Beer and beer-based beverage contain lignans.

Lignans are members of a broad group of plant phenols that can positively affect human health. They occur in negligible quantities in processed foodstuffs such as lager beer. The aim of this work was to utilize the high levels of lignans in the knots of spruce trees (Picea abies) to increase the lignans content in beer, without negatively impacting the natural taste and aroma. By means of lignans addition in the forms of spruce knot chips or different extracts made from spruce knots during the wort boiling were produced beer and beer-based beverages with lignans content ranging from 34 to 174 mg/L.

Xanthones Content in Swertia multicaulis D. Don from Nepal.

The medicinal plant Swertia multicaulis D. Don was collected in Rasuwa District (Nepal) and the xanthone content of its ethyl acetate extracts was studied. The total amount of xanthones in S. multicaulis determined by HPLC reaches almost 13 g of xanthones per 1 kg of dry matter. The identification of xanthones in S. multicaulis was achieved by a combination of HPLC, LC⁻MS and LC⁻NMR. The final assignment of the individual chemical structures was provided by NMR, supported by preparative HPLC. In eight chromatographic peaks, four major xanthones were identified­1,3-dihydroxy-5,8-dimethoxyxanthone, 1-hydroxy-3,5,8-trimethoxyxanthone, bellidifolin (1,5,8-tri-hydroxy-3-methoxyxanthone), and decussatin (1-hydroxy-3,7,8-trimethoxyxanthone).

Variability in the Content of Trans-Resveratrol, Trans-ε-Viniferin and R2-Viniferin in Grape Cane of Seven Vitis vinifera L. Varieties during a Three-Year Study.

Grape canes are a waste product from viticulture that show potential as an industrially extractable source of stilbenes, which are valuable for medical and other purposes. In this work, grape canes collected in three consecutive years (2014-2016) at six different places in South Moravia, Czech Republic were extracted, and the contents of trans-resveratrol, trans-ε-viniferin, and r2-viniferin were determined by high-performance liquid chromatography. The study included three blue grape varieties of Vitis vinifera L. (Cabernet Moravia, Blaufränkisch, and Piwi variety Laurot) and four white grape varieties (Chardonnay, Green Veltliner, Piwi variety Hibernal, and Piwi variety Malverina). From the viewpoint of producing extracts with high stilbenes content, the Hibernal variety is clearly the best. The mean amounts of the stilbenes for this variety at all localities and for all three years were 4.99 g/kg for trans-resveratrol, 3.24 g/kg for trans-ε-viniferin, and 1.73 g/kg for r2-viniferin. The influence of vintage, locality, and variety on the amounts of stilbenes was studied using PCA analysis. In contrast to expectations, there was no strong impact of locality on stilbenes content. The differences were varietal for most varieties, regardless of the area of cultivation. Laurot and Hibernal varieties did differ significantly in that respect, however, as they exhibited clear dependence on location.

Larvicidal activity of extracts from Ammi visnaga Linn. (Apiaceae) seeds against Culex quinquefasciatus Say. (Diptera: Culicidae).

Efficacies of the Ammi visnaga seeds extract and a majority of substances on larval Culex quinquefasciatus mortality in various development stages including pupae were studied. The effect of exposure time on larval mortality was also studied. The effect of sublethal concentrations or short exposure times on further larval development and subsequent fecundity in adults were studied as well. Lethal doses of the extract were estimated for the 2nd, 3rd and 4th instar of C. quinquefasciatus (LC50 for 18, 23 and 180 mg L(-1), respectively). The majority of furanochromenes, khellin and visnagin, were identified by analysing the extract. Khellin was significantly more effective compared to visnagin, whose LC50 was estimated at 8, 10 and 41 mg L(-1) for the 2nd, 3rd and 4th instar larvae. Khellin showed very fast efficacy on mortality for the 3rd instar larvae in a concentration of 100 mg L(-1). Fifty percent mortality was determined 30 min after application, a time which was considerably shorter compared to the extract (113 min) or visnagin (169 min). The effect of the application of lethal concentrations on C. quinquefasciatus larval mortality was studied. The least number of adults were hatched after application of the extract and khellin (41.8% and 37.9%, respectively), less than after visnagin application (46.7%) or in the control (94.2%). LC50 application caused lower fecundity in the hatched adults, lower hatchability of the eggs, and also very low natality, more than 77% lower for khellin compared to the control. A short exposure, corresponding to our estimated LT30, caused no significant acute toxicity in the larvae (until 24 h) for the extract or visnagin (4.3% and 11.5%, respectively); however, 18 min of action from khellin caused a 54.3% mortality rate of the larvae within 24 h.

