Multivariate analysis of NMR and FTIR data as a potential tool for the quality control of beer

In this work, principal component analysis (PCA) is applied to the FTIR-ATR and the (1)H NMR spectra of 50 beers differing in label and type (ale, lager, alcohol-free), to identify the spectral parameters that may provide rapid information about factors affecting beer production. PCA of FTIR data resulted in the separation of beers mainly according to their alcoholic content, providing little information on components other than ethanol contributing to the variability within the samples. PCA of (1)H NMR spectra, performed on the region where major beer components resonate (3.0-6.0 ppm), resulted in the separation of samples into four groups: two groups characterized by the predominance of dextrins, one group of alcohol-free beers characterized by the predominance of maltose, and one group where glucose was found to predominate. By performing PCA on aliphatic and aromatic regions, the contribution of minor components was highlighted. In particular, most ales, lagers, and alcohol-free samples could be distinguished based on their aromatic composition, thus reflecting the high sensitivity of the low-field NMR region toward different types of beer fermentation.     

Phenol antioxidant quantity and quality in foods: fruits.

The free and bound phenols have been measured in 20 fruits commonly consumed in the American diet. Phenols were measured colorimetrically using the Folin-Ciocalteu reagent with catechin as the standard after correction for ascorbic acid contribution. On a fresh weight basis, cranberry had the highest total phenols, and was distantly followed by red grape. Free and total phenol quality in the fruits was analyzed by using the inhibition of lower density lipoprotein oxidation promoted by cupric ion. Ascorbate had only a minor contribution to the antioxidants in fruits with the exception of melon, nectarine, orange, white grape, and strawberry. The fruit extracts' antioxidant quality was better than the vitamin antioxidants and most pure phenols, suggesting synergism among the antioxidants in the mixture. Using our assay, fruits had significantly better quantity and quality of phenol antioxidants than vegetables. Fruits, specifically apples and cranberries, have phenol antioxidants that can enrich lower density lipoproteins and protect them from oxidation. The average per capita consumption of fruit phenols in the U.S. is estimated to be 255 mg/day of catechin equivalents.     

Identification of a stale-beer-like odorant in extracts of naturally aged beer

For a long time, beer staling has been a prime concern in brewery research. Yet, to improve flavor stability, better knowledge of all chemicals involved is still needed. From our aroma extract dilu-tion analyses (AEDA) applied to naturally aged lager beers emerged an old-beer-like odorant at RICP-SIL 5 CB = 1532 and RIFFAP = 2809, with a FD value close to that of trans-2-nonenal (the well-known cardboard off-flavor found in aged beers). Specific phenol extraction, GC cold trapping, and mass spectrometry (electron impact and chemical ionization) enabled us to identify it as 4-vinylsyringol. Although already mentioned in some fresh beers, this compound had never been highlighted as involved in the aging process of lager beers.     

Heavy and light beer: a carbon isotope approach to detect C(4) carbon in beers of different origins,styles, and prices

The carbon isotope ratios (delta(13)C) of 160 beers from around the world ranged from -27.3 to -14.9 per thousand, primarily due to variation in the percentage of C(3) or C(4) plant carbon in the final product. Thirty-one percent of beers had a carbon signature of C(3) plants (barley, rice, etc.), whereas the remaining 69% contained some C(3)-C(4) mixture (mean of mixtures, 39 +/- 11% C(4) carbon). Use of C(4) carbon (corn, cane sugar, etc.) was not confined to beers from any particular region (Pacific Rim, Mexico, Brazil, Europe, Canada, and the United States). However, the delta(13)C of European beers indicated mostly C(3) plant carbon. In contrast, U.S. and Canadian beers contained either only C(3) or C(3)-C(4) mixtures; Brazilian, Mexican, and Pacific Rim beers were mostly C(3)-C(4) mixtures. Among different lagers, U.S.-style lagers generally contained more C(4) carbon than did imported pilsners. Among different ales, those brewed by large high-production breweries contained significant proportions of C(4) carbon, while C(4) carbon was not detected in microbrewery or home-brew ales. Furthermore, inexpensive beers generally contained more C(4) carbon than expensive beers.     

