Exploring the composition and potential uses of four hops varieties through different extraction techniques.

Hydrophilic, lipophilic extracts and essential oil of four hops varieties from Slovenia were examined in this study. Lipophilic extracts were obtained by supercritical extraction (SFE), while for hydrophilic extracts ultrasound and microwave extraction were employed. Essential oils were isolated by the hydrodistillation process. The lipophilic composition of essential oils and lipophilic extracts was determined by GC-MS analysis. Monoterpenes and sesquiterpene hydrocarbons were the most abundant class of compounds in oils (62.27-79.65 %), with myrcene being the most abundant constituent. Limonene and trans-caryophyllene were two terpenes determined in all essential oils while only trans-caryophyllene was detected in SFE samples. Antioxidant, antimicrobial, and cytotoxic activity, determined by applying in vitro assays, was more influenced by extraction technique than by varieties. Molecular docking was carried out to gain insight into the potential cancer protein targets BCL-2 and MMP9, whereby humulene epoxide II displayed good binding configuration within the cavities of the two proteins.

Corrigendum to: Impact of Successive Exploitation of a Saccharomyces pastorianus Starter Culture on Saccharide Uptake Dynamics from Wort (Published: Food Technol. Biotechnol. 59 (1) 16-23 (2021) https://doi.org/10.17113/ftb.59.01.21.6899).

[This corrects the article DOI: 10.17113/ftb.59.01.21.6899.].

Extraction of Polyphenols from Slovenian Hop (Humulus lupulus L.) Aurora Variety Using Deep Eutectic Solvents: Choice of the Extraction Method vs. Structure of the Solvent.

Polyphenols from Slovenian hops (Humulus lupulus L.) of the Aurora variety were extracted by different methods and using classical solvents and several deep eutectic solvents (DES) based on choline chloride as the hydrogen bond acceptor component. The obtained extract solutions were analyzed by HPLC for the content of extracted α- and ß-acids and extracted xanthohumol. It was found that choline chloride:phenol DES concentrated aqueous solution had an extraction efficiency close to that of diethyl ether, which is considered one of the best classical extraction solvents for polyphenols from hops. The comparison of the extraction efficiency with other choline chloride-based DESs showed that the chemical similarity of the phenol ring in the solvent DES with the polyphenols in hops may be crucial for a highly efficient extraction with choline chloride:phenol DES. On the other hand, the choice of extraction method and the viscosity of the solvents tested seem to play only a minor role in this respect. As far as we know, this is the first study to attempt to relate extraction efficiency in the extraction of hydrophobic solutes to the compressibility of the DES extractants, the latter of which may be correlated with the extent of hydrophobic hydration around the DES components. In addition, using the heating and stirring method for the preparation of choline chloride-based DES concentrated aqueous solutions we found no support for the occurrence of water in two different roles (in the structural and in the dilution role) in these solvents.

The Stability of Hop (Humulus lupulus L.) Resins during Long-Period Storage.

The stability of alpha-acids, beta-acids and hop storage index (HSI) values under different conditions (aerobic/anaerobic, 4 °C/room temperature) was studied in a two-year trial. Six different varieties (Celeia, Aurora, Bobek, Styrian Gold, Savinjski Golding and Styrian Wolf) were used in the form of cones and pellets. Alpha- and beta-acids were determined by HPLC and HSI by spectrophotometry. Anaerobic conditions at 4 °C were best for alpha-acids, beta-acids and HSI values; however, 10-35% of the alpha-acids were still lost after two years. The decline was greater (63-99%) under aerobic conditions and at room temperature. Alpha-/beta-acid ratios increased in hop cones and decreased in hop pellets, whereas HSI values increased in all storage conditions. Overall, the performance was better for pellets than for hop cones. Storage conditions, storage form and hop variety had significant effects on the stability of hop resins.

The Impact of Hop Freshness on Kettle-Hopped Beers.

Hops are an indispensable ingredient in beer, and the differences in their chemical composition impart the various tastes and aromas associated with different beers. However, during storage, hops undergo changes in their chemical composition. Here, the changes in aroma and bitterness of kettle-hopped beers were evaluated in an experiment conducted on three different hop varieties (Aurora, Celeia and Styrian Wolf) with five different hop storage index (HSI) values (0.3-0.7). Hops were added to boiling wort for 5, 45 and 90 min. Alpha-acids, iso-alpha-acids, humulinones, bitterness units and hop aroma compounds in the samples were chemically analysed. All samples also underwent sensorial analysis. The old hops were not problematic in terms of bitterness or early hopping time. However, later additions of old hops reduced the quality and intensity of the hop aroma. The limit value for use without negative consequences for kettle hopping was set at HSI 0.5 for Aurora and Celeia and HSI 0.6 for Styrian Wolf.

Changes in Hop (Humulus lupulus L.) Oil Content and Composition during Long-Term Storage under Different Conditions.