Various extraction methods for obtaining stilbenes from grape cane of Vitis vinifera L.

Grape cane, leaves and grape marc are waste products from viticulture, which can be used to obtain secondary stilbene derivatives with high antioxidant value. The presented work compares several extraction methods: maceration at laboratory temperature, extraction at elevated temperature, fluidized-bed extraction, Soxhlet extraction, microwave-assisted extraction, and accelerated solvent extraction. To obtain trans-resveratrol, trans-ε-viniferin and r2-viniferin from grape cane of the V. vinifera variety Cabernet Moravia, various conditions were studied: different solvents, using powdered versus cut cane material, different extraction times, and one-step or multiple extractions. The largest concentrations found were 6030 ± 680 µg/g dry weight (d.w.) for trans-resveratrol, 2260 ± 90 µg/g d.w. for trans-ε-viniferin, and 510 ± 40 µg/g d.w. for r2-viniferin. The highest amounts of stilbenes (8500 ± 1100 µg/g d.w.) were obtained using accelerated solvent extraction in methanol.

Separation and identification of 1,2,4-trihydroxynaphthalene-1-O-glucoside in Impatiens glandulifera Royle.

Methanolic extract from lyophilized roots of Impatiens glandulifera Royle was analyzed by high performance liquid chromatography using DAD and FLD detection and this revealed one dominant highly fluorescent very unstable substance. The stability of this derivative is strongly dependent on the plant material drying procedure and extraction procedure used. The structure of the substance was established as 1,2,4-trihydroxynaphthalene-1-O-glucoside (THNG) according LC-MS and NMR measurements. When lyophilized plant material was extracted with methanol an almost four times higher amount of THNG was found in the extract, compared to the amount of 2-hydroxy-1,4-naphthoquinone obtained, while in the case of the same lyophilized plant material extracted with water there was no THNG in the extract. The main compounds in this case was 2-hydroxy-1,4-naphthoquinone. In the plant material dried at the laboratory temperature and extracted by methanol there are only traces of THNG.

Degradation of surface-active compounds in a constructed wetland determined using high performance liquid chromatography and extraction spectrophotometry.

Degradation of anionic and nonionic surfactants in a constructed wetland with horizontal subsurface flow was studied using high performance liquid chromatography and extraction spectrophotometry. The ratio of individual homologues of linear alkylbenzene sulfonates (LAS) and the efficiency of their removal were studied. Tridecyl-, dodecyl-, undecyl-, and decylbenzene sulfonates were removed with efficiencies of 92.9%, 84.3%, 64.7%, and 41.1%, respectively. These differences are due to sequential shortening of the alkyl chain in homologues during degradation (the higher homologue can provide the lower one). The formation of sulfophenyl carboxylic acids during ω-oxidation of the alkyl chain followed by successive α- and/or ß-oxidation is also a possible mechanism for removal of LAS. Solid phase extraction using Chromabond® HR-P columns was used for preconcentration of the analytes prior to their determination by HPLC. Methylene blue active compounds were determined using extraction spectrophotometry. The average efficiency of their removal was 84.9% in this case. The efficiency of nonionic surfactant removal (98.2%) was significantly higher in comparison to that for anionic surfactants. The concentration of the endocrine disruptor nonylphenol (a product of nonylphenol polyethoxylate surfactant degradation) determined in the profile of the wetland was beneath the limit of detection (0.4 µg/L). The average outflow concentrations of anionic and nonionic surfactants determined by spectrophotometry were 0.54 and 0.021 mg/L, respectively. The average outflow concentrations of decyl- and tridecylbenzene sulfonates determined by HPLC were 0.195 and 0.015 mg/L. Efficiencies of 86.4% and 92.2% were obtained for removal of organic compounds as indicated by chemical and biochemical oxygen demand (COD(Cr) and BOD(5)). These results demonstrate the suitability of the constructed wetland for degrading surface-active compounds.

