Analysis of caramels by capillary electrophoresis and ultrafiltration

Class I, III and IV caramels are distinguished from each other by capillary electrophoresis at pH 2.5 and 9.5. The majority of the colour is shown to be in the high molecular weight (MW) fraction of all the caramels. The high MW Class I and Class IV caramel peaks migrate with a negative charge at both pHs but the Class IV caramel also has several sharp peaks from low MW components compared to only a small neutral peak from the Class I caramels. The Class III caramels have a high MW peak which is positively charged at pH 2.5. The migration time, and hence charge, of the high MW peak of Class III caramels is shown to be related to the caramel's nitrogen content. © Crown copyright 1999     

Barley and Malt Proteins and Proteinases: I. Highly Degradable Barley Protein Fraction (HDBPF), a Suitable Substrate for Malt Endoprotease Assay

Barley grain proteins were extracted and fractionated, based on their solubility, to investigate their proteolytic digestibility and suitability to be used as a substrate for the assay of malt proteinases. The fraction extracted with alkaline buffer (Tris‐HCl or Tris‐glycine pH 8.8–9.5), at the end of the sequence, exhibited remarkably high degradability by malt proteases compared to other fractions or any known protein substrate. Gel filtration chromatographic analysis of this fraction revealed that it is composed of three different molecular weight components. Further investigation, after proteolytic treatment, demonstrated that the third and the low molecular weight component is the highly degradable protein(s) (HDP). We designated the whole fraction, the mixture of the three components, as the highly degradable barley protein fraction (HDBPF) and used it (and recommend it) as the substrate for the assay of malt endoproteases activity.     

Effect of High Temperature‐High Humidity Treatment of Germinated Unkilned Barley on Malt Quality and Extract Characteristics

The effect of a high temperature‐high humidity treatment (HT‐HHT) of germinated unkilned barley on malt quality and extract characteristics was studied. Two samples of six‐row barley were steeped to 42% moisture and germinated, with and without gibberellic acid, at 15°C for 5 days. The germinated barley was placed in a high humidity (75–80%) atmosphere maintained at 45, 55, and 65°C, respectively. For each temperature, treatments were carried out for 30, 60 and 90 min, respectively. At 45°C for 30–60 min, the malts developed high diastatic power and proteolytic activity. The high values for cold water extract and reducing sugars in the extracts indicated extensive amylolysis of starch granules during HT‐HHT of the germinated barley at 55–65°C. The worts were light in colour, with a pH of 5.3–5.8 and titratable acidity was in the range of 0.09‐0.23%. A consistent increase in soluble nitrogen and Kolbach index was observed in the malts treated at 45–55°C for 30–90 min. Free α‐amino nitrogen of the malts was in the desirable range of 120–150 mg L^?1. Therefore, HT‐HHT can be useful for improving malt modification and wort characteristics and to shorten the germination time for malts from poor quality barley.     

EXAMINATION OF SOME BREWING MATERIALS AND CARAMELS FOR THE POSSIBLE OCCURRENCE OF 4-METHYLIMIDAZOLE

An investigation was carried out to determine how much, if any, 4-methylimidazole is present in caramels and some other materials used in brewing. Existing analytical methods were modified to improve sensitivity. 4-Methylimidazole was not detectable in roasted barley, chocolate malt, crystal malt, Farbebier, wort and beer, when the limit of detection was 1 mg/kg. However, caramels manufactured by different processes contained amounts ranging from undetectable to 280 mg/kg.     

Utilization of the Linear Mode of MALDI‐TOF Mass Spectrometry in the Study of Glycation During the Malting Process

The process of glycation during the malting process was monitored by the linear mode of matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐TOF MS). Water‐soluble proteins were investigated and two hulled barley varieties, Jersey and Tolar, were compared to the hulless line KM 1910. The crude extracts of the proteins obtained from the grain, the malt, and aliquots collected every 24 h during the malting process, were mixed with the matrix (2,6‐dihydroxyacetophenone) and analyzed by mass spectrometry. The protein composition of the barley changed during the malting process. The protein patterns did not differ significantly between the three varieties of the barley grains. However, significant differences between the malts were evident. Results showed the influence of the malting process on the glycation of certain water‐soluble barley proteins, nonspecific lipid transfer protein 1 (LTP1) and protein Z, of which the glycated forms survived the brewing process. These major barley proteins are very important for the formation and stability of beer foam and glycation may prevent their precipitation. Analysis results indicated that slight glycation of the proteins had occurred on the second day of malting. The linear mode of MALDI‐TOF mass spectrometry was used as a fast and simple method for monitoring the patterns of low‐molecular weight barley proteins with regard to barley variety discrimination. This procedure also enables the selection of barley varieties suitable for the malting industry.     

