Effect of Different Durations of Solid-Phase Fermentation for Fireweed (Chamerion angustifolium (L.) Holub) Leaves on the Content of Polyphenols and Antioxidant Activity In Vitro.

Fireweed has recently been recognized as a plant with high antioxidant potential and phenolic content. Its leaves can be fermented to prepare an infusion with ideal antioxidant activity. The aim of this study was to investigate and to determine the influence of solid-phase fermentation of different durations on the variation of polyphenols in the leaves of fireweed. Laboratory experiments were conducted in 2017-2018. The leaves of fireweed, naturally growing, were fermented for different periods of time: not fermented (control) and fermented for 24 and 48 h. The evaluation of polyphenols and antioxidant activity in leaves was performed using high- performance liquid chromatography (HPLC). Additionally, principal component analysis was used to characterize differences in bioactive compounds between fireweed samples fermented at different durations. Solid-phase fermented leaves were characterized by higher contents of oenothein B, quercetin and benzoic acid but had lower contents of quercetin-3-O-rutinoside, luteolin and chlorogenic and gallic acids. Antioxidant activity in short- (24 h) and long-term (48 h) fermentation (compared to control) gave the highest level of regression in 2017, but in 2018 the effect was observed only with short-term fermentation and control. In conclusion, solid-phase fermentation can be used to modulate biologically active compounds in fireweed leaves.

Aqueous extract from leaves of Ludwigia hyssopifolia (G. Don) Exell as potential bioherbicide.

BACKGROUND: Ludwigia hyssopifolia (G. Don) Exell, one of the problem weeds in some rice-producing countries, was studied to determine its allelopathic potential based on the effects of aqueous extracts of its tissues (leaves, roots and stem) on seedling growth of selected weeds and rice. The major phenolic compound of its leaves was also isolated and characterized. RESULTS: L. hyssopifolia aqueous leaf extract showed significant inhibition of shoot growth and biomass accumulation of weeds (Amaranthus spinosus L., Dactyloctenium aegyptium L., Cyperus iria L.) while maintaining less adverse effects on rice (crop) compared to other aqueous extracts of roots and stem. Phytochemical screening showed that phenols, tannins, flavonoids, terpenoids, saponins and coumarins are found in its leaf aqueous extract. The Folin-Ciocalteu method revealed that its leaves contain 26.66 ± 0.30 mg GAE g-1 leaf. The extract was then acid-hydrolyzed to liberate the phenolics (25 mg phenolics g-1 leaf). The major compound was isolated via preparative thin-layer chromatography using formic acid-ethyl acetate-n-hexane (0.05:4:6) solvent system. It had maximum UV absorption at 272 nm while its Fourier transform infrared spectrum revealed phenol, carboxylic acid and ether functionalities. This also had similar chromatographic mobility when run together with syringic acid in two-dimensional paper chromatography and thin-layer chromatography. CONCLUSIONS: L. hyssopifolia has potential allelopathic activity and its leaf aqueous extract showed the highest phytotoxic activity (P ≤ 0.05) indicating its potential as a bioherbicide. The most probable identity of the major phenolic compound is syringic acid. © 2019 Society of Chemical Industry.

Ellagitannins from the Onagraceae Decrease the Performance of Generalist and Specialist Herbivores.

Phenolics have a role in defenses against herbivores, but the defensive functions of specific groups of phenolics are still poorly understood. For example, ellagitannins (a type of hydrolyzable tannin) are predicted to decrease insect herbivore performance, but the effect of different types of ellagitannins on generalist and specialist herbivores has rarely been assessed. Here, we test the effects of the dominant oligomeric ellagitannins of Oenothera biennis and other Onagraceae on herbivore performance. We fed artificial diets containing between 1 and 100 mg/g of polyphenol fractions comprised of varying amounts and compositions of dimeric oenothein B, the trimeric oenothein A and larger oligomers, to one generalist (Spodoptera exigua) and one specialist (Schinia florida) insect herbivore species. We compared the effects of these ellagitannin fractions on herbivore performance to the effects of artificial diet containing total phenolic extracts from O. biennis, which contained these ellagitannins as well as many additional phenolic metabolites including flavonoid glycosides and caffeic acid derivatives. Both the ellagitannin fractions and O. biennis phenolic extracts had strong negative effects on S. exigua and S. florida performance, with stronger effects on the generalist herbivore. Differences between the effects of the various ellagitannin fractions were small and depended on insect life stage. The defensive effects of these ellagitannins were large, with lethal concentrations as low as 0.1% of the diet. These results highlight the important defensive function of ellagitannins against specialist and generalist herbivores and the need to characterize the effects of these understudied phenolics.

