Changes in Sapwood of Roots of Norway Spruce, Attacked by Fomes annosus

Acetone extracts of sapwood and reaction zone of spruce roots attacked by Fomes annosus, collected in February, June and October, were separated into resinous and phenolic fractions. The fractions were further separated by column, thin layer and gas liquid chromatography, followed by biological tests, using Fomes annosus and other rot fungi. The reaction zone contained quantitatively less light petroleum soluble compounds than the sapwood but more acids. The phenolic content was about ten times higher in the reaction zone than in the sapwood. Nine lignans and one simple phenol (4-methylcatechol) were identified and quantitatively estimated in the reaction zone. The resinous fraction of the extract from the reaction zone as well as some of the lignans and 4-methylcatechol inhibited fungal growth, in some cases followed by detoxification and continued growth. The predominant lignan, hydroxymatairesinol, had no effect on Fomes annosus or five other wood degrading fungi. About 15 unidentified phenols were observed, some of them probably of importance as inhibitors, either alone or in combination with other phenols.     

Red Mexican grapefruit: a novel source for bioactive limonoids and their antioxidant activity

Citrus limonoids have shown to inhibit the growth of cancer in colon, lung, mouth, stomach and breast in animal and cell culture studies. For the first time in the present study, an attempt has been made to isolate antioxidant fractions and five limonoids from red Mexican grapefruit seeds. Defatted seed powder was successively extracted with hexane, ethyl acetate (EtOAc), acetone, methanol (MeOH) and MeOH/water and the extracts were concentrated under vacuum. Radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and total phenolic content were also measured for comparison with the antioxidant capacity in the phosphomolybdenum method for the above extracts. Acetone and MeOH extracts, respectively, showed the highest (85.7%) and lowest (53.3%) radical scavenging activity, at 500 ppm. The total phenolic contents were found to be highest in the acetone extract (15.94%) followed by the MeOH extract (5.92%), ethyl acetate extract (5.54%) and water extract (5.26%). Antioxidant capacity of the extracts as equivalents to ascorbic acid (micromol/g of the extract) was in the order, EtOAc extract > acetone extract > water extract > methanol extract. Furthermore, the EtOAC and acetone extracts were loaded onto silica gel columns to obtain four limonoid aglycons. MeOH fraction was loaded onto a dowex-50 and sepabeads resin column to obtain a limonoid glucoside. The purity of the isolated five compounds was analyzed by HPLC using a C18 column and UV detection at 210 nm. Finally, the structures of the compounds were identified as obacunone, nomilin, limonin, deacetylnomilin (DAN) and limonin-17-beta-D-glucopyranoside (LG) using 1H and 13C NMR studies.     

Knots in trees – A new rich source of lignans

Recent research in our group has revealed that knots, i.e. the branch bases inside tree stems, commonly contain 5–10% (w/w) of lignans. Norway spruce (Picea abies) knots contain as much as 6–24% of lignans, with 7-hydroxymatairesinol (HMR) as the predominant (70–85%) lignan. Some other spruce species also contain HMR as the main lignan, but some spruce species have also other dominating lignans. Most fir (Abies) species contain secoisolariciresinol and lariciresinol as the main lignans. Lignans occur also in knots of pines (Pinus spp.), although in lower amounts than in spruces and firs. Scots pine (Pinus silvestris) knots were found to contain 0.4–3% of lignans with nortrachelogenin as the main lignan. Lignans have been identified also in knots of some hardwoods, although flavonoids are more abundant in hardwoods. Knots are detrimental in the manufacture of pulp and paper and should preferably be removed before pulping. This is possible using a recently developed industrially applicable process called ChipSep. Recent research has also established novel synthetic routes to several lignans, such as matairesinol, secoisolariciresinol, lariciresinol and cyclolariciresinol, starting from hydroxymatairesinol by applying fairly straight-forward chemical transformations. We conclude that wood knots in certain spruce and fir species constitute the richest known source of lignans in nature. The lignans occur in knots in free form and are easily extracted by aqueous ethanol, or even by water. Not only HMR, but also other potentially valuable lignans, could be produced in a scale of hundreds of tons per year by extraction of knots separated from wood chips at pulp and paper mills.     