High-resolution nuclear magnetic resonance spectroscopy and multivariate analysis for the characterization of beer

Beer contains a very complex mixture of nutrients, which in this work are identified to some extent by high-field high-resolution nuclear magnetic resonance (NMR) one- and two-dimensional methods. The (1)H NMR spectrum of beer shows a predominance of strongly overlapped peaks arising from several carbohydrates. Minor components are clearly observed both in the aliphatic and in the aromatic regions of the spectrum. With the aid of two-dimensional methods, spectral assignment was carried out, enabling the identification of approximately 30 compounds and identifying about the same number of spin systems for further assignment. The variability of the spectral profile of beers differing in type and label was studied by principal component analysis (PCA), and it was found that, although some distinction is achieved on the basis of the aliphatic and sugar compositions, clearer separation between ales and lagers is obtained by PCA of the aromatic profiles alone. The potential of this technique as a rapid and informative quality control tool is discussed.     

Spectrophotometric method for exploring 3-methyl-2-butene-1-thiol (MBT) formation in lager

The disappearance of riboflavin absorbance at 445 nm from beers or model beers on light exposure is directly linked to light-struck character formation. The addition of (+)-catechin, (-)-epicatechin, tryptophol, or ascorbic acid was able to reduce, but not stop, absorbance loss or light-struck character formation in either model beer or mainstream lager that was exposed to light. When isohumulone was present in model beer, the inhibitory effect of (+)-catechin, (-)-epicatechin, or tryptophol decreased with increasing isohumulone. The spectrophotometric method used in this study is a simple and effective method for determining light-struck susceptibility.     

Determination of estrogenic activity in beer by biological and chemical means

It has been suspected that beer drinking may change the hormonal status of men caused by phytoestrogens. Five different Austrian lager beers have been investigated for estrogenic activity by a yeast two-plasmid system harboring the human estrogen receptor alpha, after concentration by solid phase extraction. The beer concentrate was further fractionated by reversed phase HPLC, and then the fractions were characterized by the biological assay and GC-MS. The most potent fraction did not contain a known phytoestrogen. The total activity corresponded to an average of 43 ng of 17beta-estradiol/L of beer. It was concluded that the human health hazard of beer drinking originating from compounds activity on the estrogen receptor alpha is negligible.     

Exploration of beer proteome using OFFGEL prefractionation in combination with two-dimensional gel electrophoresis with narrow pH range gradients

Two-dimensional gel electrophoresis in combination with mass spectrometry has already been applied successfully to study beer proteome. Due to the abundance of protein Z in beer samples, prefractionation techniques might help to improve beer proteome coverage. Proteins from four lager beers of different origins were separated by two-dimensional electrophoresis (2-DE) followed by tandem mass spectrometric analysis. Initially 52 proteins mostly from Hordeum vulgare (22 proteins) and Saccharomyces species (25 proteins) were identified. Preparative isoelectric focusing by OFFGEL Fractionator was applied prior to 2-DE to improve its resolution power. As a result of this combined approach, a total of 70 beer proteins from Hordeum vulgare (30 proteins), from Saccharomyces species (31 proteins), and from other sources (9 proteins) were identified. Of these, 37 proteins have not been previously reported in beer samples.     

The quantitative analysis of thiamin and riboflavin and their respective vitamers in fermented alcoholic beverages

This research aimed to develop a simple and effective method for analyzing thiamin (B(1)), riboflavin (B(2)) and their respective vitamers by high performance liquid chromatography (HPLC) in fermented alcoholic beverages. The method developed here employs a phosphate buffer/methanol gradient elution on a single reverse phase column, coupled with independent fluorescent detection regimes. It also employs a precolumn derivatization to convert thiamin to thiochrome via an alkaline potassium ferricyanide solution. The method described here allowed a spike recovery of better than 97%, with a typical linear detection range (R(2) ≥ 0.9997) between ≤ 5 and ≥ 500 μg/L for all vitamers studied. Lager style beers were found to contain significantly (p < 0.001) less thiamin than other tested styles of beers (lager, 35.7 μg/L; ale, 88.3 μg/L; stout/porters, 104.4 μg/L; wheat beers, 130.7 μg/L), which may be due to the raw material and extensive processing that occurs for this style. There was no statistical difference (p = 0.608) between the riboflavin content of each beer style. Furthermore, wines and ciders contain less thiamin and riboflavin than beer, which is also likely to be due to the base materials used and the differences in processing steps to produce these beverages.     

Modification of the levels of polyphenols in wort and beer by addition of hexamethylenetetramine or sulfite during mashing.