Chemical and Biological Significance of Oenothein B and Related Ellagitannin Oligomers with Macrocyclic Structure.

In 1990, Okuda et al. reported the first isolation and characterization of oenothein B, a unique ellagitannin dimer with a macrocyclic structure, from the Oenothera erythrosepala leaves. Since then, a variety of macrocyclic analogs, including trimeric-heptameric oligomers have been isolated from various medicinal plants belonging to Onagraceae, Lythraceae, and Myrtaceae. Among notable in vitro and in vivo biological activities reported for oenothein B are antioxidant, anti-inflammatory, enzyme inhibitory, antitumor, antimicrobial, and immunomodulatory activities. Oenothein B and related oligomers, and/or plant extracts containing them have thus attracted increasing interest as promising targets for the development of chemopreventive agents of life-related diseases associated with oxygen stress in human health. In order to better understand the significance of this type of ellagitannin in medicinal plants, this review summarizes (1) the structural characteristics of oenothein B and related dimers; (2) the oxidative metabolites of oenothein B up to heptameric oligomers; (3) the distribution of oenotheins and other macrocyclic analogs in the plant kingdom; and (4) the pharmacological activities hitherto documented for oenothein B, including those recently found by our laboratory.

The Role of Leaf Volatiles of Ludwigia octovalvis (Jacq.) Raven in the Attraction of Altica cyanea (Weber) (Coleoptera: Chrysomelidae).

Larvae and adults of Altica cyanea (Weber) (Coleoptera: Chrysomelidae) feed on the rice-field weed Ludwigia octovalvis (Jacq.) Raven (Onagraceae), commonly known as willow primrose, which is considered a biocontrol agent of the weed. Volatile organic compounds from undamaged plants, plants after 4, 12, and 36 h of continuous feeding by A. cyanea larvae or adult females and after mechanical damaging were identified by GC-MS and GC-FID analyses. Twenty nine compounds were identified from undamaged plants. 2Z-Penten-1-ol, geraniol, and 1-tridecanol were present in all plants damaged by larvae. In contrast, feeding by adults caused the release of 2Z-penten-1-ol only after 12 and 36 h; whereas geraniol and 1-tridecanol appeared only after 36 h. Farnesyl acetone was detected after 12 and 36 h of feeding by larvae and after 36 h of feeding by adults. Farnesene was detected after 36 h of feeding by larvae and adults. Linalool was unique after 36 h of feeding by larvae. In Y-shaped glass tube olfactometer bioassays, A. cyanea females were attracted to volatiles after 36 h of feeding by larvae or adults compared to volatiles released by undamaged plants. The insects were attracted to five synthetic compounds: 3-hexanol, α-pinene, linalool oxide, geraniol, and phytol. Synthetic blends were more attractive than individual compounds. Compared to undamaged plants, volatiles released by plants, damaged by conspecific individuals, were more attractive to A. cyanea females, due to elevated emissions of 3-hexanol, α-pinene, linalool oxide, geraniol, and phytol.

Long-chain alkanes and fatty acids from Ludwigia octovalvis weed leaf surface waxes as short-range attractant and ovipositional stimulant to Altica cyanea (Weber) (Coleoptera: Chrysomelidae).