Characterisation of polyphenols by HPLC-PAD-ESI/MS and antioxidant activity in Equisetum telmateia

The antioxidant activity of an aqueous extract (infusion) and respective ethyl acetate fraction of Equisetum telmateia Ehrh. (Equisetaceae), a plant used in traditional medicine for its anti-inflammatory and diuretic properties, has been evaluated by DPPH, TEAC and TBARS assays. A high and significant antioxidant activity was detected in the ethyl acetate fraction. Analysis of the aqueous extract and the ethyl acetate fraction by HPLC-PAD-ESI/MS allowed the identification of the major phenolic compounds as flavan-3-ol, kaempferol and phenolic acid derivatives. Among the flavan-3-ols, A-type proanthocyanidins and afzelechin derivatives were detected as well as the more common B-type procyanidins, B2 and C1, whose identification was further confirmed by HPLC using detection involving chemical reaction with p-dimethylamino-cinnamaldehyde. The results suggest that the anti-inflammatory activity of E. telmateia could be due, at least in part, to the presence of compounds with antioxidant activity.     

益智的抗氧化作用

本文探讨了益智(Alpinia oxyphylla Miquel)超临界CO2提取物及其渣的水提物、正丙醇提取物和乙酸乙酯提取物的抗氧化作用,测定了总酚含量、黄酮含量、抗氧化力、还原能力、DPPH清除率.结果表明,益智超临界CO2提取物和正丙醇提取物的总酚含量最高,均为5.53%,乙酸乙酯提取物的总酚含量为4.04%,水提物总酚含量最低,为O.89%.抗氧化力与酚含量相关(R2=0.703).四种提取物中黄酮含量顺序为:乙酸乙酯提取物(6.29%)＞丙醇提取物(5.81%)＞水提取物(4.85%)＞超临界CO2提取物(4.70%).在还原能力、清除DPPH自由基和羟自由基方面,乙酸乙酯提取物表现出了很强的抗氧化能力,呈现剂量依赖关系.     

Caesalpinia palmeri: First Report on the Phenolic Compounds Profile,Antioxidant and Cytotoxicity Effect

This study aimed to determine the phenolic compounds profile, antioxidant potential and cytotoxicity of extracts and fractions of Caesalpinia palmeri. Methanolic extracts were generated from C. palmeri berries, stems and flowers. The latter was subjected to liquid-liquid partition, obtaining hexane, ethyl acetate and residues fractions. Results showed that the flower extract and ethyl acetate fraction had a larger concentration of phenolic compounds (148.9 and 307.9 mg GAE/g, respectively), being ellagic acid (6233.57 and 19550.08 μg/g, respectively), quercetin-3-β-glycoside (3023.85 and 8952.55 μg/g, respectively) and gallic acid (2212.98 and 8422.34 μg/g, respectively) the most abundant compounds. Flower extract and ethyl acetate fraction also presented the highest antioxidant capacity on all tested methods (DPPH, ABTS, ORAC and FRAP) and low cytotoxicity against ARPE-19 cells (IC₅₀ >170 μg/mL). C. palmeri possessed high antioxidant potential, associated with the presence of phenolic compounds and low cytotoxicity, suggesting that they could represent an option to counter oxidative stress.     

Effect of Spruce Root Constituents on Extracellular Enzymes of Fomes annosus

Fractions of acetone extracts from heartwood, sapwood and wood from the reaction zone in roots of Norway spruce, attacked by Fomes annosus, were tested concerning their effect on the activity of cellulase, polygalacturonase, aryl-β-glucosidase and laccase, obtained from culture filtrates of the same fungus. A specific inhibition of polygalacturonase was recorded with the resinous fractions. The hydrolysing enzymes were more inhibited by the lignans of the reaction zone in the presence of fungal laccase than in the absence of laccase. The pH dependence of the inhibiting effect of phenols varied among the enzymes. The four enzymes were inhibited by partly different lignan fractions, and synergistic effects between minor compounds were indicated. The phenolic fractions inhibited weakly the biosynthesis of the enzymes. It was obvious that results from enzyme inhibition studies in vitro do not reflect the complicated mutual interrelationships between production and activity of the extracellular fungal enzymes and the chemical constituents of the host in vivo.     

