Phytochemical, Pharmacological, and Molecular Docking Study of Dry Extracts of Matricaria discoidea DC. with Analgesic and Soporific Activities.

Pineapple weed (Matricaria discoidea DC.) is a widespread plant in Europe and North America. In ethnomedicine, it is well-known for its anti-inflammatory and spasmolytic activities. The aim of this research was to develop novel methods of M. discoidea processing to obtain essential oil and dry extracts and to investigate their phytochemical compositions. Moreover, the molecular docking of the main substances and the in vivo studies on their soporific and analgesic activities were conducted. The essential oil and two dry extracts from M. discoidea were prepared. A total of 16 phenolic compounds (seven flavonoids, seven hydroxycinnamic acids, and two phenolic acids) in the dry extracts were identified by means of UPLC-MS/MS. In the essential oil, nine main terpenoids were identified by gas chromatography (GC). It was shown that phenolic extraction from the herb was successful when using 70% ethanol in a triple extraction method and at a ratio of 1:14-1:16. The in vivo studies with rodents demonstrated the analgesic activity of the M. discoidea extracts and improvements in the sleep of animals. The dry extracts of M. discoidea did not show any toxicity. The molecular docking analysis showed a high probability of COX-1,2 inhibition and NMDA receptor antagonism by the extracts.

Phytochemical, Technological, and Pharmacological Study on the Galenic Dry Extracts Prepared from German Chamomile (Matricaria chamomilla L.) Flowers.

Galenic preparations of German chamomile are used to treat mild skin diseases, inflammation, and spasms, and they have also been reported to have anxiolytic and sedative effects. The medicinal use of chamomile is well known in ethnomedicine. After obtaining its galenic preparations, there is lots of waste left, so it is expedient to develop waste-free technologies. The aims of this study were to gain knowledge of the ethnomedical status of chamomile in the past and present, develop methods for preparing essential oils and dry extracts from German chamomile flowers using complex processing, reveal the phytochemical composition of such extracts, and verify the analgesic and soporific activity of the extracts. Two methods for the complex processing of German chamomile flowers were developed, which allowed us to obtain the essential oil and dry extracts of the tincture and aqueous extracts as byproducts. A total of 22 phenolic compounds (7 hydroxycinnamic acids, 13 flavonoids, and 2 phenolic acids) were found in the dry extracts by using UPLC-MS/MS. In total, nine main terpenoids were identified in the chamomile oil, which is of the bisabolol chemotype. During the production of chamomile tincture, a raw material-extractant ratio of 1:14-1:16 and triple extraction are recommended for its highest yield. In in vivo studies with mice and rats, the extracts showed analgesic activity and improvements in sleep. The highest sedative and analgesic effects in rodents were found with the dry extract prepared by using a 70% aqueous ethanol solution for extraction at a dose of 50 mg/kg. The developed methods for the complex processing of German chamomile flowers are advisable for implementation into the pharmaceutical industry to reduce the volume of waste during the production of its essential oil and tincture, and to obtain new products.

Phytochemical and Pharmacological Research in Agrimonia eupatoria L. Herb Extract with Anti-Inflammatory and Hepatoprotective Properties.

The most promising plant from the genus Agrimony (Agrimonia L.) of the Rosaceae family for use in medical practice is Agrimonia eupatoria L. Phytochemical and pharmacological research in Agrimonia eupatoria L. herb extract, obtained with using 40% ethanol solution as an extractant were carried out. A total of 11 free and 17 bound monosaccharides, 17 amino acids were found in the studied extract, 9 of which are essential. Gallic and ellagic acids, gallocatechin, epigallocatechin, catechin, epicatechin, and epicatechin gallate were identified in the extract of A. eupatoria by the HPLC method; as well as hydroxycinnamic acids: hydroxyphenylacetate, caffeic, syringic, p-coumaric, ferulic, sinapic, cinnamic and quinic acid; flavonoids: quercetin-3-D-glucoside (isoquercitrin), neohesperidin, naringenin, luteolin were found, and their quantitative content was determined, as well by spectrophotometric methods. The herb extract of A. eupatoria belongs to practically non-toxic substances and has pronounced anti-inflammatory (at a dose of 10.0 mg/kg anti-exudative activity reached a maximum in 5 h (88.17%)) and hepatoprotective activity (at a dose of 25 mg/kg it reduce AlAt level by 1.1 and 1.2 times, respectively; AsAt by 1.2 and 1.1 times, respectively), reduces the level of lipid peroxidation and stabilizes the membrane structures of liver cells. Thus, the herb extract of A. eupatoria is a promising substance for the creation of phytomedicines with anti-inflammatory and hepatoprotective activity.