The importance of leaf surface wax compounds from the rice-field weed Ludwigia octovalvis (Jacq.) Raven (Onagraceae) was determined in the flea beetle Altica cyanea (Weber) (Coleoptera: Chrysomelidae). Extraction, thin layer chromatography and GC-MS and GC-FID analyses of surface waxes of young, mature and senescent leaves revealed 20, 19 and 19 n-alkanes between n-C15 and n-C35, respectively; whereas 14, 14 and 12 free fatty acids between C12:0 and C22:0 fatty acids were identified in young, mature and senescent leaves, respectively. Tricosane was predominant n-alkane in young and mature leaves, whilst eicosane predominated in senescent leaves. Heneicosanoic acid, palmitic acid and docosanoic acid were the most abundant free fatty acids in young, mature and senescent leaves, respectively. A. cyanea females showed attraction to 0.25 mature leaf equivalent surface waxes compared with young or senescent leaves in a short glass Y-tube olfactometer bioassay. The insects were attracted to a synthetic blend of 0.90, 1.86, 1.83, 1.95, 0.50 and 0.18 µg ml-1 petroleum ether of hexadecane, octadecane, eicosane, tricosane, palmitic acid and alpha-linolenic acid, respectively, comparable with the proportions as present in 0.25 mature leaf equivalent surface waxes. A. cyanea also laid eggs on a filter paper moistened with 0.25 mature leaf equivalent surface waxes or a synthetic blend of 0.90, 1.86, 1.83, 1.95, 0.50 and 0.18 µg ml-1 petroleum ether of hexadecane, octadecane, eicosane, tricosane, palmitic acid and alpha-linolenic acid, respectively. This finding could provide a basis for monitoring of the potential biocontrol agent in the field.

A Cytotoxic and Anti-inflammatory Campesterol Derivative from Genetically Transformed Hairy Roots of Lopezia racemosa Cav. (Onagraceae).

The genetically transformed hairy root line LRT 7.31 obtained by infecting leaf explants of Lopezia racemosa Cav with the Agrobacterium rhizogenes strain ATCC15834/pTDT, was evaluated to identify the anti-inflammatory and cytotoxic compounds reported previously for the wild plant. After several subcultures of the LRT 7.31 line, the bio-guided fractionation of the dichloromethane-methanol (1:1) extract obtained from dry biomass afforded a fraction that showed important in vivo anti-inflammatory, and in vitro cytotoxic activities. Chemical separation of the active fraction allowed us to identify the triterpenes ursolic (1) and oleanolic (2) acids, and (23R)-2α,3ß,23,28-tetrahydroxy-14,15-dehydrocampesterol (3) as the anti-inflammatory principles of the active fraction. A new molecule 3 was characterized by spectroscopic analysis of its tetraacetate derivative 3a. This compound was not described in previous reports of callus cultures, in vitro germinated seedlings and wild plant extracts of whole L. racemosa plants. The anti-inflammatory and cytotoxic activities displayed by the fraction are associated to the presence of compounds 1-3. The present study reports the obtaining of the transformed hairy roots, the bioguided isolation of the new molecule 3, and its structure characterization.

[Chamerilactone A: one new isocoumarin isolated from Chamerion angustifolium].

A new isocoumarin, chamerilactone A(1) was isolated from the ethanol extract of Chamerion angustifolium with normal phase silica column chromatography, Sephadex LH-20, MCI CHP-20 and semi- preparative HPLC methods. Its structure and stereochemistry were elucidated by spectroscopic methods, including 2D-NMR and optical rotation techniques.

Leptocarposide: a new triterpenoid glycoside from Ludwigia leptocarpa (Onagraceae).

