Phytochemical Characterization of Purple Coneflower Roots (Echinacea purpurea (L.) Moench.) and Their Extracts.

Echinacea purpurea is a perennial plant that belongs to the Asteraceae family. It has a wide range of applications mainly in the treatment and prevention of inflammations in the respiratory system. The current study aimed to perform a phytochemical characterization of purple coneflower (Echinacea purpurea) roots and their extracts (water, 40%, 50%, 60% ethanol, and 60% glycerol). Phytochemical characterization was carried out by gravimetric, spectrophotometric, and chromatographic methods. Echinacea roots were characterized by a low lipid (0.8%) content. In contrast, carbohydrates (45%) and proteins (20%) occupied a large part of the dry matter. Amongst the extracts, the highest yield was obtained using water as a solvent (53%). Water extract was rich in protein and carbohydrates as fructans (inulin) were the most abundant carbohydrate constituent. The most exhaustive recovery of the phenolic components was conducted by extraction with 40% ethanol and 60% glycerol. It was found that water is the most suitable extractant for obtaining a polysaccharide-containing complex (PSC) (8.87%). PSC was composed mainly of fructans (inulin) and proteins with different molecular weight distributions. The yield of PSC decreased with an increasing ethanol concentration (40% > 50% > 60%) but the lowest yield was obtained from 60% glycerol extract. The obtained results showed that Echinacea roots contained a large amount of biologically active substances-phenolic components and polysaccharides and that glycerol was equally efficient to ethanol in extracting caffeic acid derivatives from purple coneflower roots. The data can be used for the preparation of extracts having different compositions and thus easily be incorporated into commercial products.

A jojoba (Simmondsia chinensis) seed cake extracts express hepatoprotective activity against paracetamol-induced toxicity in rats.

This study aimed to investigate the hepatoprotective activity of jojoba seed cake extracts against an acute paracetamol (PC) intoxication. Two aqueous extracts from jojoba (Simmondsia chinensis) seed cake, a simmondsin-rich extract (WE), and a simmondsin-hydrolyzed extract (NE) using Viscozyme L enzyme have been prepared and characterized. After enzyme treatment, simmondsin content decreased from 33.0 % to 3.0 % and glucose content increased from 16.2 % to 27.3 % reflecting simmondsin hydrolysis. Both extracts were administered to different rat groups via gavage (0.6 g/kg b.w.) before PC treatment (2 g/kg b.w.) three times a week for 3 weeks. The PC intoxication altered the serum biomarkers, the oxidative status, and the Tumor necrosis factor alpha (TNF-α), Bax and Bcl-2 protein expressions of tested animals. In addition, the histological analysis of liver tissues proved significant injury and hepatocellular necrosis. WE and NE extract showed a relatively high in vitro radical scavenging (ORAC) and averting activities (HORAC) with a polyphenol content of 3.6 % and 2.9 %, respectively. Both extracts showed a powerful in vivo hepatoprotective activity against PC-induced toxicity by improving the hepatocellular antioxidant status and blocking proteins expression (TNF-α, Bax and Bcl-2), involved in inflammation and liver damage. However, the enzymatic treatment improved the hepatoprotective activity of NE despite its lower simmondsin content and lower in vitro antioxidant capacity. This enhancement could be linked to the synergetic effect between the antioxidant components and the new hydrolytic products as glucose, uronic acids, arabinose and simmondsin-aglycons. These results suggest that jojoba waste could be potentially valorized in developing hepatoprotective drugs.

Structural characterization of polysaccharides from Geranium sanguineum L. and their immunomodulatory effects in response to inflammatory agents.