Quality by Design in optimizing the extraction of (poly)phenolic compounds from Vaccinium myrtillus berries.

Quality by Design was adopted for developing an effective extraction procedure of (poly)phenolic compounds from bilberry (Vaccinium myrtillus L.) fruits, using a pooled sample of berries from different regions of Ukraine. Mechanical solvent extraction, microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) were investigated by screening matrices. Extraction time (Time, from 5 to 15 min), organic solvent type (OS type, methanol, ethanol and acetone), organic solvent percentage (OS%, from 50% to 90%), sample/extractant ratio (S/E ratio, from 0.025 to 0.1 g mL-1), and, only for MAE, extraction temperature (T, from 30 to 60°C), were selected as critical method parameters (CMPs). The spectrophotometric assays total soluble polyphenols (TSP), total monomeric anthocyanins (TMA), and radical scavenging activity (evaluated by the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), the 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid), and the ferric reducing antioxidant power methods) were chosen as critical method attributes (CMAs). The screening procedure allowed for selecting UAE and methanol, while the other CMPs underwent further optimization through Response Surface Methodology. Target values for TSP, TMA and DPPH were selected and the method operable design region (MODR) was defined by means of Monte-Carlo simulations. The optimized conditions, with the corresponding MODR intervals in bracket, were the following: (i) Time, 17 min (15-23 min); OS%, 56% (44-59%); S/E ratio, 0.030 (0.022-0.034) g mL-1. Under these experimental conditions, CMAs values of the pooled sample were the following (n = 3): TSP=4433±176 mg (+)-catechin eq/100 g dry weight (d.w.); TMA=3575±194 mg cyanidin-3-glucoside eq/100 g d.w.; DPPH=273±5 µg DPPH inhib./mg d.w. The optimized extraction method was tested for matrix effect (ME%) in the UHPLC-MS/MS analysis of 15 anthocyanins and 20 non-anthocyanins individual (poly)phenols commonly found in bilberries, as well as for luteolin, sinapic acid, and pelargonidin-3-glucoside, absent in this fruit and therefore added to the extracts as surrogate standards for evaluating apparent recovery (AR%). |ME%| was in any case ≤ 23% and AR% of the surrogate standards in the range 91-95%, confirming the very good performances of the optimized extraction method.

Highbush Blueberry (Vaccinium corymbosum L.) Leaves Extract and Its Modified Arginine Preparation for the Management of Metabolic Syndrome-Chemical Analysis and Bioactivity in Rat Model.

Growing blueberry (Vaccinium corymbosum L., Highbush blueberry) as a berry crop is developing dynamically, especially in warm temperate, subtropical, and tropical regions of the world. When blueberry is cultivated on plantations, the bushes are pruned annually, and tons of leaves become waste. Thus, the aim of the present study was to create a preparation from blueberry leaves, study their chemical composition and determine their potential as a dietary supplement for the prophylactic and correction of the metabolic syndrome. Several schemes for obtaining extracts from blueberry leaves have been developed, including one with addition of arginine. A total of 18 phenolic substances were identified and quantified in the extracts by TLC and HPLC methods. Chlorogenic acid, hyperoside, and rutin were shown to be dominating constituents. Quantitative determination of hydroxycinnamic acid derivatives, flavonoids and other phenolics in the extracts was performed by spectrophotometric method. The extracts administration led to a significant decrease in the level of glucose, insulin and triacylglycerols in blood serum of adult mature inbred rats with insulin resistance induced by the fructose-enriched diet. The most promising one was the extract modified with arginine. The determined hypoglycemic and hypolipidemic activity of chemically standardized extracts from highbush blueberry leaves indicate the potential of this crop residue in utilization as a dietary supplement recommended in prevention of ailments associated with metabolic syndrome.

Characterization of herbal teas containing lime flowers - Tiliae flos by HPTLC method with chemometric analysis.

Linden trees are a source of food products called lime flowers (Tiliae flos), traditionally used in the form of infusion for the treatment of feverish colds and coughs. Lime flowers should include flowers of Tilia cordata Mill, T.x europaea L., and T. platyphyllos Scop. or a mixture of these. The aim of current research was to establish a fast, sensitive HPTLC (high-performance thin-layer chromatography) method that would allow the differentiation of material obtained from five species of lime occurring in Europe. The fingerprints for distinguishing these species were established, as well as a key for identification based on a visual evaluation of chromatograms. The results obtained were also subjected to chemometric analyses. It was shown that each species contains characteristic compounds i.e. linarin that can be used for their identification. The method developed can, in theory, be introduced for the quality control or authentication of linden flowers on the European market.

