Effect of Wort Boiling System and Hopping Regime on Wort and Beer Stale-Flavor Aldehydes.

The main factor responsible for the sensory aging of beer is the increase in off-flavor aldehydes during beer storage. In pilot brews (200 L) of pale lager beer with different hopping regimes and wort boiling systems, 15 carbonyls were monitored using the GC-MS method. Factor analysis revealed several groups of aldehydes with similar behavior during wort boiling. The concentration of most of them decreased with atmospheric wort boiling and increased with the time and energy-saving pressurized boiling system. Wort clarification was a critical step because of the increase in carbonyl concentration, with the level of most carbonyls being higher in the final wort compared to sweet wort. The hopping regimes only affected the level of 3-methylbutan-2-one in the wort. The concentration of carbonyls decreased significantly (30-90%) during fermentation, except for trans-2-butenal, which increased by 59% on average, likely due to the release from imine complex. The concentration of free aldehydes in the fresh beers was similar for all variants used, but the pressurized wort boiling system could result in lower sensory stability of the beer due to the release of aldehydes from inactive complexes formed during fermentation. This aspect requires further investigation.

Inhibitory effect of hop fractions against Gram-positive multi-resistant bacteria. A pilot study.

AIM: Our research focused on the antimicrobial effects of purified hop (Humulus lupulus L.) fractions including α-bitter acids (humulones), ß-bitter acids (lupulones) and xanthohumol, and a commercial CO2 hop extract of bitter acids against reference and multi-resistant strains of Gram-positive and Gram-negative bacteria and against selected yeast strains. METHODS: In vitro testing of antimicrobial activity was performed according to standard testing protocols (EUCAST). The effects of hop extracts on bacterial/yeast strains at concentrations below MICs were also determined and the antimicrobial potential of hop extracts was compared with selected antibiotics using optical density measurement. RESULTS: The fractions were effective not only against reference strains of Gram-positive bacteria but, more importantly, against their methicillin- and vancomycin-resistant variants. No antimicrobial effect was detected against Gram-negative bacterial strains. Among the tested substances, xanthohumol was identified as the hop fraction with the most potent antimicrobial properties. It was also found that hop substances exerted their antimicrobial effects at concentrations considerably lower than the determined MICs, with the strongest effect in case of α-bitter acids in enterococci. CONCLUSION: The search for and research of new compounds with antimicrobial properties represents a possible solution to the current global problem of bacterial resistance. Our data suggest a desirable activity of hop fractions against some multi-resistant bacterial strains. Thus, hops might find use as a source of potential antimicrobial agents applicable in both human and veterinary medicine.

Strong antimicrobial activity of xanthohumol and other derivatives from hops (Humulus lupulus L.) on gut anaerobic bacteria.

Anaerobic bacteria, such as Bacteroides fragilis or Clostridium perfringens, are part of indigenous human flora. However, Clostridium difficile represents also an important causative agent of nosocomial infectious antibiotic-associated diarrhoea. Treatment of C. difficile infection is problematic, making it imperative to search for new compounds with antimicrobial properties. Hops (Humulus lupulus L.) contain substances with antibacterial properties. We tested antimicrobial activity of purified hop constituents humulone, lupulone and xanthohumol against anaerobic bacteria. The antimicrobial activity was established against B. fragilis, C. perfringens and C. difficile strains according to standard testing protocols (CLSI, EUCAST), and the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBC) were calculated. All C. difficile strains were toxigenic and clinically relevant, as they were isolated from patients with diarrhoea. Strongest antimicrobial effects were observed with xanthohumol showing MIC and MBC values of 15-107 µg/mL, which are close to those of conventional antibiotics in the strains of bacteria with increased resistance. Slightly higher MIC and MBC values were obtained with lupulone followed by higher values of humulone. Our study, thus, shows a potential of purified hop compounds, especially xanthohumol, as alternatives for treatment of infections caused by select anaerobic bacteria, namely nosocomial diarrhoea caused by resistant strains.

A novel approach for identification of biologically active phenolic compounds in complex matrices using hybrid quadrupole-orbitrap mass spectrometer: A promising tool for testing antimicrobial activity of hops.

The phenolic compounds, secondary metabolites of hops represent a large family of compounds that could be subsequently divided into smaller groups based on the similarities between their chemical structures. The antibacterial, antifungal and antiviral properties of hops are well known, but there is a lack of information about antimicrobial activities of individual hop compounds. This study was carried out with an objective to identify compounds present in hops that have potential antibacterial activity. In the first stage of experiment, the active compounds with potential anti-microbial activity had to be extracted from hop cones. Therefore, minced hop cones were applied on solid growth medium inoculated with Staphylococcus aureus. The active substances that migrated into the medium created an inhibition zone. In the second stage of experiment, the inhibition zones were cut out from Petri dishes, active compounds were extracted from these zones and consequently analyzed using LC-HRMS. These complex assays were developed and optimized. The data were acquired by using a quadrupole-orbitrap hybrid mass spectrometer by targeted-MS2 experiment in both ionization modes. The MS method has been developed as a screening method with a subsequent fragmentation of compound of interest on the base of inclusion mass list. The unknown compounds extracted from inhibition zones have been identified either by searching against a database or their structure has been elucidated on the basis of their fragmentation spectra. On the basis of this experiment the list of active compounds with potential anti-microbial activities was enhanced.

Changes in polyphenol compounds and barley laccase expression during the malting process.

BACKGROUND: Polyphenols and phenolic acid are able to slow down or prevent oxidation processes and are therefore thought to have important effects in malting and brewing. Laccase catalyses the oxidation of a wide variety of substrates, including polyphenols. The aim of this paper was to determine the changes in polyphenol compounds and the relative expression of the HvLac1 gene during malting. RESULTS: The dominant phenolic acid was ferulic acid. The amount of ferulic acid increased, whereas the amount of vanillic acid decreased during malting. The highest levels of expression of the HvLac1 gene were observed during the third air rest period in varieties with the 'Haruna Nijo' (HN) allele, as recommended for the production of beer with the protected geographical indication (PGI) 'Ceske pivo' (Czech beer), whereas the highest expression was observed in the first day of germination in varieties with the 'Morex' (M) allele. However, the profiles of HvLac1 gene expression in varieties with alternative alleles during malting were similar, and the level of polyphenol compounds throughout malting was different. CONCLUSION: The polyphenol contents in barley increased several-fold during malting, and the degree of increase differed with variety. The expression of HvLac1 transcript was similar in every barley variety.

Qualitative determination of ß-acids and their transformation products in beer and hop using HR/AM-LC-MS/MS.

Hops represent an important source of ß-acids with antimicrobial and sensory properties. Transformation products of ß-acids formed during their oxidation, mainly hulupones, have been shown to have an interesting kind of bitterness. Their structures were recently elucidated using LC-TOFMS and 1D/2D NMR in solution after thermal treatment of the hop ß-acids. This study demonstrates the advantages of MS detection with high resolution and accurate mass measurements. The structure of transformation products in an experimental solution of oxidized ß-acids was elucidated using a newly developed method by hybrid quadrupole-Orbitrap MS. In addition to already known structures, two new ones were identified and named epoxycohulupone and epoxyhulupone. The method was verified on real samples; the profiles of these products in Sládek hops harvested in 2008 and 2012 and in corresponding beers were compared. For this purpose, a new QuEChERS assay was used for the preparation of beer samples.