ETHNOPHARMACOLOGICAL RELEVANCE: Geranium sanguineum L. is used for treatment of inflammations, anemia, malignant diseases of the blood-forming organs, diarrhea, respiratory infections, etc. Only flavonoids in root extracts have been elucidated as immunostimulating and anti-inflammatory compounds, and polysaccharides in the herb have not been examined. AIM OF THE STUDY: to compare the chemical features of polysaccharide complexes (PSCs) from leaves (GSL-PSC) and roots (GSR-PSC) of G. sanguineum, as well as their immunomodulatory activities on leukocytes after inflammation, and effects on the growth of different bacteria. MATERIALS AND METHODS: The samples were isolated by water extraction and their structural features were studied by 2D NMR spectroscopy. The stimulatory effects of both PSCs on human leukocytes were analyzed with flow cytometry. Their suppressive activities on the oxidative burst in blood and derived neutrophils against opsonized zymosan and phorbol myristate acetate were investigated. The effects of the samples on viability, NO and interleukin 6 (IL-6) syntheses in RAW264.7 cells after inflammation with lipopolysaccharides (LPS) were tested. The prebiotic and anti-biofilm activities of the PSCs were evaluated. RESULTS: The total carbohydrate content in the samples was significant (73.6-76.8%). GSL-PSC contained pectins, which were rich in homogalacturonan (HG), and smaller amounts of rhamnogalacturonan (RG) type I, decorated by 1,5-α-L-Araf, 1,4- and 1,6-ß-D-Galp chains. GSR-PSC contained starch, followed by pectins with lower HG content and more RG-I regions, substituted by 1 â†’ 3,5-α-L-arabinans and 1 â†’ 3,6-ß-D-galactans. GSL-PSC and GSR-PSC (200 µg/mL) increased monocyte and granulocyte cell counts, but GSR-PSC also elevated T helper and B cell levels in a normal and activated state. GSR-PSC triggered a dose-dependent (50-200 µg/mL) oxidative burst in blood, but alleviated it after inflammation even in blood-derived neutrophils. It was free of LPS, and activated NO and IL-6 productions in RAW264.7 cells better than GSL-PSC, without affecting their viability. Both PSCs (2.0%, w/v) stimulated probiotic co-cultures between Clostridium beijerinckii strains and Lactobacillus sp. ZK9, and inhibited the growth and biofilm formation of Escherichia coli, Streptococcus mutans and Salmonella enterica. CONCLUSIONS: The PSs in G. sanguineum could be involved in the stimulatory effects on blood-forming organs and anti-inflammatory action of aqueous root extracts in case of infections. These PSs should be included in synbiotic foods to support the treatment of inflammations and infections in the gut.

Influence of Gamma Irradiation on Different Phytochemical Constituents of Dried Rose Hip (Rosa canina L.) Fruits.

Gamma irradiation is efficiently applied to many foods, but nevertheless there is a distinct lack of information about the changes of macro- and micronutrients (e.g., carbohydrates, lipids, organic acids, and phenolics) in dried rose hip (RH) fruits. Therefore, in this study, for the first time, the effect of gamma irradiation (10 and 25 kGy) on RH constituents is investigated. Different analytical techniques (GC-FID, HPLC-UV, HPSEC-RID, IR-FT, and SEM) are employed to examine this effect. The irradiation treatment (10 kGy) increased the glucose content by 30% and released cellobiose from RH fruits, thus revealing cellulose destruction. The extractability of total uronic acids increased from 51% (control) to 70.5% (25 kGy-irradiated), resulting in a higher pectin yield (10.8% < 12.8% < 13.4%) and molecular heterogeneity. Moreover, de-esterification was not a major effect of the irradiation-induced degradation of pectin. The sample exposure to the highest dose did not change the content of total carotenoids, ß-carotene, and (un)saturated fatty acids, but it affected the tocopherols levels. Gamma rays had a negligible effect on the phenolic constituents and did not affect ORAC and HORAC antioxidant activity. In conclusion, it can be compromised that the exposition of dried RH is safe and can be successfully applied to decontaminate fruits without affecting their nutritional value and biological activity.

Addition of Medicinal Plants Increases Antioxidant Activity, Color, and Anthocyanin Stability of Black Chokeberry (Aronia melanocarpa) Functional Beverages.

The present study investigates the effect of the addition of medicinal plants, such as lady's mantle, lavender, rosehip, and meadowsweet, on the chemical composition, antioxidant activity, and color intensity of ready-to-drink aronia nectar during pasteurization and long-term storage. Pasteurization caused a significant decrease in anthocyanin content of aronia nectar, which reduced to 20% of the initial value after four months of storage. Herbs provided different protection to aronia anthocyanins that degraded more slowly during the four-month storage compared to pasteurized control without herbs. The addition of medicinal plants enriched aronia nectar with phenolic compounds and increased its antioxidant activity by up to 52% in meadowsweet-aronia nectar. Moreover, it was accompanied by a color intensity magnification due to co-pigmentation of aronia anthocyanins and herbal phenolics. In contrast to anthocyanins, which constantly degraded during the whole period, color intensity began to stabilize after 30 days, demonstrating that co-pigmentation was progressively established during the time and rosehip provided the best stabilization of aronia nectar color. Current research demonstrates for the first time that medicinal plants such as lady's mantle, rosehip, and especially meadowsweet can be used to increase antioxidant activity, color, and anthocyanin stability of black chokeberry functional beverages.