Hop essential oil, in addition to alpha-acids, is one of the most valuable parameters for brewers, since it is responsible for beer aroma. The hop oil content and hop oil chemical composition deliver various aromas to beer. During storage, the hop chemical composition undergoes many physical and chemical changes that impact its quality. The main purpose of our study was to evaluate the changes occurring in hop oil content and its chemical composition during two years of storage under four different conditions (anaerobic, aerobic, cold room (4 °C), and room temperature) in the form of cones and pellets, supplied by local suppliers in Slovenia. Hop oil content and composition were determined by steam distillation and GC-FID. The greatest decrease in hop essential oil content occurred when the hops were stored under aerobic conditions and at room temperature. Monitoring of eight hop oil compounds under various conditions revealed different behaviors of the changes. The best storage conditions were anaerobic at low temperatures. Oxygen resistance was lower for pellets than for hop cones, whereas high temperature was more devastating for hop cones. In addition to the storage conditions, the hop variety and form were important factors regulating the extent of changes in hop oil content and chemical composition.

Impact of Hop Freshness on Dry Hopped Beer Quality.

The hop plant is seasonal, but beer production continues throughout the whole year. The quality of hops begins to decrease immediately after harvesting; therefore, maintaining the highest possible quality is important. A good indicator of hop freshness is the hop storage index (HSI). In this study, three different varieties of hops with five different HSI values, from 0.3 to 0.7, were used for brewing with the dry hopping technique. The main goal was to evaluate the impact of the HSI value on beer quality in terms of hop aroma and bitterness. Alpha acids, iso-alpha acids, humulinones, bitterness units and hop aroma compounds were chemically analysed. Sensorial analysis was also conducted on all samples. Decreases in the intensity and quality of hop aroma were detected with increasing HSI. The quality of bitterness was also reduced. High HSI also led to undesirable gushing. Beers brewed with hops with HSI values greater than 0.4 had deviations in aroma and bitterness when compared with beers brewed with fresh hops.

Antimicrobial Properties of Different Hop (Humulus lupulus) Genotypes.

The antimicrobial activity of hop extracts obtained from different hop genotypes were investigated against Staphylococcus aureus and Lactobacillus acidophilus. In this study the pure xanthohumol, purified ß-acids rich fraction, as well as α-acids with ß-acids rich fraction were used to test antimicrobial activity against Staphylococcus aureus and Lactobacillus acidophilus; whereby, the antimicrobial activity of different hop extracts against Lactobacillus acidophilus was studied for the first time. Microbial susceptibility to purified hydroacetonic extracts from different hop varieties was investigated by the broth microdilution assay to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The hop hydroacetonic extracts were more effective against Staphylococcus aureus than against Lactobacillus acidophilus. Strong inverse correlations of MIC and MBC values were obtained with xanthohumol, cohumulone, n+adhumulone, colupulone and n+adlupulone contents, suggesting that the identified chemical hop compounds are directly responsible for antimicrobial effects. Moreover, the effect of the growth medium strength on the MIC values of hop extracts against Staphylococcus aureus was systematically investigated for the first time. The current study also reveals the effect of different hop extracts on Staphylococcus aureus, which responds to their presence by lag phase extension and generation time prolongation.

Impact of Successive Exploitation of a Saccharomyces pastorianus Starter Culture on Saccharide Uptake Dynamics from Wort.

RESEARCH BACKGROUND: The production of lager beer includes successive repitchings of a single Saccharomyces pastorianus starter culture. During the beer production process, the yeast is exposed to several stress factors which could affect the fermentation performance. An incomplete fermentation represents a waste of fermentable extract and leads to a beer with higher carbohydrate levels, which could result in a beer with an atypical flavour profile. The aim of the present study is to determine the impact of successive exploitation of a single S. pastorianus starter culture on the wort saccharide uptake dynamics. EXPERIMENTAL APPROACH: The fermentation was monitored during the production of twelve batches of beer, where the starter yeast culture was reused twelve times without any further treatment. The following beer production steps were monitored: wort production, yeast starter culture propagation, primary fermentation, secondary fermentation and the final product. The work was conducted on an industrial scale employing standard process conditions. RESULTS AND CONCLUSIONS: Monitoring of the starter culture viability during successive fermentations indicated no reduction in the viability and vitality of the yeast culture. Monitoring of the fermentable wort saccharide concentrations (glucose, fructose, disaccharides and trisaccharides) revealed a correlation between an improvement in saccharide utilisation and starter culture age. Saccharide uptake efficacy proportionally matched the repitching frequency. Successive exploitation of S. pastorianus starter culture has a positive impact on the dynamics of saccharide utilisation from classical hopped wort and the young beer. Furthermore, the final lager beer contains no residues of fermentable saccharides that could affect the overall quality and flavour profile. NOVELTY AND SCIENTIFIC CONTRIBUTION: Results showed the impact of twelve successive wort fermentations on the dynamics of saccharides uptake that gives brewers important information. The added value of the experiment is all the work done on the industrial scale, with control of all processes and usage of exactly the same raw materials. This study contains usable technological data on the behaviour of saccharides during brewing on the industrial scale, which is not yet found in the literature.

