeast strain and compound specific residual concentrations remained in the refermented beer independently of the initial concentration. Finally, it appeared that aldehydes were not only reduced but also formed during refermentation.

Instrumental measurement of beer taste attributes using an electronic tongue.

The present study deals with the evaluation of the electronic tongue multisensor system as an analytical tool for the rapid assessment of taste and flavour of beer. Fifty samples of Belgian and Dutch beers of different types (lager beers, ales, wheat beers, etc.), which were characterized with respect to the sensory properties, were measured using the electronic tongue (ET) based on potentiometric chemical sensors developed in Laboratory of Chemical Sensors of St. Petersburg University. The analysis of the sensory data and the calculation of the compromise average scores was made using STATIS. The beer samples were discriminated using both sensory panel and ET data based on PCA, and both data sets were compared using Canonical Correlation Analysis. The ET data were related to the sensory beer attributes using Partial Least Square regression for each attribute separately. Validation was done based on a test set comprising one-third of all samples. The ET was capable of predicting with good precision 20 sensory attributes of beer including such as bitter, sweet, sour, fruity, caramel, artificial, burnt, intensity and body.

Determination of carbonyl compounds in beer by derivatisation and headspace solid-phase microextraction in combination with gas chromatography and mass spectrometry.

Headspace solid-phase microextraction (SPME) followed by gas chromatography and mass spectrometry was applied for quantification of 41 chemically diverse carbonyl compounds in beer. Therefore, in-solution derivatisation with o-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) combined with SPME was optimised for fibre selection, PFBHA concentration, extraction temperature and time and ionic strength. Afterwards, the method was calibrated and validated successfully and extraction efficiency was compared to sampling with on-fibre derivatisation. In-solution derivatisation enabled the detection of several compounds that were poorly extracted with on-fibre derivatisation such as 5-hydroxymethylfurfural, acrolein, hydroxyacetone, acetoin, glyoxal and methylglyoxal. Others, especially (E)-2-nonenal, were extracted better with on-fibre derivatisation.

Decrease of 4-vinylguaiacol during beer aging and formation of apocynol and vanillin in beer.

In this study the decrease of 4-vinylguaiacol (4VG) during beer aging was investigated and the products that arise from it were identified. Two compounds, vanillin and apocynol, were identified in beer model solutions after forced aging and in naturally aged beers by GC-MS and HPLC-ECD analyses. Both account for up to 85% of the decrease of 4VG. Only in the presence of substantial amounts of oxygen in the bottle headspace was vanillin detected. Apocynol [4-(1-hydroxyethyl)-2-methoxyphenol] was found to be the main degradation product, and its formation was shown to be highly dependent on the beer pH. Because both apocynol and vanillin have a clear vanilla-like aroma, the decrease of 4-vinylguaiacol during beer aging might impart a shift from a clove-like aroma in fresh specialty beers (such as wheat beers and other top-fermented blond or dark ales) to a sweeter, more vanilla-like flavor impression of aged specialty beers.

Release and evaporation of volatiles during boiling of unhopped wort.

The release and evaporation of volatile compounds was studied during boiling of wort. The observed parameters were boiling time, boiling intensity, wort pH, and wort density. The effect of every parameter was discussed and approached chemically, with an eye on beer-aging processes. The results indicated that pH highly influenced the release of flavor compounds and that the formation of Strecker aldehydes was linear with boiling time. However, because of evaporation of volatiles, information about the applied thermal load on wort is lost when using a volatile heat load indicator. The thiobarbituric acid (TBA) method, which includes the nonvolatile precursors of volatile aging compounds, proved to be a more reliable method to determine all kinds of heat load on wort. Finally, it was discussed how the obtained insights could help to understand the mechanism of beer aging.

Optimisation of a complete method for the analysis of volatiles involved in the flavour stability of beer by solid-phase microextraction in combination with gas chromatography and mass spectrometry.

Headspace solid-phase microextraction combined with gas chromatography and mass spectrometry was used for the quantification of 32 volatiles which represent the typical chemical reactions that can occur during beer ageing. Detection was accomplished by employing on-fibre derivatisation using o-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) and normal HS-SPME extraction. The procedures were optimised for SPME fibre selection, PFBHA loading temperature and time, extraction temperature and time, and effect of salt addition. Interference of matrix effects was overcome by calibrating according to the standard addition method and by using internal standards. Afterwards, the method was validated successfully and was applied to study the flavour stability of different beer types.

Optimisation of wort volatile analysis by headspace solid-phase microextraction in combination with gas chromatography and mass spectrometry.

The aim of this study was to create a simple, solventless technique without derivatisation in order to analyze a broad range of volatiles in beer wort. A method was developed using headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. The procedure was optimised by selection of the appropriate fibre and optimisation of extraction temperature, extraction time, and salting-out. The detection limits were well below the actual wort concentrations of the selected volatiles, ranging from 12 ng/l for linalool to 0.53 microg/l for furfural. Moreover, the procedure showed a good linearity and was applied to the analysis of wort samples taken from a wort boiling process in an industrial brewery.

Characterization of volatiles in unhopped wort.

The volatile fraction of wort components was studied during boiling. Not less than 118 volatile compounds were identified when unhopped pilsner wort was boiled and samples of wort and condensed vapors were analyzed with headspace SPME-GC/MS, of which 54 were confirmed with reference compounds. The wort samples contained 61 identifiable compounds, while the vapor condensate yielded 108 different compounds. Almost 30 known compounds were found that have not been described before in unhopped pilsner wort. One previously unknown aldol reaction product was tentatively identified as 2-phenyl-2-octenal. The detection of branched 2-alkenals underlines the importance of the aldol condensation in Maillard-type reactions, while the tentative identification of alkyloxazoles and alkylthiazoles could once more accentuate the central role of alpha-dicarbonyl compounds, aldehydes, and amino acids in flavor generation. The condensation of wort vapors joined with the SPME-GC/MS technique has proven to be a useful tool in volatile analysis.