Determination of the Total Soluble Nitrogen Content of Malt and Beer by the Dumas Combustion Method: Collaborative Trial

The determination of the total soluble nitrogen content of malt and beer, by the Dumas procedure, has jointly been collaboratively tested by the Analysis Committee of the Institute of Brewing (IOB) and the European Brewery Convention (EBC). Five samples of beer (range 362 to 1159 mg/l) and five samples of malt (range EBC 0.598 to 0.798 %m/m (dry basis) and IOB 0.534 to 0.706 %m/m (dry basis)) were distributed to eighteen participating laboratories for analysis. Precision values were judged to be independent of the mean soluble nitrogen content for malt by both IOB and EBC methodologies. Values for r₉₅ and R₉₅ were 0.047 and 0.136%m/m for EBC laboratory wort and 0.039 and 0.144 %m/m for IOB laboratory wort respectively. Precision values for beer were judged to be dependent upon the mean nitrogen content (m) in the case of r₉₅ and independent of the mean nitrogen content in the case of R₉₅. Values for r₉₅ and R₉₅ were 0.074m and 120 mg/l respectively.     

ESTIMATION OF ENDO BETA-GLUCANASE ACTIVITY IN MALT USING A VISCOMETRIC METHOD

A viscometric method for estimation of endo beta-glucanase activity in malt has been collaboratively tested by the Institute of Brewing Analysis Committee. Ten participating laboratories analysed five pairs of malt samples in the range 176 to 1238 IRV units. The repeatability (r₉₅) values ranged from 22 to 122 IRV units and the reproducibility (R₉₅) values from 93 to 650 IRV units. The results indicated that R₉₅, but not r₉₅, was dependent on endo beta-glucanase activity. Overall precision values were 76.8 for r₉₅ and 0.506Mean + 4.786 for R₉₅. It was decided that the method would be included in IOB Methods of Analysis but that attention would be drawn to the poor precision values obtained in this and previous collaborative trials.     

DETERMINATION OF THE MOISTURE AND NITROGEN CONTENTS OF BARLEY AND MALT BY NEAR INFRARED SPECTROSCOPY (NIRS)

The determination of the moisture and nitrogen contents of barley and malt by near infrared spectroscopy (NIRS) has been tested by the Analysis Committee of the European Brewery Convention. In the collaborative trial four samples of barley and malt were analysed by 17 laboratories. Repeatability (r₉₅) and reproducibility (R₉₅) values of 0.3 and 1.5% m/m respectively were obtained for barley moisture over the range 12.7 to 15.8% m/m. For malt moisture these values were 0.2 and 1.3% m/m over the range 4.0 to 4.3% m/m, for barley nitrogen 0.1 and 0.3% m/m on dry matter over the range 1.57 to 2.14% m/m, and for malt nitrogen 0.1 and 0.2% m/m on dry matter over the range 1.58 to 1.82% m/m, respectively.     

DETERMINATION OF LOWER BOILING POINT VOLATILE COMPOUNDS IN BEER BY HEADSPACE GAS CHROMATOGRAPHY — COLLABORATIVE TRIAL

The Analysis Committee has collaboratively tested local routine headspace gas chromatographic methods for the determination of the lower boiling point volatile compounds in beer. The repeatability values (r₉₅) were dependent upon mean concentration (m) for acetaldehyde and alcohols but not for esters, whilst reproducibility values (R₉₅) were dependent upon concentration in all cases. The range of values of m and the estimates of r₉₅ and R₉₅ (mg/litre) for each compound were, respectively: acetaldehyde (5–8, 0.21m, 0.60m); propanol (9–23, 0.25m, 2.5+0.57m); isobutanol (5–22, 0.56+0.085m, 1.7+0.15m); methylbutanols (45–105, 0.14m, 0.22m); ethyl acetate (10–54, 3.1, 2.1+0.29m); isoamyl acetate (0.8–4.7, 0.36, 0.20+0.58m); and ethyl hexanoate (0.13–0.36, 0.073, 0.10+0.91m). No advantage was gained by diluting beer samples containing 9% V/V ethanol to 4% ethanol (used for the calibration mixtures) prior to analysis, but use of a standard method of sample preparation decreased most of the R₉₅ values. No recommendation is made in this interim report.     