A new triterpenoid bidesmoside (leptocarposide) possessing an acyl group in their glycosidic moiety (1), together with the known luteolin-8-C-glucoside (2) and 1-O-ß-D-glucopyranosyl-(2S,3R,8E)-2-[(2'R)-2-hydroxypalmitoylamino]-8-octadecen-1,3-diol (3) was isolated from the n-butanol-soluble fraction of whole plant of Ludwigia leptocarpa (Nutt) Hara (Onagraceae). Structure of compound 1 has been assigned on the basis of spectroscopic data ((1)H and (13)C NMR, (1)H-(1)H COSY, HSQC, HMBC, and ROESY), mass spectrometry, and by comparison with the literature. This compound was further screened for its potential antioxidant properties by using the radical scavenging assay model 2,2-diphenyl-1-picrylhydrazyl and reveals non-potent antioxidant activities, while compound 2 shows SC50 of 0,038 mM.

[Isolation of pomolic acid from Chamaenerion angustifolium and evaluation of its genotoxicity by microbiological tests].

Fireweed (Chamaenerion angustifolium) lipophilic constituents were investigated by GC-MS analysis. 28 aliphatic and 6 triterpenic acids were found out at first time. Pomolic acid genotoxicity and mutagenicity were analyzed by Ames test and SOS chromotest. Genotoxic and mutagenic properties of pomolic acid were not detected.

Determination of phenolic acids in plant extracts using CZE with on-line transient isotachophoretic preconcentration.

A novel transient ITP-CZE for preconcentration and determination of seven phenolic acids (caffeic acid, cinnamic acid, p-coumaric acid, ferulic acid, protocatechuic acid, syringic acid, and vanilic acid) was developed and validated. Effects of several factors such as control of EOF, pH and buffer concentration, addition of organic solvents and CDs, and conditions for sample injection were investigated. Sample self-stacking was applied by means of induction of transient ITP, which was realized by adding sodium chloride into the sample. The CZE was realized in 200 mM borate buffer ((w)(s)pH 9.2) containing 37.5% methanol, 0.001% hexadimethrine bromide, and 15 mM 2-hydroxypropyl-ß-CD. Under the optimal conditions for analysis, analytes were separated within 20 min. Linearity was tested for each compound in the concentration range of 0.1-10 µg/mL (R = 0.9906-0.9968) and the detection limits (S/N = 3) ranged from 11 ng/mL (protocatechuic acid) to 31 µg/mL (syringic acid). The validated method was applied to the ethanolic extract of Epilobium parviflorum, Onagraceae. The method of SPE was used for the precleaning of the sample.

Antimicrobial, antiparasitic, anti-inflammatory, and cytotoxic activities of Lopezia racemosa.

The present study investigates the potential benefits of the Mexican medicinal plant Lopezia racemosa (Onagraceae). Extracts and fractions from aerial parts of this plant were assessed to determine their antibacterial, antifungal, antiparasitic, anti-inflammatory and cytotoxic activities in vitro. Aerial parts of the plant were extracted with various solvents and fractionated accordingly. Extracts and fractions were tested against a panel of nine bacterial and four fungal species. The antiparasitic activity was tested against Leishmania donovani, whereas the anti-inflammatory activity of the compounds was determined by measuring the secretion of interleukin-6 from human-derived macrophages. The same macrophage cell line was used to investigate the cytotoxicity of the compounds. Various extracts and fractions showed antibacterial, antifungal, antiparasitic, and anti-inflammatory activities. The hexanic fraction HF 11-14b was the most interesting fraction with antimicrobial, and anti-inflammatory activities. The benefit of L. racemosa as a traditional medicinal plant was confirmed as shown by its antibacterial, antifungal and anti-inflammatory activities. To the best of our knowledge, this is the first study reporting the biological activities of L. racemosa, including antiparasitic and anti-inflammatory activities.

Oenothein B suppresses lipopolysaccharide (LPS)-induced inflammation in the mouse brain.

Oenothein B has been recently evaluated for its ability to affect inflammatory responses in peripheral tissues. In this study, we examined its effect on the damage to the central nervous system due to systemic inflammation. For this purpose, ICR mice were injected with an intraperitoneal (i.p.) dose of lipopolysaccharide (LPS; 1 mg/kg mouse). When oenothein B was administered per os (p.o.), it suppressed (1) LPS-induced abnormal behavior in open field; (2) LPS-induced microglial activation in the hippocampus and striatum; and (3) LPS-induced cyclooxygenase (COX)-2 production in the hippocampus and striatum of these mice. These results suggest that oenothein B had the ability to reduce neuroinflammation in the brain during systemic inflammation.

