Bioactive Compounds Obtained from Polish “Marynka” Hop Variety Using Efficient Two-Step Supercritical Fluid Extraction and Comparison of Their Antibacterial,Cytotoxic, and Anti-Proliferative Activities In Vitro

Given the health-beneficial properties of compounds from hop, there is still a growing trend towards developing successful extraction methods with the highest yield and also receiving the products with high added value. The aim of this study was to develop efficient extraction method for isolation of bioactive compounds from the Polish “Marynka” hop variety. The modified two-step supercritical fluid extraction allowed to obtain two hop samples, namely crude extract (E1), composed of α-acids, β-acids, and terpene derivatives, as well as pure xanthohumol with higher yield than that of other available methods. The post-extraction residues (R1) were re-extracted in order to obtain extract E2 enriched in xanthohumol. Then, both samples were subjected to investigation of their antibacterial (anti-acne, anti-caries), cytotoxic, and anti-proliferative activities in vitro. It was demonstrated that extract (E1) possessed more beneficial biological properties than xanthohumol. It exhibited not only better antibacterial activity against Gram-positive bacteria strains (MIC, MBC) but also possessed a higher synergistic effect with commercial antibiotics when compared to xanthohumol. Moreover, cell culture experiments revealed that crude extract neither inhibited viability nor divisions of normal skin fibroblasts as strongly as xanthohumol. In turn, calculated selectivity indexes showed that the crude extract had from slightly to significantly better selective anti-proliferative activity towards cancer cells in comparison with xanthohumol.     

Analysis of xanthohumol and isoxanthohumol in different hop products by liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry

An analytical methodology based on the sample extraction with methanol/formic acid by ultra-sonication and subsequent analysis by high-performance liquid chromatography with diode array detection is proposed for the determination of xanthohumol (XN) and isoxanthohumol (IXN) in different hop products. The identity of the compounds was confirmed by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry in positive ion mode. The performance of the method was assessed by the evaluation of parameters such as absolute recovery, repeatability, linearity and limits of detection and quantitation. This methodology was applied to investigate the impact of the extraction process of the hop products on the amount of xanthohumol and isoxanthohumol. The ethanolic extract revealed to be the hop product richest in xanthohumol (3.75+/-0.05 g/100 g) relatively to the pellets (0.62+/-0.01 g/100 g) and supercritical CO2 extract (0.089+/-0.001 g/100 g).     

高效液相色谱法测定酒花α-酸含量

介绍反相液相色谱测定酒花和酒花浸膏中的α-酸含量的方法,反相色谱分离α-酸,外标法定量。方法具有样品处理简单、快速、定量准确和重复的特点。酒花和酒花浸膏的α-酸测定相对偏差分别为2.02％和1.78％。     

Determination of xanthohumol in hops (Humulus lupulus L.) by nonaqueous CE

Xanthohumol (XN) is a prenylated chalcone with antimutagenic and anticancer activity from hops. A nonaqueous reverse polarity capillary electrophoretic method for the determination of XN in hop extract was developed and validated. The optimal parameters were a 64.5 cm long fused-silica capillary with 50 microm id at 25 degrees C; 30 kV negative voltage (anode at detector side of the capillary); nonaqueous buffer with 75 mM NaOH and 50 mM boric acid in methanol; hydrodynamical injection with 10 mbar for 40 s; and detection at 440 nm. XN, isoxanthohumol (IX), colupulone, adlupulone, and n-lupulone were well resolved on the electropherogram. The LOD for XN was 0.05 mg/L and RSD for peak area was below 3%. The amount of XN in different samples of hop pellets varied from 0.14 to 0.42%.     

Use of hop cone extract obtained under supercritical CO2 conditions for producing antibacterial all-purpose cleaners

This paper attempts to demonstrate empirically the possibility of using a hydrophobic extract of hop cone obtained under the conditions of supercritical carbon dioxide to produce all-purpose cleaner with antimicrobial activity and low irritant potential. A series of prototypes of products varied by the extract concentration was developed. The formulations were assessed for physicochemical properties: particle size, turbidity, and color. The results indicate that the hydrophobic extracts of hop cone can be a valuable component of chemical products intended for washing, having a broad spectrum of action and contributing to improvement of the product’s safety.     

Direct characterization of bitter acids in a crude hop extract by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

The applicability of on-line coupling of reversed-phase high-performance liquid chromatography to atmospheric pressure ionization tandem mass spectrometry for the separation and characterization of hop acids mixture from the crude extract of Humulus lupulus was investigated. The solvent system consisting of acetonitrile-aqueous formic acid was used to give proper separation of the six main hop bitter acids within 30 min. Further structural information about the components was acquired by collision-induced dissociation (CID). On the basis of analyses of the fragmentation patterns of the major alpha- and beta-bitter acids respectively, identification of the minor ones was performed using selected reaction monitoring (SRM) with a group of qualitatively relevant selected precursor-product ion transitions for each bitter acid in a single high performance liquid chromatography (HPLC) run. Using this technique, six minor hop acids, including "adprelupulone" observed for the first time in natural resources, were detected along with the six major acids. This hyphenated techniques provides potency for rapid qualitative determination of analogs and homologs in mixtures.     