Recent advances on bioactive polysaccharides from mulberry.

Mulberry (Moraceae family), commonly considered as a folk remedy, has a long history of usage in many regions of the world. Polysaccharides regarded as one of the major components in mulberry plants, and they possess antioxidant, antidiabetic, hepatoprotective, prebiotic, immunomodulatory and antitumor properties, among others. In recent decades, mulberry polysaccharides have been widely studied for their multiple health benefits and potential economic value. However, there are few reviews providing updated information on polysaccharides from mulberry. In this review, recent advances in the study of isolation, purification, structural characterization, biological activity and the structure-activity relationship of mulberry polysaccharides are summarized and discussed. Furthermore, a thorough analysis of the current trends and perspectives on mulberry polysaccharides is also proposed. Hopefully, these findings can provide a useful reference value for the development and application of natural polysaccharides in the field of functional food and medicine in the future.

Structural study of a pectic polysaccharide fraction isolated from "mountain tea" (Sideritis scardica Griseb.).

The present study was conducted to investigate the structural characteristics of an acid-extracted polysaccharide fraction from mountain tea. The monosaccharide composition revealed that uronic acids (72.4 mol%) considerably predominated in the fraction, followed by smaller amounts of galactose (14.5 mol%) and glucose (6.2 mol%). The fraction contained mostly a highly methyl-esterified homogalacturonan (HG) - 71 mol%. The pectin had a high molecular weight population (∼60-100 kDa). Enzymatic fingerprinting was employed with a combination of HG degrading enzymes and LC-HILIC-MS, HPAEC, HPSEC to examine the structure in greater detail. Unsaturated oligomers released indicated the presence of large blocks of highly methyl-esterified GalA residues. Furthermore, the presence of blocks of non-esterified GalA residues and partly methyl-esterified and acetylated GalA residues in HG domain was demonstrated. The research findings provide a basis for further investigations regarding biological activity and commercial exploitation of mountain tea.

Phytochemical, nutritional and antioxidant characteristics of whitebeam (Sorbus aria) fruits.

BACKGROUND: Common whitebeam, Sorbus aria (L.) Crantz is an European and Near-Eastern deciduous tree with small red fruits. However, the edible use of this forest fruit is currently not widespread. Also, its nutritional and antioxidative properties remained undiscovered. In this study, the chemical composition and antioxidant properties of common whitebeam fruit were investigated. METHODS: The fruit were collected from Golo Bardo Mountain in Bulgaria. The phytochemical (carotenoids, phenolic compounds, flavonoids), nutritional (moisture, ash, titrable acidity, lipids, proteins, pectin, carbohydrates) and antioxidant activity were evaluated by four assays (DPPH, ABTS, FRAP, and CUPRAC). RESULTS: Sugar analysis demonstrated that only fructose and glucose were detected in the fruit. Pectin content did not exceed 1.30%. Moreover, the fruit had a low lipid content (0.80%). The whitebeam fruit were also characterized as a source of carotenoids (1.69 mg/100 g fresh weight, fw), phenolic compounds (32.42 mg GAE/100 g fw), and flavonoids (20.08 mg QE/100 g fw). Among phenolic acids, only 2,4-dihydrohybenzoic, caffeic, p-coumaric, and sinapic acid were detected. The antioxidant potential of the fruit was from 168.52 mM TE/100 g fw (FRAP) to 244.81 mM TE/100 g fw (CUPRAC assay). CONCLUSIONS: The current research enriched the available information about the nutritional potential and chemical composition of common whitebeam fruit and their low sugar content. The absence of sucrose, together with its high phenolic content, demonstrated the potential of this fruit for future application in food products and supplements.

Structural, rheological and functional properties of galactose-rich pectic polysaccharide fraction from leek.