菹草提取物总黄酮、总酚酸和抗氧化活性含量研究

研究菹草提取物的有效成分,测定其总黄酮含量及总酚酸含量,并确定它的乙酸乙酯和石油醚部位的抗氧化活性成分。采用紫外可见分光光度法,以芦丁和没食子酸作为控制材料,测定菹草的石油醚部位及乙酸乙酯部位中总黄酮和总酚酸含量;采用清除DPPH自由基能力测定法、测定总还原能力铁氰化钾法和水杨酸捕捉羟基自由基法来对菹草各组分提取物的抗氧化能力进行研究。菹草中有一定量的黄酮类化合物和酚酸类化合物存在,不同溶剂提取的部位所含有的总黄酮和总酚酸含量是有差异的,石油醚部位的总黄酮含量要高于乙酸乙酯部位的总黄酮含量,并且总酚酸的含量亦是如此;石油醚部位和乙酸乙酯部位的提取物具有清除DPPH、羟基自由基和还原Fe^3+的能力,各部位清除DPPH、羟基自由基的能力和还原Fe^3+的能力随着样品浓度的增大而增大,且乙酸乙酯部位的测定结果均低于石油醚部位。     

Phenolic Compounds of Picea obovata Ledeb. Bark

Russian Journal of Bioorganic Chemistry - Chemical composition of a phenolic complex of the Siberian spruce bark (Picea obovata Ledeb.) and a structure of oligomeric phenolic compounds composing...     

Antioxidant activity of the phenol rich fractions of leaves of Chukrasia tabularis A. Juss

The present study was designed to explore the antioxidant potential of chloroform, ethyl acetate, n-butanol and water fractions of 80% methanol extract of leaves Chukrasia tabularis by 2,2'-diphenyl-1-picrylhydrazyl (DPPH), deoxyribose degradation (non-site specific and site specific), reducing power and DNA nicking assays. The different fractions showed significant activities in all the free radical scavenging tests and these findings have also shown direct relationship between antioxidant activity and phenolic content. Among the fractions, ethyl acetate fraction exhibited highest inhibition of 93.14%, 89.99%, 87.04% in DPPH, non-site specific and site specific deoxyribose degradation assays, respectively and 91.20% reduction of ferricyanide to give Prussian blue coloured complex in reducing power assay at maximum concentration tested. This preliminary study indicates the antioxidant activity of the leaves of Chukrasia tabularis, and moreover the results showed correlation with the amount of phenolic content present in different fractions.     

Antioxidant properties of methanol extract and its solvent fractions obtained from selected Indian red seaweeds

In vitro antioxidant activities of three selected Indian red seaweeds - viz., Euchema kappaphycus, Gracilaria edulis and Acanthophora spicifera were evaluated. Total phenolic content and reducing power of crude methanol extract were determined. The antioxidant activities of total methanol extract and five different solvent fractions (viz., petroleum ether (PE), ethyl acetate (EA), dichloromethane (DCM), butanol (BuOH) and aqueous) were also evaluated. EA fraction of A. spicifera exhibited higher total antioxidant activity (32.01 mg ascorbic acid equivalent/g extract) among all the fractions. Higher phenolic content (16.26 mg gallic acid equivalent/g extract) was noticed in PE fraction of G. edulis. Reducing power of crude methanol extract increased with increasing concentration of the extract. Reducing power and hydroxyl radical scavenging activity of E. kappaphycus were higher compared to standard antioxidant (alpha-tocopherol). The total phenol content of all the seaweeds was significantly different (P<0.05). In vitro antioxidant activities of methanol extracts of all the three seaweeds exhibited dose dependency; and increased with increasing concentration of the extract.     

Antioxidant and anti-inflammatory activity of extract obtained from Aspergillus candidus MTCC 2202 broth filtrate

Antioxidant potential of Aspergillus candidus MTCC 2202 broth filtrate extract was studied using different antioxidant models, whereas anti-inflammatory potential was studied using carrageenan-induced rat paw oedema model. The ethyl acetate extract at 1000 microg/ml showed maximum scavenging activity of the stable radical 1,1-diphenyl,2-picryl hydrazyl upto 96.65% (IC50=430.36 microg/ml) and scavenging of the radical cation, 2,2-azinobis-(3-ethylbenzothiazoline-6-sulphonate) upto 92.25% (IC50=606.29 microg/ml) at the same concentration. The extract had good reducing power, however showed moderate inhibition for conjugated dienes and thiobarbituric acid reactive acid substances (59.56 and 51.45%). The total phenolic content of various extracts of A. candidus broth filtrate was measured and a correlation between radical scavenging activities of extracts with total phenolic content was observed. The ethyl acetate extract (125 mg/kg ip) showed significant anti-inflammatory activity in carrageenan-induced rat paw oedema model. The exhibited antioxidant activity of ethyl acetate extract of A. candidus broth filtrate was comparable with BHA and ascorbic acid, while anti-inflammatory activity was comparable with standard diclofenac sodium.     