The effects of addition of hexamethylenetetramine (HMT) or sulfite during mashing on the polyphenol content and oxidative stability of wort and beer have been evaluated in a series of laboratory mashings and pilot brews. HMT reduced the concentration of catechin, prodelphinidin B-3, and procyanidin B-3 in wort and beer, whereas the concentration of ferulic acid was unaffected. Sulfite had only a minor effect on the concentration of phenolics in wort and beer. Addition of HMT or sulfite during mashing increased the oxidative stability of the beer slightly as judged by the tendency of formation of radicals (ESR spin trapping technique), although sensory analysis gave identical flavor acceptance scores to beers produced from untreated and HMT-treated wort and lower scores to beer from sulfite-treated wort. No difference in the oxidative stability of the differently treated sweet worts could be detected as judged by the rate of formation of radicals. HMT addition during mashing has thus been demonstrated to be a valuable experimental tool to control the level of polyphenols in wort and for producing brews with various levels of polyphenols from a single malt.     

Chocolate is a powerful ex vivo and in vivo antioxidant, an antiatherosclerotic agent in an animal model, and a significant contributor to antioxidants in the European and American Diets

Chocolate today is often viewed as a food or snack with little nutritional value. The high saturated fat content of chocolate has also contributed to the belief that its consumption increases the risk of heart disease. However, recent human studies have proven that chocolate has beneficial effects on some pathogenic mechanisms of heart disease such as endothelial function and blood pressure. Although the antioxidant properties of chocolate have been known for some time, there has been no examination of its place in the U.S. diet as a source of antioxidants. This paper demonstrates that chocolate makes a significant contribution to U.S. per capita dietary antioxidants and by inference the European Community's. In the U.S. diet chocolate is the third highest daily per capita antioxidant source. An ex vivo study shows that epicatechin, a major polyphenol in chocolate and chocolate extracts, is a powerful inhibitor of plasma lipid oxidation due to polyphenols' ability to bind to lower density lipoproteins. Conversely, the fat from chocolate alone is a pro-oxidant in this model. This is also demonstrated in an in vivo human study. After consumption of dark chocolate and cocoa powder, the lower density lipoproteins isolated from plasma were protected from oxidation compared to the lipoproteins isolated after cocoa butter consumption, which were put under oxidative stress. In an animal model of atherosclerosis, cocoa powder at a human dose equivalent of two dark chocolate bars per day significantly inhibited atherosclerosis, lowered cholesterol, low-density lipoprotein, and triglycerides, raised high-density lipoprotein, and protected the lower density lipoproteins from oxidation. Chocolate has thus been shown to have potential beneficial effects with respect to heart disease.     

Involvement of flavanoids in beer color instability during storage

Besides Maillard reactions, structural rearrangements of flavan-3-ol monomers cause color changes in beer during storage. Acetone/water-soluble fractions (70/30, v/v) of three lager beers of the same batch, differently stabilized before bottling in glass or poly(ethylene terephthalate) (PET) bottles, were monitored by normal-phase HPLC-ESI(-)-MS/MS over a 1-year period of storage at 20 degrees C. In parallel, beer color was monitored by the European Brewery Convention assay. The evolution of color was similar in the silica gel-filtered beer to that in identically bottled and stored poly(vinylpolypyrrolidone)-treated samples, despite the high flavanoid dimers content of the former. On the other hand, color evolved more rapidly in the PET bottle, suggesting a key role of oxygen. The kinetics was still increased in model media containing (+)-catechin, while no color was detected when normal-phase HPLC-fractionated dimers or trimers were investigated. (+)-Catechin emerged as the precursor of less polar products, characterized by a yellow-brown color. MS/MS enabled us to identify these products as issued from the oxidation and intramolecular additions of dehydrodicatechin B4. Similar structures were found in aged beers spiked with (+)-catechin. Beer storage in the absence of oxygen and at low temperature is recommended so as to minimize the synthesis of such pigments.     

How low pH can intensify beta-damascenone and dimethyl trisulfide production through beer aging

Flavor quality is of major importance to the consumer, but the flavor characteristics of beer appear to deteriorate greatly with time, at a rate depending on the composition of the beer and its storage conditions (notably pH). Prior to identifying the influence of pH on the development of the most intense staling flavors found in aged lager beers, the corresponding key flavor compounds were determined by aroma extract dilution analysis. In addition to trans-2-nonenal, beta-damascenone seems at least as important in the flavor of aged beer. Ethyl butyrate, dimethyl trisulfide, 2-acetylpyrazine, 3-(methylthio)propionaldehyde, 2-methoxypyrazine, maltol, gamma-nonalactone, and ethyl cinnamate are also relevant to the sensory profile of aged beer. Upon aging, a beer having a higher pH produces less beta-damascenone, because acid-catalyzed glycoside hydrolysis is decreased. On the other hand, it produces more 3-(methylthio)propionaldehyde, owing to Strecker degradation of methionine. Raising the beer pH additionally causes the release of 3-(methylthio)propionaldehyde from sulfitic adducts. These adducts, more stable at a lower pH, protect the aldehyde against premature oxidation to 3-(methylthio)propionic acid, thus making it available for dimethyl trisulfide formation during aging.     