THE INSTITUTE OF BREWING ANALYSIS COMMITTEE REVISED METHODS OF EXTRACT DETERMINATION

Following a Brewer's Society request to consider a change from a standard Miag Disc Mill setting of 0·5 mm to one of 0·7 mm the Analysis Committee has re-evaluated the determination of hot water extract. It is recommended that from 1st December 1979–1. A Miag Disc Mill setting of 0·7 mm be adopted for grinding malt for the determination of hot water extract. 2. Where an estimate of the fine/coarse grind hot water extract difference is required, mill settings of 0·2 mm and 0·7 mm respectively should be used. The reproducibility of this procedure is adequate to separate malts into classes with high and low values but analytical tolerances cannot be recommended for commercial transactions. Revised methods for setting the Miag Disc Mill and for determining the hot water extract of fine and coarse ground malts are given in Appendix I. It is proposed that all mashes should be made in stainless steel beakers, the use of the standard brewers' mash flask being limited to making the mash to the correct volume at the end of the determination. It is recommended that, from 1st December 1979, the 0·7 mm Miag Disc Mill setting should replace also the 0·5 mm setting in all Recommended Methods for coloured malts, dark malts, roasted barley and unmalted grain. It is further recommended that from 1st December 1979 all specific gravity determinations for laboratory extracts of malt, coloured malt, dark malt, roasted barley, unmalted grain, sugars, syrup, caramel and spent grains should be made at 20°C.     

The role of ferulic acid and arabinoxylan in inducing premature yeast flocculation

Premature yeast flocculation (PYF) is a universal phenomenon and has caused serious problems in the brewing industry. For brewing quality control, it is of great value to investigate the PYF factors that induce this destructive phenomenon. In the present study, two barley malts (PYF+ and PYF?), made from the same barley cultivar by one maltster in China, were selected for PYF factor investigations. The results showed that considerably higher amounts of the bound ferulic acid (FA), rather than arabinoxylan (AX), existed in the PYF+ wort compared with the PYF? wort. To better understand the role of these polysaccharides in PYF+ and PYF? wort, they were fractional precipitated with graded ethanol concentrations. The AX and FA contents in each fraction, as well as the molecular weight profiles and monosaccharide composition, were determined. Furthermore, the PYF?inducing activities of each fraction were measured using a small‐scale fermentation. It was found that the 40% ethanol‐precipitated fraction of both the PYF+ and PYF? wort, and the 60% fraction from the PYF+ wort, could induce the severe PYF phenomenon when added to the PYF? wort. These fractions were proposed to contain PYF factors of suitable molecular weight and a sufficient amount of the polysaccharides. In addition, it was found that the bound FA in arabinoxylan behaved as an important PYF factor in barley malt. The results of this work may contribute to the further study of the PYF mechanism. Copyright © 2015 The Institute of Brewing & Distilling     

THE USE OF HORDEIN FRACTIONS TO ESTIMATE PROTEOLYTIC ACTIVITY IN BARLEY AND MALT

When the natural barley protein, hordein, is used as a substrate for the assay of exopeptidase and endopeptidase activity in green malts, the enzymic activities found differ significantly from those obtained using non-barley substrates such as dipeptides, gelatin or haemoglobin. Malts having similar total levels of carboxypeptidases as measured using non-barley substrates showed considerable variation in the extent to which these enzymes break down barley hordein. Malts also varied more in their endopeptidase activity with hordein than in their ability to digest gelatin or haemoglobin. Analyses of hordein by gel chromatography and gel electrophoresis indicate that it consists of eight main components, which can be divided into three groups of proteins, with molecular weights in the regions of 15,000, 65,000 and ≥ 100,000. Barley samples differed in their relative contents of these proteins and this variation could be rapidly assessed using gel electrophoresis.     