An acid-extracted polysaccharide from alchohol-insoluble solids of leek was obtained. The sugar composition indicated that galactose and galacturonic acid were the major sugars, followed by small amounts of rhamnose and arabinose. The fraction contained a relatively high methyl-esterified homogalacturonan next to rhamnogalacturonan type I decorated with galactose-rich side chains. The fraction consisted of three high Mw populations, covering the range of 10-100 kDa. Enzymatic fingerprinting was performed with HG/RG-I degrading enzymes to elucidate the structure. The oligomers were analysed using LC-HILIC-MS, HPAEC, and MALDI-TOF MS. The data revealed the presence of GalA sequences, having different patterns of methyl-esterification, RG-I composed of unbranched segments and segments heavily substituted with ß-(1→4)-linked galactan chains of varying length. The rheological study showed the shear-thinning, weak thixotropic, anti-thixotropic, and non-Newtonian behavior of the polysaccharide. The pectin exhibited higher water holding capacity than oil-holding capacity and the fraction did form stable foams at high concentration.

The ancient Thracian endemic plant Haberlea rhodopensis Friv. and related species: A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Haberlea rhodopensis (HR) use dates back to the Thracian and Roman periods. Bulgarians call it Orpheus flower and exploit its leaves for making tea and extracts with detoxifying, tonic, restorative and rejuvenating effects. HR was traditionally applied in wound healing and treatment of cattle diseases. AIM OF THE STUDY: The general aim of the review was to analyze the progress of phytochemical and pharmacological studies on HR, focusing on its radioprotective and immunomodulating effects. MATERIALS AND METHODS: The main source material for the review was collected using several global search engines with the phrase: Haberlea rhodopensis, as well as Bulgarian books and dissertations. RESULTS: HR metabolite profile includes large amounts of free sugars, polyols, polysaccharides (PS), flavonoids, phenolic acids and carotenoids. The radioprotective effect of 70% ethanolic leaf extract (70HREE) is explained by preservation of lymphocytes, other blood cells and testicular tissue from aberration under γ-radiation via stimulation of antioxidant enzymes and neutralization of free radicals. The extract immunomodulating activity results from raised antibody response, stem and neutrophil cell count, complement system activation, anti-tumour and anti-inflammatory effects. The detoxifying, restorative, rejuvenating and wound healing plant properties known to ethnomedicine were supported by radioprotective and immunomodulating studies. CONCLUSIONS: Metabolites of phenolic origin involved in HR resurrection are supposed to contribute to its radioprotective, immunomodulatory, anti-mutagenic and anti-aging effects. However, there is no chemical characterization of 70HREE in the investigations with humans and animals. Structure-activity relationship studies on HR immunomodulating and radioprotective compounds, and on their mode of action are required. They should include not only phenols but PS and other unexplored molecules. The metabolic activity of phagocytes, platelets and lymphocytes triggered by HR extracts has to be examined to elucidate their immunostimulatory potential. HR formulations can be tested in cosmetic, food and medical products as adjuvants to treat infectious, chronic inflammatory and tumour diseases, and especially in patients undergoing radiotherapy.

Co-pigmentation of black chokeberry (Aronia melanocarpa) anthocyanins with phenolic co-pigments and herbal extracts.

The co-pigmentation of black chokeberry (Aronia melanocarpa) anthocyanins with ten phenolic co-pigments was studied. Tested compounds provoked different co-pigmentation effect, manifested by hyperchromic and batochromic shifts. The co-pigmentation was accompanied by a magnification of color intensity and decrease of color hue, both related to a more pleasant color. The hyperchromic effect was the most significant for rosmarinic acid (51.02%), syringic acid (43.24%) and catechin (39.73%). However, it was observed at the highest pigment/co-pigment ratio (1:50), not achievable in plant matter. Targeting the potential practical application of co-pigmentation, we tested eight herbal extracts for their co-pigmentation ability with aronia anthocyanins. The use of herbal extracts led to a significant hyperchromic effect at much lower pigment/co-pigment ratios, compared to pure compounds. The use of selected herbal extracts as co-pigments opens realistic prospects for development of aronia functional foods with improved sensory properties and biological effects, due to enhanced color and anthocyanin stability.

Carotenoids, tocopherols, organic acids, charbohydrate and mineral content in different medicinal plant extracts.