Correlation of malt quality parameters and beer flavor stability: multivariate analysis

Malt is known to have an impact on beer flavor stability mainly due to the presence of antioxidants. In this study, five barley varieties were malted at industrial and micro scale, and quality parameters of the resulting malts were measured (diastatic power, friability, beta-glucan content, antiradical power, reducing power, lipoxygenase activity, and nonenal potential) and correlated with the sensory data obtained for the corresponding fresh and forced aged beers. A statistical strategy using multiple linear regressions was applied to explore relationships between the malt chemical parameters and beer sensory data, showing antiradical power as the major contribution of malt to beer flavor stability. Additionally, the measured antiradical power, which is well correlated with the polyphenolic content, was found to be very similar for malt and barley, emphasizing the key role of barley endogenous polyphenols.     

Determination of free phenolic acids in wort and beer by coulometric array detection

The simultaneous determination of 19 phenolic compounds was performed directly in wort and beer by a combination of reverse-phase high-performance liquid chromatography coupled with coulometric array detection. Chromatographic separation was achieved with an appropriate gradient of flow and a binary solvent based on phosphate buffer, methanol, and acetonitrile in a 45-min run. Eight serial coulometric detectors were used for on-line generation of voltammetric data to resolve coeluting compounds. The method was reliable and sensitive, the regression coefficient of standard calibration curves is 0.972 < or = r < or = 1.000, and the standard deviation value ranges from 0.010 to 0.129 mg/L for wort and from 0.002 to 0.332 mg/L for beer. The mean concentrations of phenolic acids were 22.1 and 33.8 mg/L, respectively, in worts and beers produced in Italy. These amounts represent 5 and 10% of the non-tannic, non-flavonoid phenols in wort and beer, respectively.     

Investigation of the beta-damascenone level in fresh and aged commercial beers

This study investigated the increase of beta-damascenone content during aging in a variety of commercial Belgian beers. Quantities detected in fresh beers were generally low (from 6 ng/g to 25 ng/g). After 5 days at 40 degrees C, the level increased (to as much as 210 ng/g) in most of the beers studied, according to the type of beer. Further experiments showed that wort initially contains large quantities of beta-damascenone (450 ng/g), but that degradation of the compound during fermentation accounts for the low concentrations observed in fresh beers. Production during beer aging can be partially explained by acid hydrolysis of glycosides.     

Inhibition of induced DNA oxidative damage by beers: correlation with the content of polyphenols and melanoidins

Beers are a source of dietary flavonoids; however, there exist differences in composition, alcohol concentration, and beneficial activities. To characterize these differences, three kinds of lager beer of habitual consumption in Spain, dark, blond, and alcohol-free, were assayed for total phenolic content, antioxidant activity, superoxide and hydroxyl radical scavenging activities, and in vitro inhibitory effect on DNA oxidative damage. Furthermore, their melanoidin content and correlation with antioxidant activity were evaluated. Dark beer contained the highest total phenolic (489 +/- 52 mg/L) and melanoidin (1.49 +/- 0.02 g/L) contents with a 2-fold difference observed when compared to the alcohol-free beer. For the three kinds of beer, the antioxidant activity measured as N,N-dimethyl-p-phenylenediamine dihydrochloride concentration was strongly correlated with the total polyphenol content (R(2) = 0.91102, p < 0.005) and with the melanoidin content (R(2) = 0.7999, p < 0.05). The results support a positive effect of beers on the protection of DNA oxidative damage, by decreasing the deoxyribose degradation, DNA scission (measured by electrophoresis), and inhibition of 8-hydroxydeoxyguanosine (8-OH-dG) formation. Furthermore, a correlation between the total melanoidin content (R(2) = 0.7309, p < 0.01) and inhibition of 8-OH-dG was observed.     