Comparison of the Main Constituents in Two Varieties of Proso Millet Using GC-MS.

Proso millet (Panicum miliaceum) is neglected in human nutrition. Thanks to the composition of the grains, millet is suitable for people with celiac disease and it is also useful in the prevention of cardiovascular diseases. For screening the substances in all plant parts of millet via GC-MS, two varieties, Hanacká Mana and Unicum, were used. Substances from the group saccharides, amino acids, fatty acids, carboxylic acids, phytosterols and others were identified in the roots, leaves, stems, and seeds. The highest level of saccharides was found in the stems (83%); amino acids in the roots (6.9%); fatty acids in the seeds (24.6%); carboxylic acids in the roots (3%), phytosterols in the seeds (10.51%); other substances, such as tetramethyl-2-hexadecenol (1.84%) and tocopherols (2.15%), in the leaves; retinal in the roots (1.30%) and squalene in the seeds (1.29%). Saccharides were the dominant group in all plant parts of proso millet followed by fatty acids. The dominant saccharides in all parts of the millet plant were sucrose, fructose and psicose. On the contrary, turanose, trehalose, glucose and cellobiose belonged to the least represented sugars. Additionally, amyrin, miliacin, campesterol, stigmasterol, ß-sitosterol, and others were identified. Varietal variability can be assumed, e.g., in retinal, miliacin or amyrin content.

Ambiguous changes in photosynthetic parameters of Lemna minor L. after short-term exposure to naproxen and paracetamol: Can the risk be ignored?

Non-steroidal anti-inflammatory drugs (NSAID) are recently monitored in the aquatic environment. Naproxen (NPX), paracetamol (PCT) and their transformation products can influence the biochemical and physiological processes at the sub-cellular and cellular levels taking part in the growth and development of plants. This study aimed to compare the effects of NPX and PCT, drugs with different physico-chemical properties, on the growth and photosynthetic processes in Lemna minor during a short-term (7 days) exposure. Although duckweed took up more than five times higher amount of PCT as compared to NPX (275.88 µg/g dry weight to 43.22 µg/g when treated with 10 mg/L), only NPX limited the number of new plants by 9% and 26% under 1 and 10 mg/L, respectively, and increased their dry weight (by 18% under 10 mg/L) and leaf area per plant. A considerable (by 30%) drop in the content of photosynthetic pigments under 10 mg/L treatment by both drugs did not significantly affect the efficiency of the primary processes of photosynthesis. Values of induced chlorophyll fluorescence parameters (F0, FV/FM, ΦII, and NPQ) showed just a mild stimulation by PCT and a negative effect by NPX (by up to 10%), especially on the function of photosystem II and electron transport in both intact duckweed plants and isolated chloroplasts. Lowered efficiency of Hill reaction activity (by more than 10% under 0.1 - 10 mg/L treatments) in isolated chloroplasts suspension proved the only inhibition effect of PCT to primary photosynthetic processes. In intact plants, higher treatments (0.5 - 10 mg/L) by both NPX and PCT induced an increase in RuBisCO content. The results prove that the potential effect of various drugs on plants is hard to generalise.

Sulforaphane Treatment in Children with Autism: A Prospective Randomized Double-Blind Study.

Autism spectrum disorder (ASD) is a pervasive neurodevelopmental disorder with repetitive behaviour which affects interaction and communication. Sulforaphane (SFN), an isothiocyanate abundant in the seeds and sprouts of cruciferous vegetables, has been shown to be effective in alleviating autistic behaviour. We performed a prospective double-blind placebo-controlled study to examine the possible effect of SFN in a paediatric cohort aged three to seven years based on measurements of the Autism Diagnostic Observation Schedule-2 (ADOS-2), the Social Responsiveness Scale-2 (SRS-2), and the Aberrant Behaviour Checklist (ABC). The study consisted of three visits over the duration of 36 weeks (baseline, 18 weeks, and 36 weeks). Twenty-eight of the 40 randomized children completed the study. The mean total raw scores on ABC and SRS-2 improved in both groups, but none of the changes reached statistical significance (ABC: 0 weeks p = 0.2742, 18 weeks p = 0.4352, and 36 weeks 0.576; SRS-2: 0 weeks p = 0.5235, 18 weeks p = 0.9176, and 36 weeks 0.7435). Changes in the assessment of the ADOS-2 subscale scores also did not differ between the two study cohorts (ADOS-2: 0 weeks p = 0.8782, 18 weeks p = 0.4788, and 36 weeks 0.9414). We found no significant clinical improvement in the behavioural outcome measures evaluated in children with ASD aged 3-7 years that were treated with sulforaphane.