α-GLUCOSIDASE ACTIVITY OF SORGHUM AND BARLEY MALTS

Sorghum malt α-glucosidase activity was highest at pH 3.75 while that of barley malt was highest at pH 4.6. At pH 5.4 employed in mashing sorghum malt α-glucosidase was more active than the corresponding enzyme of barley malt. α-Glucosidase was partly extracted in water but was readily extracted when L-cysteine was included in the extraction buffer, pH 8. Sorghum malt made at 30°C had higher α-glucosidase activities than the corresponding malts made at 20°C and 25°C. Nevertheless, the sorghum malts made at 20°C and 25°C produced worts which contained more glucose than worts of malt made at 30°C. Although barley malts contained more α-glucosidase activity than sorghum malts, the worts of barley had the lowest levels of glucose. The limitation to maltose production in sorghum worts, produced at 65°C, is due to inadequate gelatinization of starch and not to limitation to β-amylase and α-amylase activities. Gelatinization of the starch granules of sorghum malt in the decantation mashing procedure resulted in the production of sorghum worts which contained high levels of maltose, especially when sorghum malt was produced at 30°C. Although the β-amylase and α-amylase levels of barley malt was significantly higher than those of sorghum malted optimally at 30°C, sorghum worts contained higher levels of glucose and equivalent levels of maltose to those of barley malt. It would appear that the individual activities of α-glucosidase, α-amylase and β-amylase of sorghum malts or barley malts do not correlate with the sugar profile of the corresponding worts. In consequence, specifications for enzymes such as α-amylase and β-amylase in malt is best set at a range of values rather than as single values.     

国产大麦蛋白质含量及酶活力变化对麦芽品质的影响

选用蛋白质含量不同的6种国产大麦,五种蛋白组分含量,蛋白酶活力变化及对其制成麦芽理化指标进行了研究。国产大麦蛋白含量较高,各蛋白组分因品种不同差异较大,参与大麦代谢活动的组分波动较大,贮存蛋白变化相对平稳,研究表明大麦蛋白酶活力与五种蛋白组分含量不具有相关性,而制成麦芽后,麦芽蛋白酶活力与麦芽泡沫蛋白成负相关r=-0.733(p<0.05),与麦芽盐溶蛋白成负相关r=-0.789(p<0.05),与麦芽库值成正相关r=0.694(p<0.05)。     

Quality of pilsner malt and roasted malt during storage

Malt is usually expected to be stable during 12 months of storage. However, in practice many brewers notice changes in malt aroma during storage. The oxidative stabilities of pilsner malt and roasted malt were evaluated during a 12 month storage at different temperatures (10 and 20 °C) and water activities (0.231 and 0.432). The radical content in malt kernels was measured by electron spin resonance spectroscopy and the volatile profile of the resulting sweet worts was measured by head‐space analysis followed by GC‐MS analysis. The storage of malt resulted in oxidative reactions and a large change of the volatile profile of the resulting worts. Roasted malt was much more unstable than pilsner malt, as illustrated by a higher initial radical intensity, larger radical decay during storage and a larger change in the volatile profile of the wort with increased amounts of lipid oxidation products. For both roasted malt and pilsner malt, good correlations were found between radical decay and changes in the volatile profile of the wort, where high temperature and high water activity resulted in the largest changes. During the 12 months of storage, the sugar extract of the wort made from the malts remained constant and was not affected by the chemical changes. This study suggests that chemical changes occurring in malts during less than 12 months of storage may potentially affect the aroma of beer, and that water activity and storage temperature should both be kept low in order to maintain a high malt quality. Copyright © 2014 The Institute of Brewing & Distilling     

Effects of nitrogen application on malt modification and dimethyl sulfide precursor production in two Japanese barley cultivars

BACKGROUND: Nitrogenous components have a great influence on both malt and beer qualities. Barley storage proteins are degraded during the germination process, in which amino acids and small peptides are released. Some of these compounds relate to dimethyl sulfide precursor production in the malting process. In this study, barley and malt qualities were investigated using two Japanese barley cultivars, Sukai Golden and Mikamo Golden, with several different nitrogen (N) treatments. RESULTS: Nitrogen top‐dressing treatments efficiently increased N and sulfur (S) concentrations in grains. A difference in malt modification was induced by these treatments without any change in protease activity in malts. S‐Methyl methionine (SMM) concentration in malt of Sukai Golden with low‐N treatment was 1.8–2.1 times higher than that with higher‐N treatments. Methionine concentration in malts was not significantly affected by N treatments of both cultivars, while grain S level was not consistent under any treatments. CONCLUSION: Results show that low‐N treatment increases SMM concentration in malts despite major S‐containing amino acids of malts being not highly affected by the difference in nutrient status of grains. Further investigations are necessary into aspects of both metabolic profiles in barley germination and SMM degradation in the kilning process. Copyright © 2008 Society of Chemical Industry     