The Effect of Hydroxycinnamic Acids and Volatile Phenols on Beer Quality

To the consumer, beer should be an agreeable beverage of attractive colour, clarity, pleasing flavour and should carry no toxic substances. One of the major constituents of barley is phenolic acids and their direct effect on the quality of beer is still a mystery. The aim of the study therefore was to investigate the relationship between phenolic acids during mashing at different temperatures and the quality of the beer. The mash regime and liquor to malt ratios were optimised and the carbohydrates obtained were analysed at 3.5 L of water per kg of malt and a total mash time of 105 min. Carbohydrate and phenolic acid analyses were performed by HPLC coupled with a UV‐vis detector. The three different malts used had a phenolic acid content of 33.25 μg/mL, 25.44 μg/mL and 19.98 μg/mL for malt A, B and C, respectively. The plot of 1/T versus Ln(t) gave a negative slope with activation energy (E_a) = 209 KJ/mol, rate constant (k) = 4.6 × 10^?4 mg/L min, which are comparable to similar data reported in the literature. The kinetics studies showed that the optimised mashing temperature of 90°C was adequate to form 4‐vinylguaiacol by thermal decarboxylation from the hydroxycinnamic acids. This study has shown that there is no direct correlation between phenolic acids and oxidative flavour stability of beer while the corresponding volatile phenols may affect beer flavour.     

The influence of unmalted barley on the oxidative stability of wort and beer

The aim of this study was to investigate the influences of unmalted barley on the brewing process and the quality of the resulting beer‐like beverages, with the main focus on the oxidative stability, using traditional beer analyses, GC‐MS for the determination of aging compounds and electron paramagnetic resonance spectroscopy to determine free radical activity. For the investigation, brews with different barley proportions and 75% barley brews with a colour malt addition, to compensate for a lower colour using barley, were produced. In general, it can be said that beers with a proportion of up to 50% barley achieved a comparable or higher extract yield and final attenuation owing to the combined effectiveness of the malt and microbial enzymes. Although all analytical values were within the normal range according to Methodensammlung der Mitteleuropäischen Brautechnischen Analysenkommission (MEBAK), a slight decrease in total polyphenols and free amino nitrogen content was observed. Also in response to higher barley portions, an increase of higher molecular weight proteins and β‐glucan was detected. Barley is not exposed to heat and oxidative stress in the malting plant, which explains the lower values of the thiobarbituric acid index and colour as an indicator of Maillard reaction products in the resulting wort and beer. Additionally, the results demonstrate a slower increase of aging compounds during beer storage with increasing barley proportions. Furthermore, it was observed that higher barley proportions led to a better oxidative stability indicated by a lower radical generation (T₄₅₀‐value) in wort and an increasing beverage antioxidant index/endogenous antioxidative potential (BAX/EAP value) in the final beverage. The case of ‘barley beers’ showed that the positive effect of barley on the oxidative beer stability was greater than the negative effect of the addition of colour malt, to adjust the colour of a 100% malt beer. In sensory comparison with beer produced with 100% malt, the beers brewed with a barley proportion up to 50% showed a slight flavour preference and up to a 75% equivalent evaluation. Copyright © 2012 The Institute of Brewing & Distilling     

COLLABORATIVE DETERMINATION OF SPECIFIC GRAVITY BY DENSITY METER

A method employing a density meter for the determination of specific gravity (SG) has been tested for the determination of the gravity (G) of beer, aqueous sugar solutions and aqueous ethanol solutions by the Analysis Committee of the Institute of Brewing. The term “gravity”, used throughout this report is defined by the equation G = SG × 1000. Repeatability (r₉₅) and reproducibility (R₉₅) values were calculated over the range 994.6 to 1124.2 gravity. For beer, it was judged that precision values were independent of the gravity of the sample. Values for r₉₅ and R₉₅ were 0.1 and 0.9, respectively, for instruments with a 5 figure display and 0.08 and 0.32, respectively, for instruments with a 6 figure display, over the range 999.1 to 1017.0 gravity. For solutions of ethanol in water, precision was also independent of the gravity of the sample. Values for r₉₅ and R₉₅ were 0.1 and 0.3, respectively, for instruments with a 5 figure display and 0.05 and 0.19, respectively, for instruments with a 6 figure display, over the range 994.6 to 999.1 gravity. For aqueous sugar solutions the values of r₉₅ and R₉₅ increased with increasing gravity. At 1049.3 gravity, values for r₉₅ and R₉₅ were 0.1 and 0.8, respectively, for instruments with a 5 figure display and 0.07 and 0.35, respectively, for instruments with a 6 figure display. At 1124.2 gravity values for r₉₅ and R₉₅ were 0.2 and 1.4, respectively, for instruments with a 5 figure display and 0.16 and 1.35, respectively, for instruments with a 6 figure display.     