Separation and identification of highly fluorescent compounds derived from trans-resveratrol in the leaves of Vitis vinifera infected by Plasmopara viticola.

A method for identification of highly fluorescent compounds in vine leaves infected by Plasmopara viticola was developed using reversed phase liquid chromatography with simultaneous diode array and fluorometric detection. Fluorescent compounds were extracted from leaves with a methanol-water mixture (70:30). Separation by HPLC was performed using a C(18) column and gradient elution with water-acetonitrile mixtures (20-80% of acetonitrile). The main unknown fluorescent compound was identified by line spectral comparison with a standard obtained by UV photoisomerization of trans-resveratrol glucoside, and its structure was confirmed by liquid chromatography-mass spectrometry. Identification and structural elucidation of the fluorescent compound in the leaves of Vitis vinifera allows early detection of Plasmopara viticola invasion.

Lignans Extract from Knotwood of Norway Spruce-A Possible New Weapon against GTDs.

Grapevine trunk diseases (GTDs) pose a major threat to the wine industry worldwide. Currently, efficient biological methods or chemical compounds are not available for the treatment of infected grapevines. In the present study, we used an extract from the knotwood of spruce trees as a biological control against GTDs. Our in vitro trial was focused on the antifungal effects of the extract against the most common GTD pathogens-Cadophora luteo-olivacea, Dactylonectria torresensis, Diaporthe ampelina, Diaporthe bohemiae, Diplodia seriata, Eutypa lata, and Phaeoacremonium minimum. Our in vitro trial revealed a high antifungal effect of the extract against all tested fungi. The inhibition rates varied among the different species from 30% to 100% using 1 mg·mL-1 extract. Subsequently, the efficiency of the extract was supported by an in planta experiment. Commercial grafts of Vitis vinifera were treated with the extract and planted. The total genomic DNA of grapevines was extracted 10 days and 180 days after the treatment. The fungal microbial diversities of the treated/untreated plants were compared using high-throughput amplicon sequencing (HTAS). Treated plants showed 76.9% lower relative abundance of the genus Diaporthe and 70% lower relative abundance of the genus Phaeoacremonium 10 days after treatment. A similar scenario was observed for the genus Cadophora 180 days after treatment, where treated plants showed 76% lower relative abundance of this genus compared with untreated grapevines.

Antifungal and Insecticidal Potential of the Essential Oil from Ocimum sanctum L. against Dangerous Fungal and Insect Species and Its Safety for Non-Target Useful Soil Species Eisenia fetida (Savigny, 1826).

The antifungal and insecticidal effect of the essential oil from Ocimum sanctum L. was evaluated using a model set of harmful organisms hazardous for health and the economy. Toxigenic and plant pathogenic filamentous fungi, including causal agents of human infections, were chosen as exemplary fungal groups-Fusarium verticillioides, Penicillium expansum and Aspergillus flavus. Spodoptera littoralis (African cotton leafworm), Culex quinquefasciatus (Southern house mosquito), the lymphatic filariasis vector and potential Zika virus vector, and the common housefly, Musca domestica were chosen as model insects. Major and minor active substances were detected and quantified using GC/MS analysis. Environmental safety was verified using the non-target useful organism Eisenia fetida. Significant antifungal and insecticidal activity, as well as environmental safety, were confirmed. The essential oil showed the highest efficacy against A. flavus according to MIC50/90, and against S. littoralis larvae according to LD50/90. The monoterpenoid alcohol linalool, t-methyl cinnamate, and estragole as phenylpropanoids were detected as effective major components (85.4%). The essential oil from Ocimum sanctum L. was evaluated as universal and significantly efficient, providing a high potential for use in environmentally safe botanical pesticides.

Factors Influencing Sulforaphane Content in Broccoli Sprouts and Subsequent Sulforaphane Extraction.