Nettle (Urtica dioica L.), tansy (Tanacetum vulgare L.), bladder campion (Silene vulgaris (Moench) Garcke, waterpepper (Polygonum hydropiper L.), common centaury (Centaurium erythraea Pers.) and rose hip fruit (Rosa canina L. cv. Plovdiv 1) were used for preparation of different water extracts (infusion, decoction and microwave extract) and ethanol (tincture) extracts. Carotenoids (lutein, lycopene and ß-carotene), tocopherols (α-, γ- and δ-), organic acids (ascorbic, malic, fumaric and citric), five macro- and three microelements, sugars and uronic acids content in the obtained extracts were analyzed. Among the investigated plants, stinging nettle, bladder campion and rose hip fruit were evaluated as most potential with respect to bioactive compounds and microelements. The results showed that the selected six medicinal plants and their extracts can be presented as sources of dietary fibers and micronutrients, which may encourage further application as food supplements and beverages as well as to motivate plant use as a dietary alternative in different foods. The present study is a first detailed analysis with respect to sugar content of decoction, infusion and tincture of S. vulgaris.

The common lavender (Lavandula angustifolia Mill.) pectic polysaccharides modulate phagocytic leukocytes and intestinal Peyer's patch cells.

Two pectic (chPS-L1, chPS-L2) and one polyphenolic (chPP-L) fractions were obtained from lavender flowers after boiling water extraction, exhaustive removing of alcohol-soluble molecules and SEC. chPS-L1 (52.4kDa) contains mainly low-acetylated and high-methoxylated homogalacturonans (HG), and smaller rhamnogalacturonan (RG) I backbone fragments rich in 1,3,5-branched arabinan and arabinogalactan (AG) II side chains. chPS-L2 (21.8kDa) contains predominantly similarly esterified HG, followed by RGI with AGII structures and RGII. The prevalence of catechin and epicatechin in chPP-L indicates that they form weak interactions with pectins. chPS-L1 and chPS-L2 enhanced ß2-integrin expression on neutrophils, inducing ROS generation and macrophage NO production. Both the effects on ß2-integrin and high complement fixation activity of chPS-L1 were proposed for its inhibitory action against PMA- and OZP-activated ROS formation. This, together with suppression of NO generation after co-stimulation with chPS-L1 and LPS, suggested anti-inflammatory activity of studied pectins. Lavender polysaccharides expressed intestinal Peyer's patch immunomodulating activity.

Acidic polysaccharide complexes from purslane, silver linden and lavender stimulate Peyer's patch immune cells through innate and adaptive mechanisms.

Three polysaccharide complexes (PSCs) were isolated from the aerial parts of common purslane (Portulaca oleracea L.), and the flowers of common lavender (Lavandula angustifolia Mill.) and silver linden (Tilia tomentosa Moench) by boiling water extraction and ethanol precipitation. The chemical composition and immunomodulating effects of isolated PSCs were characterized. The chemical characterization revealed that the three samples contain mainly pectic polysaccharides. They exhibited ex vivo intestinal immunomodulating activity through the murine Peyer's patch-mediated bone marrow cell proliferation test at 100µg/ml concentration. At the same time, they stimulated ex vivo human blood T-cell populations (CD4+/CD25+ and CD8+/CD25+), phagocytic leukocytes (CD14+ and CD64+ cells) and induced IL-6 production from human white blood cells and Peyer's patch cells. The herbal PSCs stimulated ex vivo ROS production from whole blood phagocytes and showed unspecific in vitro anti-proliferative activity against normal and A549, HeLa and LS180 tumor cells. This is the first report on immunomodulating studies of linden flower pectins and chemical and biological activity characterization of lavender polysaccharides. Our study demonstrates that similarly to purslane, lavender and silver linden herbal materials contain immunomodulating polysaccharides that could be useful for support of compromised immune system.

Isolation and structure elucidation of pectic polysaccharide from rose hip fruits (Rosa canina L.).

A pectic polysaccharide from rose hip (RH) fruits has been obtained by extraction with 1% aqueous citric acid. It was found that the polysaccharide fraction mainly consisted of galacturonic acid (45.5%) next to galactose (5.5%) and arabinose (4.7%). RH pectin is having a relatively high degree of methylesterification (62%) and acetylation (10%) and consists of different molecular weight populations in the range of 10-100kDa. Enzymatic fingerprinting was performed using a combination of pectin lyase (PL) and endo-polygalacturonase. Detailed information about the structure and level of galacturonic acid oligomers released was obtained using LC-HILIC-MS/ELSD and HPAEC. Predominantly, unsaturated and methyl-esterified oligomers (DP 3-5) were released indicating that high proportions of methylesterified 'PL degradable' areas were present within the pectin. The data revealed that homogalacturonan is the main building block of the extracted pectin and consists of long methylesterified/acetylated GalA sequences interspersed with small blocks of non-methyl-esterified GalA units.