Comparison of four extraction methods for analysis of volatile hop‐derived aroma compounds in beer

The volatile organic compound profile in beer is derived from hops, malt, yeast, and interactions between the ingredients, making it very diverse and complex. Due to the range and diversity of the volatile organic compounds present, the choice of the extraction method is extremely important for optimal sensitivity and selectivity. This study compared four extraction methods for hop‐derived compounds in beer late hopped with Nelson Sauvin. Extraction capacity and variation were compared for headspace solid‐phase micro extraction, stir bar sorptive extraction, headspace sorptive extraction, and solvent‐assisted flavor evaporation. Generally, stir bar sorptive extraction was better suited for acids, headspace sorptive extraction for esters and aldehydes, while headspace solid‐phase microextraction was less sensitive overall, extracting 40% fewer compounds. Solvent‐assisted flavor evaporation with dichloromethane was not suitable for the extraction of hop‐derived volatile organic compounds in beer, as the profile was strongly skewed towards alcohols and acids. Overall, headspace sorptive extraction is found to be best suited, closely followed by stir bar sorptive extraction.     

三角形法和四面体法优化选择毛细管区带电泳背景电解质

建立了两种高效、快速的毛细管区带电泳背景电解质(BGE)的优化方法三角形优化法和四面体优化法。以色谱指纹图谱指数F和色谱指纹图谱相对指数Fr作为评价毛细管电泳分析系统的目标函数,以雪莲药材水提取液为样品,考察一定浓度的硼砂、硼酸、磷酸氢二钠和磷酸二氢钠溶液按三角形优化法和四面体优化法构成背景电解质时对样品的分离情况,通过添加有机改性剂和调节pH进行再优化。用三角形法优化出以50 mmol/L硼砂-含3%乙腈的150 mmol/L磷酸二氢钠(体积比为1∶1)作为BGE时分离效果最佳,用四面体法优化出以50 mmol/L硼砂-150 mmol/L磷酸二氢钠-200 mmol/L硼酸(体积比为1∶1∶2,用0.1 mol/L氢氧化钠调pH 8.55)作为BGE时分离效果最佳,分别获得28个和25个电泳峰。所建立的方法操作简捷,适用于中药材水提取液或醇提取液的毛细管区带电泳BGE的选择。     

Comparison of different extraction methods: steam distillation, simultaneous distillation and extraction and headspace co-distillation, used for the analysis of the volatile components in aged flue-cured tobacco leaves

Steam distillation (SD), simultaneous distillation and extraction (SDE) and headspace co-distillation (HCD) were compared here for their effectiveness in the extraction of volatile compounds from tobacco. The different grades of aged flue-cured tobacco leaves extracted by the three methods respectively were analyzed using GC-MS. Mass spectra or authentic compounds were used to identify around 408 components in various volatile fractions. On the one hand, the qualitative comparison showed that more compounds were detected in HCD extract (391 components) than in SDE extract (377 components), and the approximately quantitative analysis showed that the total amount of volatile components in SDE extract (445.48 microg/g) was much more than that in HCD extract (315.72 microg/g). But on the other hand, HCD was the most efficient for nearly all the highly volatile compounds among the three methods. As to low-volatile compounds such as lactones, long chain aldehydes, ketones, alcohols, and esters, more was detected in SDE extract than in HCD extract. The SD method (322 components, total amount 228.42 microg/g) was the lowest sensitive to all compounds except semi-volatile fatty acids among the three methods.     

Chromatographic methods for the determination of mycotoxins in food products

Chromatographic methods for the determination of mycotoxins from different classes in food products of plant and animal origins are surveyed. The procedures of sample preparation and extract purification and the use of various chromatographic analysis techniques are considered.     

Analysis of beer iso-α-acids by micellar electrokinetic chromatography and multi-wavelength UV detection

The identification of beer bitter acids in a pre-isomerized hop extract by means of their UV-spectra is described. Preliminary data for the analysis of these compounds in beer samples are presented.     