Broccoli sprouts contain 10-100 times higher levels of sulforaphane than mature plants, something that has been well known since 1997. Sulforaphane has a whole range of unique biological properties, and it is especially an inducer of phase 2 detoxication enzymes. Therefore, its use has been intensively studied in the field of health and nutrition. The formation of sulforaphane is controlled by the epithiospecifier protein, a myrosinase co-factor, which is temperature-specific. This paper studies the influence of temperature, heating time, the addition of myrosinase in the form of Raphanus sativus sprouts in constant ratio to broccoli sprouts, and other technological steps on the final sulforaphane content in broccoli sprout homogenates. These technological steps are very important for preserving sulforaphane in broccoli sprouts, but there are some limitations concerning the amount of sulforaphane. We focused, therefore, on the extraction process, using suitable ß-cyclodextrin, hexane and ethanol, with the goal of increasing the amount of sulforaphane in the final extract, thus stabilizing it and reducing the required amount sulforaphane needed, e.g., as a dietary supplement.

The Use of a Thermal Process to Produce Black Garlic: Differences in the Physicochemical and Sensory Characteristics Using Seven Varieties of Fresh Garlic.

Black garlic (BG) is a product originating from fresh garlic (FG) and substantially differs in many aspects from FG due to the process called ageing. During this thermal process, the health-promoting properties of FG are enhanced, and the sensory traits are altered. However, very little is known about how the physicochemical properties of different FG varieties affect these properties of BG. Thus, the aim of this study was to investigate the influence of seven FG varieties subjected to the thermal process on the physicochemical parameters of BG. To prepare the BG samples, a fifteen-day ageing process involving a temperature gradient ranging from 30 to 82 °C was used. It was found that the antioxidant activity, the total polyphenol content, and the total soluble solids increased during ageing, while the pH level, moisture content, and lightness decreased in all the garlic varieties. The varieties of garlic differed in the studied traits significantly, both before (FG) and after ageing (BG). In the sensory analysis, significant differences between the BG varieties were observed only in the pleasantness of texture, while the remaining sensory descriptors (pleasantness of color, odor, taste and intensity of the garlic aroma, and overall acceptability) were not affected by variety. The correlations suggest that most of the FG's studied parameters in this study do not correlate with the properties of BG and cannot be used for the prediction of the quality of BG. Additionally, HPLC-MS/MS analysis revealed substantial changes in the composition of low molecular compounds.

Raman microspectroscopy of individual algal cells: sensing unsaturation of storage lipids in vivo.

Algae are becoming a strategic source of fuels, food, feedstocks, and biologically active compounds. This potential has stimulated the development of innovative analytical methods focused on these microorganisms. Algal lipids are among the most promising potential products for fuels as well as for nutrition. The crucial parameter characterizing the algal lipids is the degree of unsaturation of the constituent fatty acids quantified by the iodine value. Here we demonstrate the capacity of the spatially resolved Raman microspectroscopy to determine the effective iodine value in lipid storage bodies of individual living algal cells. The Raman spectra were collected from three selected algal species immobilized in an agarose gel. Prior to immobilization, the algae were cultivated in the stationary phase inducing an overproduction of lipids. We employed the characteristic peaks in the Raman scattering spectra at 1,656 cm(-1) (cis C═C stretching mode) and 1,445 cm(-1) (CH(2) scissoring mode) as the markers defining the ratio of unsaturated-to-saturated carbon-carbon bonds of the fatty acids in the algal lipids. These spectral features were first quantified for pure fatty acids of known iodine value. The resultant calibration curve was then used to calculate the effective iodine value of storage lipids in the living algal cells from their Raman spectra. We demonstrated that the iodine value differs significantly for the three studied algal species. Our spectroscopic estimations of the iodine value were validated using GC-MS measurements and an excellent agreement was found for the Trachydiscus minutus species. A good agreement was also found with the earlier published data on Botryococcus braunii. Thus, we propose that Raman microspectroscopy can become technique of choice in the rapidly expanding field of algal biotechnology.

Thermal stability and bioavailability of bioactive compounds after baking of bread enriched with different onion by-products.