Determination of macrocyclic ellagitannin oenothein B in plant materials by HPLC-DAD-MS: method development and validation.

INTRODUCTION: Oenothein B (OeB) is a dimeric macrocyclic ellagitannin occurring mainly in the genus Oenothera. In the literature no validated HPLC method for quantification of this compound has been reported. OBJECTIVE: To develop and validate an efficient HPLC method for quantification of OeB in plant materials. METHODOLOGY: Extraction conditions of OeB from plant material were optimised. Then the first validated HPLC method for quantification was developed. RESULTS: We have shown that ultrasonic-assisted extraction at 40 °C in 15 min using water are the most optimal conditions for OeB extraction. We have quantified OeB in several plant materials belonging to Oenotheraceae family. The lowest amount of OeB was present in Circaea lutetiana herb (27.64 ± 0.26 mg/g) and the highest was quantified in Epilobium hirsutum aerial parts (72.91 ± 1.38 mg/g). CONCLUSION: The first HPLC method for quantification of OeB in plant materials was developed and validated. We also for the first time optimised the extraction conditions for OeB.

[Fast determination of heavy metal Cu in Ludwigia prostrata leaves using near infrared diffuse spectroscopy].

Heavy metal ions in plants can be determined by using the near-infrared spectral (NIRS) technique, because they combine with the organic molecular groups that have NIRS absorptions. The present article analyzed the fast detection of heavy metal Cu in Ludwigia prostrata leaves by near infrared diffuse spectral technology. Different preprocessing methods were compared, combined with partial least squares (PLS), and the fast detection models of heavy metal Cu in Ludwigia prostrata leaves were established. The results showed that the best model was obtained by PLS with the preprocessing method of average smoothing. The correlation coefficient (r) and root mean square error of calibration(RMSECV) was 0.950 and 5.99 respectively; External validation correlation coefficient (r) and root mean square error of prediction(RMSEP) was 0.923 and 7.38 respectively. The study shows that fast determination of heavy metal Cu in Ludwigia prostrata leaves using near infrared diffuse spectroscopy is feasible.

Chemical composition and in vitro ruminal digestibility of Ludwigia grandiflora.

Ludwigia grandiflora is an amphibious plant that has been designated as a specific alien plant in Japan. Due to the risk of regeneration on land, plants are burned after eradication. Since L. grandiflora is fertile and rich in biomass, this study investigated the use of L. grandiflora as a feed for ruminants. We measured general components, detergent fiber components, and in vitro dry matter digestibility (IVDMD) using rumen microbes for three forms (terrestrial form, emergent form, and floating-leaved form) of L. grandiflora. In addition, concentrations of hazardous metallic elements were also measured from the viewpoint of feed safety. Crude protein, neutral detergent fiber, acid detergent fiber, and acid detergent lignin were contained 10.31-14.62, 27.83-42.28, 20.54-36.05, and 6.94-18.90 (%DM), respectively. IVDMD was 50.75%-68.24%. Toxic elements such as Pb, Cd, and Hg were not detected in all forms. These results suggest that L. grandiflora could be partially used as alternative roughage for ruminants.

The potential and outgoing trends in traditional, phytochemical, economical, and ethnopharmacological importance of family Onagraceae: A comprehensive review.

