Constituents of the leaves of Peucedanum japonicum Thunb. and their biological activity

In the course of our study on the isolation and structure determination of constituents in tropical plants, we focused on Peucedanum japonicum Thunb., belonging to the family Umbelliferae. In this study, a new C(13) norisoprenoid glucoside, (3S)-O-beta-d-glucopyranosyl-6-[3-oxo-(2S)-butenylidenyl]-1,1,5-trimethylcyclohexan-(5R)-ol (1), and two new phenylpropanoid glucosides, 3-(2-O-beta-d-glucopyranosyl-4-hydroxyphenyl)propanoic acid (3) and methyl 3-(2-O-beta-d-glucopyranosyl-4-hydroxyphenyl)propanoate (4), were isolated from the n-butanol soluble fraction of this plant's leaves, together with five known compounds. The structures of these compounds were determined on the basis of spectroscopic evidence. In addition, all isolated compounds were examined for scavenging activity against 1,1-diphenyl-2-picrylhydrazyl radical and inhibitory activity against mushroom tyrosinase. These results suggested that 2-(4-hydroxy-3-methoxyphenyl)propane-1,3-diol (7) and 3-O-beta-d-glucopyranosyl-2-(4-hydroxy-3-methoxyphenyl)propanol (8) showed an appreciable activity in both assay systems.     

Antioxidant properties of phenolic compounds from Pelargonium reniforme

Flavonoids and hydrolyzable tannins isolated from Pelargonium reniforme were evaluated for their antioxidant ability using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical generating system and a luminol-dependent chemiluminescence assay. In both assays, the polyphenols tested showed higher radical scavenging activities than the reference antioxidant, ascorbic acid (IC50 2.6-32.9 microM vs 40.9 microM in the DPPH test, and 2-25 times stronger effects in the chemiluminescence assay). A comparison of the flavonoids and the tannins showed that the latter have more potential than the former. Structural requirements for marked antioxidant activities of hydrolyzable tannins were the presence of galloyl and hexahydroxydiphenoyl groups, and apparently carbonyl (ester) functionalities in oxidatively modified dehydrohexa-hydroxydiphenoyl moieties. For flavonoids, it appeared that a catechol (3',4'-dihydroxy) element in the B-ring were important determinants and that O-glycosides were more effective than flavone-based C-glucosyls. Conspicuously, introduction of a galloyl group significantly enhanced their potentials. The demonstrated marked antioxidant effects of the polyphenols provide a clue for beneficial effects of P. reniforme in the treatment of liver disorders among several ethnic groups in areas of southern Africa.     

Antioxidant activities of abietane diterpenoids isolated from Torreya nucifera leaves

Investigation on antioxidant compounds from the ethanolic extracts of Torreya nucifera leaves resulted in the isolation of abietane diterpenoids, a known 18-methylesterferruginol (1) and a new 18-dimethoxyferruginol (2). The structures of compounds 1 and 2 were elucidated on the basis of their spectroscopic analyses. Compounds 1 and 2 inhibited the Cu2+-mediated, 2,2'-azobis(2-amidinopropane)hydrochloride-mediated and 3-morpholinosydnonimine-1-mediated low-density lipoprotein (LDL) oxidation in the thiobarbituric acid-reactive substances assay as well as the macrophage-mediated LDL oxidation. Compounds 1 and 2 exhibited the potent antioxidant activities in the conjugated diene production, relative electrophoretic mobility, and apoB-100 fragmentation on copper-mediated LDL oxidation. Compound 1 also suppressed nitric oxide production and inducible nitric oxide synthase expression in lipopolysaccharide-stimulated RAW264.7 cells.     