This study investigated the effects of the addition of onion waste fractions into gluten-free (GF) bread to promote its health benefits. 5% of the control (C) GF flour blend was replaced with three waste fractions in the form of: fried onion (FO), dried onion (DO) and onion peel (OP). Antioxidant activity, content of flavonols and total polyphenols of breads increased in the following order: C < FO < DO < OP. No differences were observed in sensory analysis. We found that quercetin glycosides, dimers and trimer in OP-bread, determined according to their mass spectra, decomposed during baking and released free quercetin, which points to their thermal instability. Cross-over study revealed that consumption of OP-bread significantly increased (p < 0.05) antioxidant activity of consumers' blood compared to control bread consumption, indicating good bioavailability of flavonols. Results suggest incorporation of OP into GF bread can increase its biological value with satisfactory sensory acceptance.

Mosquitocidal activities of thyme oils (Thymus vulgaris L.) against Culex quinquefasciatus (Diptera: Culicidae).

Oil samples from four chemotypes of Thymus vulgaris were tested for mosquitocidal activity. Oil compounds were analyzed by gas chromatography-mass spectrometry (GC-MS). The different compositions of the oils varied in their biological activity as well. Larvicidal efficiency was determined for the most significant oil thymol type (T), namely the lowest doses LD(50) 32.9 and 14.2 mg/l for the third and fourth instars, respectively. The oils also showed very good efficiency with respect to mortality and to the percentage of adult emergence upon short-term exposure in water treated by lethal doses of individual oils. Although the larvae were left in treated water for only 5 h, mortality corresponding to LD(50) was determined after 24 h, i.e., between 49% and 63%. Furthermore, mortality increased significantly in relation to time of exposure, and total mortality of the larvae at the end of their development was about 90%. Such significant mortality was also naturally reflected in the total emergence of adults. In the control sample, there was 77% adult emergence from the larvae, whereas in oils, there was only from 5.3% to 16% emergence. The greatest fumigate efficiency was found for T and linalool chemotypes, with an LC(50) for 24 h of exposure being 1.1 and 1.8 mg/l, respectively. Essence T was also most efficient in the tarsal test, with LD(50) of 44 microg/cm(2) and LD(90) of 63 microg/cm(2). High antioviposition efficiency was found in all tested oils.

Contribution to the study of rutin stability in the achenes of Tartary buckwheat (Fagopyrum tataricum).

Tartary buckwheat achenes are rich in rutin. However, its utilisation as a food is low due to the bitter taste of the products because of the transformation of rutin to quercetin. The aim of this study was to determine the stability of rutin and selected flavonoids, which are influenced by the different sample preparations exposing the achenes to higher temperatures (60 °C, 100 °C, and 140 °C), and also to determine the specificity of the rutin degrading enzymes in Tartary buckwheat. The achenes were directly extracted with methanol pre-extracted with hexane, or lyophilised before extraction. The rutin and quercetin contents were determined by HPLC. The rutin-degrading enzymes in Tartary buckwheat achenes were not inactivated after a heat treatment up to 140 °C acting on them for three hours; neither to lyophilisation nor pre-extraction with hexane. The rutin-degrading enzymes probably caused the deglycosylation of kaempferol-3-O-rutinoside, kaempferol-3-O-ß-d-glucoside and piceid. Rutin-degrading enzymes were inactivated with 100% methanol.

Exudation of allelopathic substances in buckwheat (Fagopyrum esculentum Moench).

Root exudates of the common buckwheat, especially phenolic compounds, were studied. Their contents, both in the soil during the growing season and in agar medium during germination, were determined by HPLC and GC-MS. The allelopathic activity of the soil from a buckwheat stand was evaluated, as well. Palmitic acid, squalene, epicatechin, vitexin, a gallic acid derivative, and a quercetin derivative were the main compounds of the agar medium. In the soil, palmitic acid methyl ester, vanillic acid, rutin, a gallic acid derivative, and a 4-hydroxyacetophenone derivative were identified. The effects of vitexin, squalene, epicatechin, 4-hydroxyacetophenone, and vanillic and gallic acids were tested on eight plant species. Inhibitive effects were observed in the cases of 4-hydroxyacetophenone and vanillic and gallic acids. Comparisons of the identified compounds and inhibitive effects of soil extracts indicated that palmitic acid and the gallic acid derivative probably have an important function in the allelopathic root response of buckwheat.