Phenolic profile,biological activities and fraction analysis of the medicinal halophyte Retama raetam

Retama raetam is a medicinal and aromatic plant present in the humid to the arid bioclimatic regions of Tunisia. In this work, we investigated R. raetam shoots antioxidant and antimicrobial activities and its natural antioxidant contents obtained from four fractions (petroleum ether, acetone 60%, ethyl acetate and water). Results showed that the ethyl acetate fraction exhibits the highest antioxidant activity as compared to the other ones. In fact, IC₅₀ values of ethyl acetate extract were equal to 33.5, 500 and 1380 μg/ml (DPPH and ABTS radicals scavenging activity and reducing power, respectively). Accordingly, this fraction presented the highest total polyphenol and flavonoid contents (401 mg GAE/g DR and 33.21 mg CE/g DR, respectively). Moreover, RP-HPLC analysis showed that syringic acid and coumarin were the major phenolic compounds. Furthermore, this moderately polar fraction showed considerable antibacterial properties against human pathogen strains especially against Escherichia coli and Bacillus cereus. Finally, fractionation allows the identification of R. raetam most active molecules and therefore the optimization of their utilization. Our findings pointed out the appropriate solvent for extracting R. raetam potent phenolics which might provide a rich and novel source of natural antioxidants as food additives replacing synthetic ones in food industry.     

Antioxidant activities and phytochemical characteristics of extracts from Acacia confusa bark

The present study was carried out to evaluate the antioxidant activities of bark extract of Acacia confusa and some of the isolated constituents from its ethyl acetate (EtOAc) fraction in various in vitro systems to gain mechanistic insights. Results from antioxidant assays together with authentic antioxidant standards revealed that EtOAc fraction showed strong superoxide radical scavenging activity, reducing power, and ferrous ion-chelating ability. Following an in vitro antioxidant activity-guided fractionation procedure, 16 constituents including 12 benzoic acids, three cinnamic acids and one lignans were isolated and identified from the EtOAc fraction. We also evaluated the structure-activity relationships of benzoic and cinnamic acid derivatives. Results obtained indicated that the bark extracts and the derived phytochemicals from A. confusa have a great potential to prevent disease caused by the overproduction of radicals and also it might be used as a potential source of natural antioxidant agent.     

Isolation and Identification of a Novel Antioxidant with Antitumour Activity from <Emphasis Type="Italic">Serratia ureilytica</Emphasis> Using Squid Pen as Fermentation Substrate

The antioxidant activity of the culture supernatant of Serratia ureilytica TKU013 with squid pen as the sole carbon/nitrogen source was assessed by three methods, and the phenolic contents were assayed. The supernatant with the highest antioxidant activity was further purified by liquid–liquid partition, revealing the ethyl acetate extract exhibited the strongest antioxidant activity and the highest total phenolic content. Eight fractions were retrieved from silica gel column chromatography of this extract, designated F1–F8. F4 was found to possess the strong antioxidative activity and the highest total phenolic content and also exhibited strong cytotoxic activities against two different tumoural cell lines. A new compound (Serranticin) with antioxidant and antitumor activity was obtained from F4. The structure of Serranticin is analogous to that of siderophores (hexacoordinated catecholamine), which are iron chelators. As such, Serranticin has the potential for use as a deferration agent in various iron overload diseases.     