ETHNOPHARMACOLOGICAL RELEVANCE: The Onagraceae is a widely distributed family of flowering plants comprises about 17 genera and more than 650 species of herbs, shrubs, and trees. Onagraceae also common as willowherb family or evening primrose family is divided into two subfamilies; Ludwigioideae (mainly genus; Ludwigia) and Onagroideae. Family Onagraceae is characterized by its numerous traditional uses as treatment of hormonal imbalances, urinary system ailments, prostate health maintenance, and antimicrobial effects. AIM OF THE STUDY: This review aims to introduce a holistic overview on the phytochemical composition, economical importance and ethnopharmacological value of different species of family Onagraceae. MATERIALS AND METHODS: Literature review was performed using different data bases such as PubMed, Web of Science, Google Scholar and Reaxys searching for articles focused on phytochemical composition, bioactivity and ethnopharmacological history of Onagraceae species. RESULTS: Different species of Onagraceae were reported to have a great variety of phytochemicals including flavonoids, tannins, phenolic acids, triterpenoids, saponins, and volatile/fixed oils. Onagraceae exhibited several health benefits and pharmacological activities including anti-inflammatory, antiarthritic and analgesic, antioxidant, cytotoxic, antidiabetic, and antimicrobial. CONCLUSIONS: Family Onagraceae is an extremely important family with diverse phytochemical composition which enriches their pharmacological importance and hence it's commercial and economical value.

[Haemopoiesis-stimulating and radiomodifying effects of a preparation from a plant Chamaenerion angustifolium].

The experiments as conducted in mice revealed radioprotective therapeutic effect of a preparation made from a plant Chamaenerion angustifolium. The preparation per oral administration in the course of 4 to 8 days post radiation at a dose range of 7.2 to 10.8 Gy promoted an increase in total amounts of blood leukocytes, marrow bone and spleen cells, and also some increase in survival rates among the animals.

Ludwigia octovalvis (Jacq.) raven extract supplementation enhances muscle glycogen content and endurance exercise performance in mice.

Ludwigia octovalvis extract (LOE) is a widely used traditional Chinese herbal medicine. To date, few studies have demonstrated the effect of LOE supplementation on exercise performance, physical fatigue and biochemical profile. The purpose of this study is to evaluate the potential beneficial effects of LOE extract on fatigue and ergogenic functions following physiological challenge. Male ICR mice from 3 groups (n=8 per group) were orally administered LOE for 4 weeks at 0 (vehicle), 61.5 (LOE-1X) or 307.5 (LOE-5X) mg/kg/day. LOE supplementation was able to dose-dependently increase endurance swimming time (P<0.0001) and decrease levels of serum lactate (P=0.0022), ammonia (P<0.0001), creatine kinase (P<0.0001), blood urea nitrogen (P<0.0001) and glucose utilization (P<0.0001) after acute exercise challenge. The glycogen in gastrocnemius muscle also increased with LOE treatment in a dose-dependent manner (P<0.0001). Biochemically, AST, ALT, LDH, CK, BUN, creatinine and UA levels were decreased with LOE treatment. Our study shows that 4-week supplementation with LOE increases muscle glycogen content storage to enhance exercise performance and anti-fatigue effects.

Bioactivities of Flavonoids from Lopezia racemosa.

Lopezia racemosa Cav. (Onagraceae) has been used in Mexican traditional medicine to alleviate stomachache, biliary colic, urine retention, stomach cancer, and skin, dental, buccal, and urinary infections. The objective of this study was to determine the bioactivities of specific parts of the plant to scientifically confirm its traditional use. Aerial parts and flowers were macerated and subsequently extracted with hexane, chloroform, and methanol. This study was focused on the analysis of polar components, and thus the methanolic fractions were selected for further investigations. These fractions were evaluated for their antimicrobial activity using a panel of bacterial Gram-positive and -negative strains, as well as fungal strains, including filamentous fungi and yeasts. In addition, the cytotoxic activity of the extract was assessed by MTT using the human-derived monocytic THP-1 and the normal human fibroblast cell lines. Various fractions showed antimicrobial activity against various pathogens, although the most relevant were against Pseudomonas aeruginosa and Trichophyton mentagrophytes. No inhibition of yeasts was recorded. Only four fractions showed cytotoxic effects when the human-derived THP-1 and fibroblast cells were assessed. The four flavonoids isolated from the extract were luteolin, luteolin-6-C-hexoside, luteolin-8-C-hexoside, and hyperoside. The biological activities presented in this study validate some traditional uses of the plant.