茶叶添加方式对酿造啤酒理化特性及感官品质的影响

在啤酒酿造工艺的煮沸和主发酵阶段分别添加乌龙茶,探讨茶叶不同添加方式对啤酒理化特性、抗氧化能力、啤酒贮存稳定性以及感官特性的影响。结果显示,与未添加茶叶的啤酒产品(对照组CG)相比,在煮沸阶段(煮沸添加组BG)和主发酵阶段(主发酵添加组MG)分别添加0.3g/L的茶叶均提高了酵母发酵速率;添加茶叶的两个茶啤酒产品(煮沸添加组和主发酵添加组)的DPPH自由基清除能力分别提升至82.74%和89.21%、ABTS+自由基清除能力分别提升至41.53%和51.49%、铁离子还原能力分别提升至36.49和43.83 mg/L,且成品茶啤酒贮藏期间的抗老化能力提高;茶啤酒中总酚以及儿茶素(EGC、C、EGCG、EC、GCG、ECG)和咖啡碱(CAF)含量升高,其中,主发酵添加组茶啤酒样品总酚和EGCG含量显著高于对照组和煮沸添加组啤酒(P0.05),分别达到734.40和8.43 mg/L。进一步的感官品评结果显示添加茶叶提高了啤酒产品的茶香气和茶滋味,其中主发酵添加组啤酒的茶香气、茶滋味及酒体协调性最好。由此可知,在啤酒酿造工艺中添加茶叶提高了酵母发酵速率,增强了啤酒中酚类物质含量及其抗氧化和抗老化性能,同时也为啤酒增添了新的茶风味产品。     

Activation of proestrogens from hops (Humulus lupulus L.) by intestinal microbiota; conversion of isoxanthohumol into 8-prenylnaringenin

Hop, an essential ingredient in most beers, contains a number of prenylflavonoids, among which 8-prenylnaringenin (8-PN) would be the most potent phytoestrogen currently known. Although a number of health effects are attributed to these compounds, only a few reports are available about the bioavailability of prenylflavonoids and the transformation potency of the intestinal microbial community. To test these transformations, four fecal samples were incubated with xanthohumol, isoxanthohumol (IX), and 8-PN. Upon incubation with IX, present in strong ales up to 4 mg/L, 36% was converted into 8-PN in one fecal sample and the estrogenic properties of the sample drastically increased. In an experiment with 12 fecal cultures, this conversion was observed in one-third of the samples, indicating the importance of interindividual variability in the intestinal microbial community. Eubacterium limosum was identified to be capable of this conversion (O-demethylation) of IX into 8-PN, and after strain selection, a conversion efficiency of 90% was achieved. Finally, strain supplementation to a nonconverting fecal sample led to rapid and high 8-PN production at only 1% (v/v) addition. Up to now, the concentration of 8-PN in beer was considered too low to affect human health. However, these results show that the activity of the intestinal microbial community could more than 10-fold increase the exposure concentration. Because prenylflavonoids are present in many beers with IX being the major constituent, the results raise the question whether moderate beer consumption might contribute to increased in vivo levels of 8-PN and even influence human health.     

Antioxidant compounds and antioxidant activity in "early potatoes"

The antioxidant content and the antioxidant capacity of both hydrophilic and lipophilic antioxidant extracts from four "early potato" cultivars, grown in two different locations (Racale and Monteroni), were examined. There was a considerable variation in carotenoid content and weak differences in the ascorbic acid concentration of the examined cultivars of "early potato" and between the harvested locations. An increase in both methanol/water (8:2 v/v) and phosphate buffer soluble (PBS) free phenols (70%) and bound phenols (28%) in the extracts from the cultivars grown at Racale site was found and discussed. Examination of individual phenols revealed that chlorogenic acid and catechin were the major phenols present in potato tuber extracts; a moderate amount of caffeic acid and ferulic acid was also detected. The total equivalent antioxidant capacity (TEAC) was higher in the Racale extracts and a highly positive linear relationship ( R (2) = 0.8193) between TEAC values and total phenolic content was observed. The oxyradical scavenging capacity (TOSC) of methanol/water and PBS extracts of peel and whole potatoes against the reactive oxygen species (ROS) peroxyl radicals, peroxynitrite, and hydroxyl radicals was also analyzed. A highly significant linear correlation ( R (2) = 0.9613) between total antioxidant capacity (as a sum of peroxyl radicals + peroxynitrite) and total phenol content of methanol/water extracts was established. Moreover, proliferation of human mammalian cancer (MCF-7) cells was significantly inhibited in a dose-dependent manner after exposure to potato extracts. These data can be useful for "early potato" tuber characterization and suggest that the "early potato" has a potential as a dietary source of antioxidants.