Metabolomic Analysis of Cannabinoid and Essential Oil Profiles in Different Hemp (Cannabis sativa L.) Phenotypes.

Hemp (Cannabis sativa L.) cannabinoids and terpenoids have therapeutic effects on human and animal health. Cannabis plants can often have a relatively high heterogeneity, which leads to different phenotypes that have different chemical profiles despite being from the same variety. Little information exists about cannabinoid and terpenoid profiles in different hemp phenotypes within the same variety. For this study, 11 phenotypes from three different varieties ("Carmagnola" selected (CS), "Tiborszallasi" (TS), and "Finola" selection (FS)) were analyzed. The components of essential oil (29) were analyzed using gas chromatography with flame ionization detection (GC/FID), and 10 different cannabinoids of each phenotype were determined using high-performance liquid chromatography (HPLC). Principal component analysis (PCA) and analysis of variance (ANOVA) showed that according to the components of essential oil, FS and TS plants were more uniform than CS plants, where there were great differences between CI and CII phenotypes. The content of cannabinoid CBD-A was the highest in all four FS phenotypes. By comparing cannabinoid profiles, FS was clearly separated from TS and CS, while these two varieties were not clearly distinguishable. Phenotypes TV and CI had the highest total content of Δ-9-THC, while all phenotypes of FS had the highest total content of CBD. The highest total content of CBG was determined in phenotype CI. Obtained results are useful for the development of new supplementary ingredients, for different pharmacy treatments, and for further breeding purposes.

Dynamics of Isomerization of Hop Alpha-Acids and Transition of Hop Essential Oil Components in Beer.

Hops' unique composition of essential oil components and bitter resins are crucial for beer aroma, which is important to consumers' acceptance of beer. In this experiment the same wort was divided into four portions and each was hopped differently. To determine the dynamics of isomerization rates the concentrations of alpha- and iso-alpha-acids were continuously measured. Measurements of hop essential oil components were performed during each process to understand the dynamics of the transition into beer. The maximum isomerization yield of alpha-acids (18.1%) was achieved after 100 min. Longer boiling increased the reduction of iso-alpha-acids, as well as essential oil components. Dry hopping also affected not only on beer aroma but also on beer bitterness.

The determination of the geographic origins of hops (Humulus lupulus L.) by multi-elemental fingerprinting.

For brewers, it is important to know the geographic origins of the hop plants (Humulus lupulus L.) used in their brewing processes since the contents and compositions of the bitter resins and essential oils in them depend on the environmental conditions during their growth. In this study, the multi-elemental profiles of hop plants from each of the world's main growing regions were determined by non-destructive energy dispersive X-ray fluorescence spectrometry (EDXRF). Understanding the multi-element profiles of the plants could be a useful tool in determining the geographic origins of produced hop plants.

Determination of the Geographical and Botanical Origin of Hops (Humulus lupulus L.) Using Stable Isotopes of C, N, and S.

A need exists for a reliable method to determine the geographical and botanical origin of hops. For this study, three sets of samples were collected: the first set comprised 5 German samples; the second set comprised samples of hops from 10 of the world's major hop-growing regions; and the third comprised the 4 main Slovenian regions. The samples were analyzed using isotope ratio mass spectrometry (IRMS) to obtain δ13C, δ15N, and δ34S values. The δ15N (2.2 ‰ to 8.4 ‰) and δ34S (0.7 ‰ to 12.3 ‰) values were the most discriminating parameters for classifying hop according to geographical origin. ANOVA showed distinct groupings for 8 out of the 10 hop-growing regions. Although it was not possible to distinguish the geographical origin of hops based on δ13C (-28.9 ‰ to -24.7 ‰), in the case of botanical origin, δ13C values proved to be the most discriminative albeit with limited success.

Comparison of sensory and chemical evaluation of lager beer aroma by gas chromatography and gas chromatography/mass spectrometry.

BACKGROUND: Although the evaluation of beer is conducted by sensory experts, we cannot neglect the influence of human factors and subjectivity. This problem could be solved by the chemical analysis of the volatile part of beer aroma and, from this, we can build a database for the construction of a model that classifies samples in a comparable manner to sensory assessment. RESULTS: Twenty-two batches of the same beer brand were assessed by sensory evaluation and described chemically in terms of the contents of alcohols and esters (n = 9), hop essential oil compounds (n = 15) using gas chromatography (GC) and other aroma volatiles (n = 33) as analysed by head space solid-phase microextraction (SPME)-GC/mass spectrometry. The best match of 91% with respect to sample classification on the basis of chemical analyses to sensory scores was achieved with a dataset of results from headspace-SPME and analyses of higher alcohols and esters by regularized discriminant analysis. CONCLUSION: The results of the present study show that deviations in beer aroma are not a consequence of a permanent repeatable error in brewing process, nor are they influenced by raw materials, but, instead, they are a consequence of alcoholic fermentation. Sensory analysis could be replaced with chemical/statistical analysis on an appropriate data set and for a distinct beer brand. The good results achieved confirm our approach; however, for different beer brands or types, this method should be optimised. © 2017 Society of Chemical Industry.