Enzyme inhibition and radical scavenging activities of aerial parts of Paeonia emodi Wall. (Paeoniaceae)

The ethanolic extract derived from aerial parts of an indigenous medicinal plant Paeonia emodi was screened for enzyme inhibition activities against Urease (jack bean and Bacillus pasteurii) and α-Chymotrypsin. The extract was also investigated for its radical scavenging activity using DPPH assay. The crude extract was found to possess significant enzyme inhibition activities against jack bean (74%) and Bacillus pasteurii (80%) urease and a moderate activity (54%) against α-Chymotrypsin. The extract also displayed excellent (83%) radical scavenging activity. On the basis of these results, the crude extract was subsequently fractionated into n-hexane, chloroform, ethyl acetate, n-butanol and water fractions and tested independently for the aforesaid activities. Significant inhibitory activity against urease enzyme was observed for the ethyl acetate, n-butanol and water fractions while the n-hexane and chloroform fractions were devoid of any such activity. In the α-Chymotrypsin enzyme inhibition studies the activity was concentrated into the ethyl acetate fraction. All the fractions displayed potent radical scavenging activity. The crude extract and fractions thereof were also subjected to total phenolic content determination. A correlation between radical scavenging capacities of extracts and total phenolic content was observed in the majority of cases.     

Variation of stilbene glucosides in bark extracts obtained from roots and stumps of Norway spruce (Picea abies [L.] Karst.)

Acetone-soluble compounds found in different root zones and stumps of Norway spruce (Picea abies [L.] Karst), which were grown on either peatland or a mineral soil site, were studied. Samples from stumps and roots of different sizes and ages were collected a day after the trees were felled. The wood and bark of stumps and three zones of the roots were separated and extracted with acetone in an ultrasonic bath. Extracts were silylated and analysed by gas chromatography–mass spectrometry. The stilbene glucosides astringin and isorhapontin were major compounds in the spruce bark samples. The resveratrol glucoside piceid and the flavonoid catechin were also extracted from spruce bark. We also found the lignan hydroxymatairesinol in some wood extracts. Total concentrations of stilbene glucosides in bark of stumps and different root zones varied between 0.53 and 8.29 % (w/w, dry weight) with isorhapontin being the major compound. Isorhapontin concentrations were highest in the spruce samples grown on mineral soil. The bark of the roots close to the stem is a rich source of stilbenes for commercial utilisation.     

吸附层析法制备低聚原花青素

以AB 8大孔吸附树脂对葡萄籽原花青素粗提物进行了分级分离 ,所得乙酸乙酯洗脱产物总酚含量为 91% ,经高效液相色谱检测 ,酚类物质中单体约 2 8% ,低聚原花青素占 6 4 % ,多聚体仅为 8%。     

Enzyme inhibition and radical scavenging activities of aerial parts of Paeonia emodi Wall (Paeoniaceae)

The ethanolic extract derived from aerial parts of an indigenous medicinal plant Paeonia emodi was screened for enzyme inhibition activities against Urease (jack bean and Bacillus pasteurii) and alpha-Chymotrypsin. The extract was also investigated for its radical scavenging activity using DPPH assay. The crude extract was found to possess significant enzyme inhibition activities against jack bean (74%) and Bacillus pasteurii (80%) urease and a moderate activity (54%) against alpha-Chymotrypsin. The extract also displayed excellent (83%) radical scavenging activity. On the basis of these results, the crude extract was subsequently fractionated into n-hexane, chloroform, ethyl acetate, n-butanol and water fractions and tested independently for the aforesaid activities. Significant inhibitory activity against urease enzyme was observed for the ethyl acetate, n-butanol and water fractions while the n-hexane and chloroform fractions were devoid of any such activity. In the alpha-Chymotrypsin enzyme inhibition studies the activity was concentrated into the ethyl acetate fraction. All the fractions displayed potent radical scavenging activity. The crude extract and fractions thereof were also subjected to total phenolic content determination. A correlation between radical scavenging capacities of extracts and total phenolic content was observed in the majority of cases.     

Flavonoids, triterpenoids and a lignan from Vitex altissima

A new tetrahydrofuranoid lignan, altissinone (1) and a new acylated flavone C-glucoside, 2'-O-p-hydroxybenzoylorientin (2), were isolated in addition to several known triterpene acids and flavonoids from the ethyl acetate extractives of the leaves of Vitex altissima. The structures of the compounds were established based on interpretation of high resolution NMR (HMQC, HMBC and NOESY) spectral data. The ethyl acetate extract exhibited significant anti-inflammatory activity in rat paw edema model. The flavonoids and triterpene acids showed moderate antioxidant and 5-lipoxygenase enzyme inhibitory activities, respectively.