Capillary electrophoresis in association with chemometrics approach for bitterness hop (Humulus lupulus L.) classification

The precursor compounds related to the bitterness of beer are called α‐acids. These compounds are extracted from the hop, which is an important ingredient in the brewing process. These compounds were analyzed by capillary electrophoresis. The electrophoretic method used 160 mmol/L of ammonium carbonate (pH 9) as BGE (background electrolyte), a voltage of +20 kV in a capillary with 50 μm of internal diameter and with a 62.5 cm of total length (54 cm effective). The samples were injected in hydrodynamic mode applying a pressure of 25 mbar for 5 s and the analytes were detected at 230 nm. A hydromethanolic extraction during 3 h was considered as the optimum condition for the sample preparation using MeOH/H₂O 80:20 v/v as the extract solution. From the optimized conditions the electropherograms were evaluated for their use as input for chemometric modeling. Preprocessing investigation for electrophoretic data taking into account the alignment, denoising and baseline correction, and variable selection were considered before the chemometric modeling using principal component analysis (PCA). The electrophoretic data were systematically evaluated to find the optimum conditions to modeling. A PCA analysis for all tests was carried out using different preprocessing methods and, an explained variance higher than 90% was achieved in all of them. The optimized chemometric method worked with aligned and meancentered data. From this approach, a simple and efficient method to classify hop samples with high and low α‐acids content without the use of analytical standards was established from a simple electrophoretic analysis.     

The Influence of High-Intensity Ultrasonication on Properties of Cellulose Produced from the Hop Stems,the Byproduct of the Hop Cones Production

The goal of this work is to evaluate the hop stems, a byproduct of hop cones production, as a potential source of cellulose. Hop stems contain up to 29% of cellulose. The cellulose isolation was conducted through the thermochemical treatment. After high-speed blending, the cellulose was characterized by 67% of crystallinity degree obtained from X-ray diffraction and median diameter of 6.7 nm obtained from atomic force microscopy imaging. The high-intensity ultrasonication (HIUS) was applied to reach further disintegration of cellulose fibers. The longer HIUS treatment resulted in decrease in crystallinity degree even up to 60% and decrease in the fiber diameter up to 4 nm. The Fourier transform infrared spectroscopy (FTIR) spectra showed that HIUS treatment led to changes in intermolecular hydrogen bonds. The stability of cellulose dispersions versus length of HIUS treatment was monitored over 14 days with back dynamic light scattering and laser Doppler electrophoresis methods. Obtained results are evidence that the hop stems are a potential source of cellulose and that it is possible to obtain stable dispersions after HIUS treatment. This was the first time that the properties of hop cellulose have been described so extensively and in detail after the use of HIUS treatment.     

Comparison of green extraction methods with conventional extraction method for extract yield,L-DOPA concentration and antioxidant activity of Mucuna pruriens seed

ABSTRACT

Mucuna pruriens is a plant of Fabaceae family. Seed of M. pruriens is considered as a rich source of levo-3,4 dihydroxyphenylalanine (L-DOPA), a non-protein phenolic amino acid. In the present study, three different extraction methods were compared for extract yield, concentration of bioactive compounds such as total phenol, L-DOPA and antioxidant capacity. Extracts were prepared using water acidified with hydrochloric acid (0.1?N) by conventional method of refluxing as well as two green methods namely ultrasound and microwave assisted solvent extraction. A rapid and qualified high-performance liquid chromatography method was also developed for quantification of L-DOPA in different extracts. Among the three extraction methods, microwave assisted extraction provided the best results for yield and quality of M. pruriens extract in much shorter time in comparison to refluxing method of extraction.     

Antioxidant Capacity and Total Phenol Content in Hop and Malt Commercial Samples

The antioxidant properties of commercial hops and malted barley were evaluated by electroanalytical methods and compared with spectrophotometric methods. Hop group showed better antioxidant capacity and total phenols content (TPC) values than malt group. The highest electrochemical Index (EI) was observed for HCO hop sample. Electrochemical Index (EI) showed a good Pearson correlation with ABTS (2,2′‐azino‐bis (3‐ethylbenzothiazoline‐6‐sulphonic acid)), DPPH (2,2‐diphenyl‐1‐picrylhydrazyl), and TPC, being of 0.85, 0.77 and 0.86 respectively for hop samples. While for the malt samples were 0.96, 0.69 and 0.49, respectively. The samples were compared to each other within each group and statistical differences can be observed, p<0.05. Therefore, the electroanalytical tools showed to be practical and easy to evaluate the antioxidant properties of hop and malt samples. The results may contribute to the selection of raw material for brewing and therefore with the quality of one of beverage one most consumed around the world.     