Immunomodulatory Activity and Phytochemical Profile of Infusions from Cleavers Herb.

Extracts from aerial parts of G. aparine (cleavers) constitute a herbal remedy with monography in British Herbal Pharmacopeia. On the European market, there are several drugs and food supplements consisting of Galium extracts. In folk medicine, cleavers was used topically in Europe, Asia, and the Americas to treat skin diseases. In several remedies, cleavers is also listed as an immunomodulatory active herb influencing the defense response of the human body. The aim of this study was to investigate the immunostimulatory activity and antioxidant potential in vitro of a raw infusion of cleavers and bioactive fractions. The functional activity of lymphocytes in the reaction of the lymphocyte blast transformation (RLBT) method was used for immunomodulatory activity assays and direct scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and hydrogen peroxide (H2O2) was chosen for the examination of antioxidant activity. It was shown that both the raw extract and fractions show significant immunostimulatory and scavenging activities. The obtained data partially justify the traditional use of cleavers as topical remedy for skin infections and for wounds.

Phytochemical Profiles and In Vitro Immunomodulatory Activity of Ethanolic Extracts from Galium aparine L.

Galium aparine L., family Rubiaceae, is a widely spread species in the Galium genus. The herb of G. aparine is part of folk remedies and dietary supplements. In this study, we analyzed the chemical composition and immunomodulatory activities of G. aparine herb ethanolic extracts obtained from the plant material by maceration with 20%, 60% or 96% ethanol. The contents of hydroxycinnamic acid derivatives, flavonoids and polyphenols were determined spectrophotometrically, with extractives and polysaccharides quantified gravimetrically. The qualitative composition was studied using UHPLC-DAD-MS/MS analysis; isolation not previously described in G. aparine quercetin rhamnoglucoside was carried out through column chromatography, and the immunomodulatory activity of extracts was determined in the reaction of lymphocyte blast transformation. Major constitutes of extracts were iridoids, i.e., monotropein, 10-desacetylasperulosidic acid and asperulosidic acid; p-hydroxybenzoic acid; hydroxycinnamic acid derivatives, i.e., 3-O-caffeoylquinic, 5-O-caffeoylquinic, 3,4-O-dicaffeoylquinic, 3,5-O-dicaffeoylquinic, 4,5-O-dicaffeoylquinic acids and caffeic acid derivatives; flavonoids, i.e., rutin, quercetin 3-O-rhamnoglucoside-7-O-glucoside, and isorhamnetin 3-O-glucorhamnoside. Significantly, quercetin 3-O-rhamnoglucoside-7-O-glucoside was first isolated and identified in Galium species so far investigated. All G. aparine herb ethanolic extracts stimulate the transformational activity of immunocompetent blood cells, with 96% ethanolic extract being the most active. The data obtained necessitate further research into the mechanisms of immunomodulatory activity of extracts from G. aparine herb.

The immunomodulatory activity of the extracts and complexes of biologically active compounds of Galium verum L. herb.

The present article discusses the results of an immunomodulatory activity of Galii veri herba, Galium verum herb (Galium verum L., Rubiaceae) fluid water extract and the effect of the polysaccharide and phenolic complexes of the letter on the immunomodulatory activity. The same extract was fractionated into a polysaccharide complex (PSC) and a polyphenolic complex (PPC). In the obtained substances, the contents of hydroxycinnamic derivates, flavonoids and polyphenols were determined spectrophotometrically; polysaccharides were quantified gravimetrically; the immunomodulatory activity of the substances was determined in the reaction of lymphocyte blast transformation. It has been established that the fluid extract of Galium verum herb contains 6.3% polysaccharides, 4.2% hydroxycinnamic derivates expressed as chlorogenic acid, 0.4% flavonoids expressed as rutin, 3.7% polyphenols expressed as gallic acid. PPC contains 4.48% hydroxycinnamic derivates expressed as chlorogenic acid, 0.43% flavonoids expressed as rutin, and 3.95% polyphenols expressed as gallic acid. The lowest immunomodulatory activity was found for PPC. A significantly higher activity was determined for PSC. The highest immunomodulatory activity was established for the fluid extract at a dilution of 1/20, its activity being by 59.4% higher compared with the lymphocyte spontaneous transformation and by 18.5% higher than that of the reference substance PHA. The obtained results enable an assumption of a synergistic effect of PPC and PSC of Galium verum herb fluid extract on the potency of its immunomodulatory activity. Key words: Galium verum L. • immunomodulatory activity • lymphocyte blast transformation • fluid water extract • polysaccharide complex • polyphenols complex.