Quality characteristics of triticale malts and worts

Worts from triticale malts, in particular well modified malts, separated poorly from mashes. Worts prepared at 70° C had high viscosities (10–27 cSt) indicating that problems would occur during filtration in brewing. The viscosities of triticale worts were higher than those of worts from barley malts. In addition, worts from well modified malts were generally turbid. Proteinaceous material (partly degraded prolamins) was the primary cause of this turbidity. Although the degree of malt modification did influence the rate of wort separation, it had little effect on wort viscosity. High viscosity was caused by pentosans dissolved from the triticale malt during mashing. Oxidative gelation was not observed with these pentosans. Grains and malts were fractionated, and the high molecular weight fractions were analysed for their sugar and acyl components. All were rich in arabinose and xylose. There was a rough inverse correlation between the solubility of the poly saccharide fractions and the levels of substitution with acetyl and feruloyl residues. The poor wort separation from triticale malt grists appeared to be related to the particle size distributions, which were narrow. The sedimentation values of the grist ‘fines’ were high.     

Optimised quantification of the antiyeast activity of different barley malts towards a lager brewing yeast strain

The brewing of beer involves two major biological systems, namely malted barley (malt) and yeast. Both malt and yeast show natural variation and assessing the impact of differing malts on yeast performance is important in the optimisation of the brewing process. Currently, the brewing industry uses well-established tests to assess malt quality, but these frequently fail to predict malt-associated problem fermentations, such as incomplete fermentations, premature yeast flocculation (PYF) and gushing of the final beer product. Antimicrobial compounds, and in particular antiyeast compounds in malt, may be one of the unknown and unmeasured malt factors leading to problem fermentations. In this study, the adaptation of antimicrobial assays for the determination of antiyeast activity in malt is described. Our adapted assay was able to detect differing antiyeast activities in nine malt samples. For this sample set, malts associated with PYF during fermentation and gushing activity in beer showed high antiyeast activity. Both PYF and gushing are malt quality issues associated with fungal infection of barley in the field which may result in elevated antimicrobial activity in the barley grain. Also, two more malts that passed the normal quality control tests were also observed to have high antiyeast activity and such malts must be considered as suspect. Based on our results, this assay is a useful measure of malt quality as it quantifies the antiyeast activity in malt which may adversely impact on brewery fermentation.     

Antioxidant Properties of Free,Soluble Ester and Insoluble‐Bound Phenolic Compounds in Different Barley Varieties and Corresponding Malts

Ten different barley cultivars and their corresponding malts were used to obtain different fractions. Phenolics extracted belonged to free, soluble esters and insoluble‐bound fractions. Total phenolic content (TPC) of the free fraction, as measured according to the Folin‐Ciocalteu method, ranged from 37.7 to 167.2 mg gallic acid equiv/kg of dried material (GAE/kg_dw) for barley and between 34.1 and 72.3 mg GAE/kg_dw for malt. The bound phenolic content ranged from 210.3 to 320.5 and between 81.1 and 234.9 mg GAE/kg_dw for barley and malt, respectively. The contribution of bound phenolics to the TPC was significantly higher than that of free and esterified fractions. Catechin and ferulic acid, quantified by high performance liquid chromatography with diode array detector (HPLC‐DAD), were the most abundant phenolics in the free and bound fractions, respectively. The p‐coumaric acid content was lower in hulless genotypes, as compared to hulled genotypes, showing that it is mainly concentrated in the hull. The antioxidant activities of the phenolic fractions were investigated using the radical scavenging assay (DPPH) and ferricyanide reducing power. The bound phenolics demonstrated a significantly higher antioxidant capacity compared to the free and esterified phenolics. During the malting process, a significant decrease of the bound phenolics was observed with a corresponding increase of the esterified fraction.     

125th Anniversary Review: The science of the tropical cereals sorghum,maize and rice in relation to lager beer brewing

Mainstream lager beer brewing using the tropical cereals sorghum, maize and rice, either as malt or as raw grain plus commercial enzymes, is becoming widespread. This review examines the differences in composition between these tropical cereals and barley and their impact on brewing processes and beer quality. All of these cereals have a starch gelatinization temperature some 10 °C higher than barley. The sorghum prolamin proteins are particularly resistant to proteolysis owing to disulphide cross‐linking involving γ‐kafirin. Unlike barley, the major endosperm cell wall components in sorghum and maize are arabinoxylans, which persist during malting. The rice cell walls also seem to contain pectic substances. Notably, certain sorghum varieties, the tannin‐type sorghums, contain considerable levels of condensed tannins (proanthocyanidins), which can substantially inhibit amylases, and probably also other brewing enzymes. Tropical cereal malts exhibit a similar complement of enzymic activities to barley malt, with the notable exception of β‐amylase, which is much lower and essentially is absent in their raw grain. Concerning beer flavour, it is probable that condensed tannins, where present in sorghum, could contribute to bitterness and astringency. The compound 2‐acetyl‐1‐pyrroline, responsible for the popcorn aroma of maize and also the major aroma compound in rice, presumably affects beer flavour. However, much more research is needed into tropical cereals and beer flavour. Other future directions should include improving hydrolysis of prolamins into free amino nitrogen, possibly using prolyl carboxypeptidases and investigating tropical cereal lines with useful novel traits such as high amylopectin, high protein digestibility and low phytate. Copyright © 2013 The Institute of Brewing & Distilling     