Determination of methyl tert-butyl ether in gasoline: a comparison of three fast methods based on mass spectrometry

A high-speed quantitative analysis of methyl tert-butyl ether (MTBE) using three different methods with mass spectrometry detection has been performed. The first method is based on fast chromatography and required an analysis time of 5.23 min per sample, although a certain period (6 min) was necessary for the initial measurement conditions to be regained prior to analysing the next sample. The other two are non-separative methods and are based on direct injection and headspace generation. The analysis times were 1.5 and 3.5 min, respectively, although in the latter case an additional period of time was required to extract volatiles from the sample. The analytical characteristics of all three methods are highly satisfactory in terms of linearity, lack of fit, precision and accuracy. The methods were applied to the determination of MTBE in different gasoline samples. The non-separative methods afforded slightly higher concentrations than those found when fast chromatography was used; this is due to the presence of other minor components that contribute to the abundance of the ion at m/z 73, characteristic of MTBE. We propose a correction that removes this error very satisfactorily and allows the same results to be obtained with all three methodologies proposed.     

Clustering analysis of different hop varieties according to their essential oil composition measured by GC/MS

This study describes the analysis of total hops essential oils from 18 cultivated varieties of hops, five of which were bred in Lithuania, and 7 wild hop forms using gas chromatography-mass spectrometry. The study sought to organise the samples of hops into clusters, according to 72 semi-volatile compounds, by applying a well-known method, k-means clustering analysis and to identify the origin of the Lithuanian hop varieties. The bouquet of the hops essential oil was composed of various esters, terpenes, hydrocarbons and ketones. Monoterpenes (mainly β-myrcene), sesquiterpenes (dominated by β-caryophyllene and α-humulene) and oxygenated sesquiterpenes (mainly caryophyllene oxide and humulene epoxide II) were the main compound groups detected in the samples tested. The above compounds, together with a-muurolene, were the only compounds found in all the samples. Qualitative and quantitative differences were observed in the composition of the essential oils of the hop varieties analysed. For successful and statistically significant clustering of the data obtained, expertise and skills in employing chemometric analysis methods are necessary. The result is also highly dependent on the set of samples (representativeness) used for segmentation into groups, the technique for pre-processing the data, the method selected for partitioning the samples according to the similarity measures chosen, etc. To achieve a large and representative data set for clustering analysis from a small number of measurements, numerical simulation was applied using the Monte Carlo method with normal and uniform distributions and several relative standard deviation values. The grouping was performed using the k-means clustering method, employing several optimal number of clusters evaluation techniques (Davies-Bouldin index, distortion function, etc.) and different data pre-processing approaches. The hop samples analysed were separated into 3 and 5 clusters according to the data filtering scenario used. However, the targeted Lithuanian hop varieties were clustered identically in both cases and fell into the same group together with other cultivated hop varieties from Ukraine and Poland.     

Quantification of xanthohumol, isoxanthohumol, 8-prenylnaringenin, and 6-prenylnaringenin in hop extracts and derived capsules using secondary standards

Hop is a well-known and already frequently used estrogenic phytotherapeutic, containing the interesting prenylflavonoids, xanthohumol (XN), isoxanthohumol (IXN), 8- and 6-prenylnaringenin (8-PN and 6-PN). Since the use of secondary standards can form a solution whenever the determination is required of certain components, not commercially available or too expensive, it was decided to develop an accessible HPLC-DAD method for the determination of these prenylflavonoids. The amounts were determined in hop extract and capsules, using quercetin and naringenin as secondary standards. After optimization of the sample preparation and HPLC conditions, the analysis was validated according to the ICH guidelines. The response function of XN, 8-PN, quercetin and naringenin showed a linear relationship. For the determination of XN, a calibration line of at least three concentrations of quercetin has to be constructed. The correction factors for XN (quercetin) and for 8-PN (naringenin) were validated and determined to be 0.583 for XN, and 1.296 for IXN, 8-PN and 6-PN. The intermediate precision was investigated and it could be concluded that the standard deviation of the method was equal considering time and concentration (RSD of 2.5-5%). By means of a recovery experiment, it was proven that the method is accurate (recoveries of 96.1-100.1%). Additionally, by analysing preparations containing hop extracts on the Belgian market, it was shown that the method is suitable for its use, namely the determination of XN, IXN, 8-PN and 6-PN in hop extract and capsules, using quercetin and naringenin as secondary standards.     

Hopping time of a hard disk fluid in a narrow channel

We use Monte Carlo (MC) and molecular dynamics (MD) methods to study the self-diffusion of hard disk fluids, confined within a narrow channel. The channels have a pore radius of Rp, above the passing limit of hard disk diameter (sigma(hd)). We focus on the average time (tau(hop)) needed for a hard disk to hop past a nearest neighbor in the longitudinal direction. This parameter plays a key role in a recent theory of the crossover from single-file diffusion to the bulk limit. For narrow channels near the hopping threshold (Rp=1 in units of sigma(hd)), both MC and MD results for tau(hop) diverge as approximately (Rp-1)(-2). Our results indicate that the scaling law exponent does not appear to be dependent on the differences between the two dynamics. This exponent is consistent with the prediction of an approximate transition state theory.