SPECTROPHOTOMETRIC DETERMINATION OF MALT COLOUR

The International Method for the determination of the colour of beer has been tested by members of the Analysis Committee of the European Brewery Convention on samples of wort produced from a laboratory extract of malt using methods EBC 4.4 and EBC 4.4.5. The method, which relies on the spectrophotometric determination of colour at 430 nm, on clarified worts, is recommended as the designated reference method in place of the current visual method using EBC colour discs. The change will take effect from 1st January 1996. It was judged that precision values were dependent on the intensity of the colour of the sample over the range 3.6 to 25.3 EBC units. Repeatability (r₉₈) and Reproducibility (R₉₈) values of r₉₈ 0.18Mean ? 0.28 and R₉₈ = 0.13Mean + 0.46 were obtained over this range.     

DETERMINATION OF ETHANOL IN BEER BY GAS CHROMATOGRAPHY (DIRECT INJECTION)—COLLABORATIVE TRIAL

A method employing gas chromatography for the determination of ethanol in beer has been collaboratively tested by the Analysis Committee of the Institute of Brewing. It was judged that precision values were independent of concentration over the range 0.93 to 6.05% V/V ethanol. Repeatability (r₉₅) and reproducibility (R₉₅) values of 0.061 and 0.136 respectively, were obtained over this range. At a mean level of 9.17% V/V, the r₉₅ and R₉₅ values were 0.154 and 0.284 respectively. This was probably due to dilution errors as the sample had to be diluted to bring it within the linear range of the method. A comparison of the precision values given by the gas chromatographic method, with those obtained in 1991/1992 by 8 laboratories in a major brewing company using 12 sample pairs, for the IOB Recommended Distillation Method, revealed that there is no significant difference between the precision data for the two methods.     

Determination of the influence of starch sources and mashing procedures on the range of the molecular weight distribution of beer using field‐flow fractionation

To produce experimental beers, different mash mixtures (barley malt, barley malt + 30% pre‐cooked maize, barley malt + 30% nonmalted spelt) and distinct mashing procedures (infusion and decoction) with variations of the rest time and initial temperatures were evaluated. The range of molecular weight distribution (MWD) of the resulting beers was determined using asymmetrical flow field flow fractionation coupled to multiangle laser light scattering and refractive index. There were no differences on the range of MWD among the beers, according to infusion or decoction, using similar raw materials and initial temperatures (45 and 55°C). However the range of MWD was higher (p < 0.005) when using infusion at an initial temperature of 63°C, regardless of the raw material. The use of maize did not alter structural properties of the beer, while mash containing nonmalting spelt caused an elevation on the MWD (p < 0.001) and a lower (p < 0.05) apparent degree of fermentation. Therefore the range of the MWD of the beers was influenced by the quality of the raw material and the initial mashing temperature, whereas apparent degree of fermentation values were affected only by the type of starch source. Thus the determination of the MWD is an important tool for monitoring the production of beer. Copyright © 2013 The Institute of Brewing & Distilling     

DETERMINATION OF REPEATABILITY AND REPRODUCIBILITY OF EBC ACCEPTED METHODS IV—COLOURED MALTS

Methods for the determination of caramel and roasted malt moisture, extract and colour published in Analytica EBC have been collaboratively tested by the European Brewery Convention Analysis Committee, according to the ISO standard 5725. Repeatability (r₉₅) and reproducibility (R₉₅) values are presented.     

The Role of Malt and Hop Polyphenols in Beer Quality,Flavour and Haze Stability

Pilot‐scale brewing trials of a 12°P pale lager beer were conducted to look at the effect of a modified dose of hop and malt polyphenols on haze, flavour quality, and stability. Results confirmed that malt polyphenols, and particularly hop polyphenols, in the course of wort boiling, improved reducing activity values and the carbonyl content in fresh and stored beers. Hop polyphenols significantly increased reducing activity and decreased the formation of carbonyls (TBA value) in fresh and stored beer. Reduced content of malt polyphenols, combined with the use of hop CO₂ extract, caused an increase in the TBA value in beer. PVPP stabilized beers tended to be lower in reducing activity. Both malt and hop polyphenols affected the intensity of “harsh taste” in fresh beers and a significant influence from PVPP stabilization of beer was not observed. The staling degree of forced‐aged beers depended on the polyphenol content in the brewhouse. Both hop and malt polyphenols had a positive impact on flavour stability. PVPP treatment of beer had a positive effect on the flavour stability of heat‐aged beers. Polyphenols, especially hop polyphenols, slowed down flavour deterioration during the nine month storage period, but the primary effect was seen during the first four months of storage. Storage trials did not show any unambiguous effects for PVPP stabilization on beer flavour stability. Results confirmed the negative impact of malt and hop polyphenols on haze stability, and PVPP stabilization minimized differences in shelf life prediction values between beers prepared with the modified dose of polyphenols.     