Identification and quantification of Class IV caramels using capillary electrophoresis and its application to soft drinks

A capillary electrophoresis method was developed for the analysis of Class IV caramels using sodium carbonate buffer at pH 9·5. A relationship was established between the migration time of the coloured ‘caramel’ peak and the caramel's sulphur content. There was also a linear relationship between the concentration of caramel solids in water and the peak area of the ‘caramel’ peak in the range 0·1–10 g litre⁻¹. A range of soft drinks was analysed for caramel. Seven non-diet and 12 diet cola products had Class IV caramel of high-sulphur, high-nitrogen content at concentrations estimated to be between 0·57 and 0·82 g solids litre⁻¹. © Crown Copyright 1998.     

MALTING AND BREWING WITH BARLEYS HAVING BLUE ALEURONES

Blue aleurones in barley are associated with elevated levels of polyphenolic materials such as anthocyanins and anthocyanidins. A rapid method has been developed for assessing the anthocyanin content of barleys, malts and worts. Malts were prepared from a range of barleys, some of which had intensely blue aleurones, while others were only slightly blue or showed no visible pigmentation. The malting quality of barley was not affected by aleurone colour and ales and lagers of sound flavour as well as acceptable analytical parameters were brewed from malts having pronounced blue aleurones. In some cases sweet worts prepared from blue aleurone malts had a slight pinkish tinge, but this disappeared during wort boiling, and beer colours were normal. Levels of anthocyanins in barley correlated with those in malt and in wort. However, the concentration of anthocyanins was unrelated to the amount of anthocyanogens or total polyphenols. High anthocyanin levels in either barley or beer had no deleterious effects on beer flavour or the rate of haze development.     

Determination of antioxidants in edible grain derivatives from the Canary Islands by capillary electrophoresis

A preliminary study of the antioxidant content of hand-produced grain derivatives from the Canary Islands, named gofio, is developed by means of a capillary electrophoresis (CE) method. Protocatechuic acid, salicylic acid, p-hydroxybenzoic acid, vanillic acid, syringic acid, p-coumaric acid, ferulic acid and sinapic acid were extracted from corn, wheat, barley and pea gofio samples by sonication with methanol and were successfully separated by CE using a polycation, hexadimetrine bromide (HDB), as electrosmotic flow modifier. The separation was achieved using a running buffer consisting of 125 mM boric acid, 49 mM disodium hydrogen phosphate, 0.002% (w/v) HDB and 2.5 mM α-cyclodextrin at pH 7.5. The developed procedure allowed the simultaneous determination of the selected antioxidants in less than 3.5 min. The electrophoretic profile obtained for each kind of flour (corn, wheat, barley and pea) allowed differentiation of the type of gofio. Among the analyzed samples, corn gofio showed the highest antioxidant content. Furthermore, highly roasted corn or wheat gofio samples were also analyzed and their antioxidant contents were lower than those of less roasted samples.     

PURIFICATION OF BARLET MALT α-AMYLASE BY IMMUNOAFFINITY CHROMATOGRAPHY

Immunoaffinity chromatography was used to purify the high pl α-amylase (α-amylase II) in a one step procedure after fractionation of the whole barley malt extract on Sephadex G25. The immunoglobulin G (IgG) fraction of an immune serum specific for the malt α-amylase II was immobilized on Ultrogel. A mild desorption procedure was used, combining distilled water elution with an interrupted elution. The purification was achieved within half a day including kernel extraction. The quality of the purification was assayed by SDS polyacrylamide gel electrophoresis, crossed immunoelectrophoresis and isoelectric focusing. For the second technique, an immune serum was used which was polyspecific for malt proteins including the high pl α-amylase (α-amylase II). The effect of this procedure on the specific activity of the enzyme and on its antigenicity was evaluated. The results underline the efficiency of the purification procedure and indicate that α-amylase II accounts for a few percent of the total soluble protein in malts. However, the α-amylase II fraction was not completely free from α-amylase I. The procedure resulted in a partial loss of the enzymatic activity but not of the antigenicity.