Antitumor activity of compounds isolated from leaves of Eriobotrya japonica

In a search for possible antitumor agents from natural sources, megastigmane glycosides and polyphenolic constituents isolated from the leaves of Eriobotrya japonica (Rosaceae) were found to inhibit the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced activation of Epstein-Barr virus early antigen in Raji cells. Roseoside and procyanidin B-2 were among the active compounds found in an in vitro assay; these compounds were further assessed for antitumor activity in vivo in a two-stage carcinogenesis assay on mouse skin. Roseoside significantly delayed carcinogenesis induced by peroxynitrite (initiator) and TPA (promoter), and its potency was comparable to that of a green tea polyphenol, (-)-epigallocatechin 3-O-gallate, in the same assay.     

Antioxidant activity of phenolic compounds isolated from Mesona procumbens Hemsl

The antioxidant activity of phenolic compounds isolated from Mesona procumbens Hemsl. (Hsian-tsao) was investigated. Hsian-tsao was extracted with various solvents, and the results showed that the fraction treated with acidic ethyl acetate (pH 2) possessed large amounts of phenolic compounds and a strong antioxidant activity on peroxidation of linoleic acid. The antioxidant activity (inhibition of peroxidation, IP%) of the acidic ethyl acetate of Hsian-tsao extract at 50 microg/mL (98.9%) was stronger than those of 50 microg/mL alpha-tocopherol (78%) and BHA at 10 microg/mL (90%). When fractionated with Amberlite XAD-7 gel chromatography, the acidic ethyl acetate fraction of Hsian-tsao extract was separated into four subfractions (A-D). Subfraction B, with high yield and strong antioxidant activity, was further isolated and purified and then identified as containing protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, caffeic acid, and syringic acid by means of UV, EI-MS, and (1)H and (13)C NMR. The antioxidant capability of isolated compounds was also determined using the thiocyanate system and the erythrocyte ghost system. The results indicate that the phenolic acids could be important antioxidant components in Hsian-tsao, among which caffeic acid with the highest antioxidant activity and the greatest content is most important.     

Antioxidant properties of aroma compounds isolated from soybeans and mung beans

Aroma compounds contained in the extracts of soybean and mung bean that possess antioxidant activity were identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major aroma constituents of soybeans were 1-octen-3-ol (13.699 ppm), maltol (1.662 ppm), phenylethyl alcohol (1.474 ppm), hexanol (1.430 ppm), and gamma-butyrolactone (1.370 ppm). The major aroma constituents of mung beans were hexanol (3.234 ppm), benzyl alcohol (2.060 ppm), gamma-butyrolactone (1.857 ppm), 2-methyl-2-propenal (1. 633 ppm), and pentanol (1.363 ppm). The major aroma chemicals of soybeans and mung beans were examined for antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol showed potent antioxidative activities in two different assays. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited the oxidation of hexanal by 100%, 93%, 84%, and 32%, respectively, for a period of 40 days at the 500 microg/mL level. Eugenol, maltol, benzyl alcohol, and 1-octen-3-ol inhibited malonaldehyde (MA) formation from cod liver oil by 91%, 78%, 78%, and 78%, respectively, at the 160 microg/mL level. The antioxidative activity of eugenol was comparable to that of the natural antioxidant alpha-tocopherol (vitamin E).     

Bioactive aromatic compounds from leaves and stems of Vanilla fragrans.

Alcoholic extracts of leaves and stems of Vanilla fragrans were fractionated with ethyl acetate and aqueous butanol. All three fractions of ethyl acetate, butanol, and water were screened for toxic bioactivity against mosquito larvae. The results of these experiments showed that the fractions from the ethyl acetate and butanol phases were both active in the bioassay. Bioactivity of the ethyl acetate fraction was found to be much greater than that from the butanol fraction in mosquito larvae toxicity. The water phase appeared to contain no substances that impaired mosquito larval growth. Repeated column chromatography of the ethyl acetate fraction on silica gel led to the isolation of 4-ethoxymethylphenol (1), 4-butoxymethylphenol (2), vanillin (3), 4-hydroxy-2-methoxycinnamaldehyde (4), and 3,4-dihydroxyphenylacetic acid (5). Compounds 4 and 5 were isolated from Vanilla species for the first time and 2 has not been reported to have been found in a natural form. 4-Ethoxymethylphenol (1) was the predominant compound, but 4-butoxymethylphenol (2) showed the strongest toxicity to mosquito larvae. The structures of the compounds were determined on the basis of their mass spectra and (1)H or (13)C NMR data.     