Evaluation of the fermentative potential of Candida zemplinina yeasts for craft beer fermentation

The fermentative potential of Candida zemplinina Y.01667 and Y.01670 was evaluated to explore the potential use of these yeasts for craft beer fermentations. Fermentation experiments were carried out at different temperatures and soluble solid concentrations, using synthetic media with glucose syrup as a sugar source and with a laboratory malt wort plus different adjuncts. Results showed that both strains fermented well at 14 °C and had improved fermentative activity at 20 °C. The fermentative kinetics of C. zemplinina Y.01667 and Y.01670 were not affected when experiments at higher concentrations of soluble solids were conducted. Furthermore, C. zemplinina strains had better growth, higher viable cells counts, less free amino nitrogen consumption, lower sedimentation rates and slighter changes in pH values, when compared with results of the lager beer yeast Saccharomyces cerevisiae S‐23 in the synthetic medium tested. Fermentations in a malt wort with different adjuncts indicated that C. zemplinina Y.01670 could possibly be used as a yeast in craft beer production. Copyright © 2016 The Institute of Brewing & Distilling     

COLD STERILE FILTRATION: A SMALL SCALE FILTRATION TEST AND INVESTIGATION OF MEMBRANE PLUGGING

Beer brewed from 24 commercially and bag malted samples by a small scale brewing method was assessed by a micro-filtration efficiency (MFE) test designed to emulate the cold-sterile (membrane or micro-) filtration process. The level of malt derived beer components with the potential to reduce MFE, such as β-glucan, arabinoxylan, protein and polyphenol, were consistent over duplicate beer batches suggesting that beer quality was reproducible using the small scale method. The small scale MFE test was able to differentiate (P<0.001) between beer brewed from distinct malt samples in a reproducible fashion, suggesting that the test is effective in assessing beer MFE in the laboratory. Subsequently, the effects of various malt derived beer components on micro-filtration were investigated. MFE (measured as <i>V_max) was negatively correlated with beer arabinoxylan content (r=–0.62, P<0.01), suggesting that the arabinoxylan content of malt, and subsequently beer, may influence MFE. Total beer β-glucan was not significantly related to beer MFE (r=-0.36). However, it was likely that β-glucan molecules of high molecular weight influenced MFE more so than the total β-glucan content. Beer viscosity, which was correlated to both beer β-glucan and arabinoxylan content (r=0.86, P<0.001 and r=0.68, P<0.05, respectively), correlated with V_max (r=-0.81, P<0.001) .     

DETERMINATION OF VICINAL DIKETONES IN BEER BY GAS CHROMATOGRAPHY (HEADSPACE TECHNIQUE)—COLLABORATIVE TRIAL

Two methods for the determination of vicinal diketones in beer have been collaboratively tested by the Analysis Committee of the Institute of Brewing and are recommended for use. Both methods employ gas chromatography and are essentially the same, except that one relies on the use of a packed column and the other on a capillary column. For diacetyl it was judged that repeatability (r₉₅) values were independent of concentration over the range 0.04 to 0.19 mg/litre. Over this range, r₉₅ values for diacetyl of 0.028, 0.020 and 0.026 mg/litre were obtained for capillary, packed columns and combined results respectively. Values for reproducibility (R₉₅) were judged to be dependent on the mean level (m). R₉₅ values were 0.032 + 0.68 m, 0.01 + 0.47 m and 0.005 + 0.67 m were obtained for capillary, packed columns and combined results respectively. For both methods the r₉₅ and R₉₅ values for 2,3-pentanedione were judged to be independent of concentration over the range 0.02 to 0.07 mg/litre. For capillary columns, packed columns and combined results respectively, r₉₅ values were 0.009, 0.009, 0.010 mg/litre and R₉₅ values were 0.037, 0.042 and 0.038 mg/litre.     