Evaluation of anti-obesity and lipid-lowering properties of Vaccinium myrtillus leaves powder extract in a hamster model.

Background Vaccinium myrtillus leaves are known to be rich in phenols and have been used in traditional medicine as an antidiabetic remedy. This study evaluated the powder extract of V. myrtillus leaves obtained with the use of L-arginine and myo-inositol for anti-obesity and lipid-lowering potential in hamsters. Methods Standard phytochemical methods were used to determine the total phenolic and total flavonoid contents of the extract. The obesity condition was induced in Syrian hamsters by feeding them with highly palatable fat- and sugar-rich diet (40.3 kcal% fat) for 12 weeks. From the 10th week of diet feeding, the obese hamsters were treated with the powder extract of V. myrtillus leaves (15, 25 and 35 mg/kg/day, respectively) and "Styfimol" (6.2 mg/kg/day of hydroxycitric acid) as a positive control drug. At the end of the treatment period, the biochemical parameters as well as visceral fat mass were determined. Results Vaccinium myrtillus leaves powder extract at 25 and 35 mg/kg/day caused a significant reduction in body weight gain and visceral fat mass in obese hamsters. Serum triacylglycerols, free fatty acids, total cholesterol and low-density lipoprotein cholesterol (LDL-C) levels were also significantly lower. Besides, the hamsters treated with powder extract at 25 and 35 mg/kg/day had the closest intact value ratio of high-density lipoprotein cholesterol and LDL-C compared with positive control animals. Conclusions The results showed that V. myrtillus leaves powder extract is a promising therapeutic agent for the treatment of obesity and obesity-induced diseases.

The immunomodulatory activity of ethanolic extracts from Galium verum L. herb.

The present article discusses the results of the research into the phytochemical profile and immunomodulatory activity of Galii veri herba (Galium verum L., Rubiaceae) ethanolic extracts. The extracts under study were obtained from the plant material by means of maceration technique with 20% aqueous ethanol solution (fluid extract I), 60% aqueous ethanol solution (fluid extract II) and 96% ethanol (fluid extract III) on heating. In the substances obtained, the content of hydroxycinnamic derivatives, flavonoids and polyphenols were determined spectrophotometrically; polysaccharides were quantified gravimetrically; the immunomodulatory activity of the substances under study was determined in the reaction of lymphocyte blast transformation (RLBT). It was established that fluid extract I contained 2.4% polysaccharides, 3.1% hydroxycinnamic derivatives, 0.24% flavonoids, and 2.9% polyphenols; the fluid extract II contained 4.13% hydroxycinnamic derivatives, 0.16% flavonoids, and 3.84% polyphenols; fluid extract III contained 2.7% hydroxycinnamic derivatives, 0.18% flavonoids and 2.7% polyphenols. All the extracts under study possessed a marked stimulant effect on the transformation activity of the immunocompetent blood cells. The highest immunomodulatory activity was established for 96% ethanol extract: the percentage of lymphocytes proliferating in RLBT under the influence of this extract increased by 6.77- 8.04 times in comparison with their spontaneous transformation and by 1.14-1.36 times in comparison with phytohemagglutinin. The results obtained give grounds for further research in the mechanisms of the immunomodulatory activity of the extracts of G. verum herb. Key words: ethanolic extracts Galium verum L. immunomodulatory activity lymphocyte blast transformation.

Research in the chemical composition of the bark of Sorbus aucuparia.

The quantitative contents of 40 carboxylic acids, with the prevalence of hexadecanedioic, lignocerinic, behenic, azelaic, palmitic and benzoic acids, and 39 components of essential oil were determined in the extracts of the bark of Sorbus aucuparia by the GC-MS method. The quantitative contents of 6 phenolic compounds, i.e., chlorogenic, caffeic, and gallic acids, catechin, quercetin, and apigenin-7-glucoside were determined by the HPLC method in the ethanolic extract of the bark of Sorbus aucuparia. Key words: Sorbus aucuparia bark carboxylic acids phenolic compounds essential oil, GC-MS, HPLC.