ESTIMATION OF ISO-ALPHA-ACIDS IN BEER BY HPLC-COLLABORATIVE TRIAL

A procedure relying on high performance liquid chromatography for the estimation of iso-alpha-acids in beer has been collaboratively tested by the IOB Analysis Committee. In addition the trial samples were analysed by the IOB Recommended Method for the measurement of bitterness (BU). It was judged that the results obtained by the HPLC method were not sufficiently precise to permit adoption as a Recommended Method. However, since the method has the advantage of measuring bitterness in terms of iso-alpha-acids, it is suggested as an alternative to that of the Recommended Method. The iso-alpha-acids are absorbed from beer on to a C₁₈ Bond Elut column and then selectively desorbed prior to isocratic analysis by HPLC using an eluting solvent of methanol/water/phosphoric acid/tetrabutylammonium hydroxide and a C₁₈ radialpak cartridge. For both methods the repeatability values (r₉₅) were not dependent upon mean concentration (m) whereas the reproducibility values (R₉₅) were dependent upon concentration. The values of (r₅₃) and (R₉₅) obtained were 2.11 and (1.38 + 0.134 m) over the concentration range 13.8 to 34.0 mg/litre for the HPLC procedure and 1.20 and (0.76 + 0.122 m) over the concentration range 15.4 to 38.6 BU for the Recommended Method.     

DETERMINATION OF TOTAL NITROGEN IN BARLEY AND MALT BY COMBUSTION METHOD—COLLABORATIVE TRIAL

A combustion method, relying on the Dumas principle, for the determination of total nitrogen in barley and malt, has been collaboratively tested by the Analysis Committee of the European Brewery Convention. Repeatability, r₉₅, and reproducibility, R₉₅, values were 0.063 and 0.116% of dry matter, respectively, for samples with nitrogen contents in the range 1.23 to 1.86% N of dry matter. There was no significant difference between these values for barley and malt. The Analysis Committee approved the adoption of the combustion method for inclusion in Analytica EBC as an alternative method.     

COLLABORATIVE TRIAL ON THE DETERMINATION OF BETA-GLUCANASE IN MALT BY VISCOMETRIC AND DYE-LABELLED METHODS

The results of a collaborative trial to compare a dye-labelled beta-glucan method with that of a viscometric procedure, for estimating the beta-glucanase content of malt, have demonstrated that overall the precisions of both methods are similar. There has been an improvement in the precision of the viscometric method (which is not a Recommended Method) compared with values obtained in previous trials. The Institute of Brewing Analysis Committee judged that the results obtained by the dye-labelled beta-glucan method were not sufficiently precise to permit adoption as a Recommended Method. However, since this procedure has advantages of speed and ease of operation, it is suggested as an alternative method to that relying on viscometry. It is envisaged that a further collaborative trial will be carried out when sufficient experience of the new method has been obtained. The repeatability (r₉₅) and reproducibility (R₉₅) values found were 0.190 m and 0.327 m over the concentration (m,U/kg) range 92–266 U/kg for the dye-labelled method, and 0.173 m and (91.4 + 0.0717 m) over the concentration (m,IRV) range 233–701 IRV units for the viscometric method.     

DETERMINATION OF REPEATABILITY AND REPRODUCIBILITY OF A NEW RAPID ENZYME METHOD FOR THE DETERMINATION OF GLYCOSIDE NITRILE IN MALTED BARLEY

A new rapid method for the determination of malt glycosidic nitrile, using an enzymatic incubation with beta-glucosidase, was tested in an inter-laboratory collaborative trial. Repeatability (r₉₅) and reproducibility (R₉₅) values are reported. A limited comparison is made with the existing determination of malt combined nitrile using methods involving laboratory fermentations.     

The influence of LOX‐less barley malt on the flavour stability of wort and beer

The aim of this study was to investigate the influence of lipoxygenase‐less (LOX‐less) barley malt on the quality of wort and beer, with the main focus on beer flavour stability. In the current study, pilot‐scale (1000 L) brewing trials were conducted with a control barley malt AC Metcalfe and a LOX‐less barley malt, PolarStar. The results clearly indicated that the LOX‐less barley malt showed less nonenal potential than the control, although LOX activities in both barley malts were relatively low. The beer brewed from the LOX‐less barley malt contained much lower concentrations of trans‐2‐nonenal (T2N) and gamma‐nonalactone, especially after the (forced or natural) aging of the beer, compared with the beer brewed under the same conditions using the control malt. The sensory panel evaluation indicated similar results in the general flavour profile. The freshness scores of beer brewed from the LOX‐less malt were higher than those from the control malt, and this was more pronounced after forced aging. In addition, the beer brewed from LOX‐less malt had a much better foam stability, almost 30 s (NIBEM test). These results confirm that the use of the LOX‐less barley malt was beneficial to beer flavour stability and foam stability. Copyright © 2014 The Institute of Brewing & Distilling