Solid-phase microextraction of volatile compounds from the chopped leaves of three species of Eucalyptus

Headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography and ion-trap mass spectrometry has been used to identify biogenic volatile organic compounds present in the headspace of chopped leaves of Eucalyptus (E.) dunnii, E. citriodora, and E. saligna. A simple HS-SPME method entailing 30 min of extraction at 30 degrees C was developed for this purpose. Thirty compounds were identified in the headspace of 60 juvenile chopped Eucalyptus leaves, and another 30 were tentatively identified. The presence of compounds such as (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMNT), (E,E)-alpha-farnesene, (E,E,E)-3,7,11,15-tetramethyl-1,3,6,10,14-hexadecapentaene (TMHP), beta-caryophyllene, alpha-humulene, germacrene D, and beta-cubebene in the headspace of the leaves but not in the essential oils from the same Eucalyptus trees and information about the infochemical roles of some of these compounds in other living plant systems suggest they might play a bioactive role in Eucalyptus leaves.     

Antioxidant activity of flavonoids isolated from young green barley leaves toward biological lipid samples

Natural plant flavonoids, saponarin/lutonarin=4.5/1, isolated from young green barley leaves were examined for their antioxidant activity using cod liver oil, omega-3 fatty acids, phospholipids, and blood plasma. The saponarin/lutonarin (S/L) mixture inhibited malonaldehyde (MA) formation from cod liver oil by 76.47+/-0.11% at a level of 1 micromol and 85.88+/-0.12% at a level of 8 micromol. The S/L mixture inhibited MA formation from the omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) by 45.60+/-1.08 and 69.24+/-0.24%, respectively, at a level of 8 micromol. The S/L mixture inhibited MA formation from the phospholipids lecithin I and II by 43.08+/-0.72 and 69.16+/-2.92%, respectively, at a level of 8 micromol. It also inhibited MA formation from blood plasma by 62.20+/-0.11% at a level of 8 micromol. The antioxidant activities obtained from the S/L mixture were comparable to those obtained from alpha-tocopherol and butylated hydroxy toluene (BHT) in all lipids tested.     

马铃薯茎叶挥发性成分的两种提取方法比较

应用水蒸气蒸馏-溶剂萃取法和微波辅助-溶剂萃取法提取了成熟期马铃薯茎叶挥发性成分。用乙醚为萃取溶剂的水蒸气蒸馏法提取时,以0.533%（m/m）得油率获得精油;以正己烷为溶剂的微波辅助萃取法的最适提取条件为：温度60℃,时间9 min,液料比10︰1（V/m）,在此条件下的得油率为0.528%（m/m）。用GC-MS分析检测出水蒸气蒸馏法获得的精油含有81个成分,解析鉴定出占精油相对含量79.386%的43种物质,醇类化合物（24.789%）为主要成分;微波法获得的精油检测出78个成分,解析鉴定出占总精油82.226%的36种成分,酯类化合物（44.482%）为主要成分。2种提取方法获得的精油成分有很大的差别。     

Effects of enrichment of refined olive oil with phenolic compounds from olive leaves

The possibility of preparing olive oil, with the same nutritional value and stability characteristics found in virgin olive oil, by the enrichment of refined olive oil with olive leaf polyphenols was studied. To obtain antioxidant phenols similar to those found in virgin olive oil, these components were extracted from the leaves of several olive cultivars from the Northern region of Portugal, namely, Carrasca, Ripa, Negruche, Cordovil, Verdeal, Madural, and Bical cultivars, under several conditions. The concentration of a leaf extract required for addition to refined olive oil to obtain the same stability as virgin olive oil was determined. The extract from 1 kg of leaves was sufficient to fortify 50-320 L of refined olive oil to a similar stability as a virgin olive oil sample depending on the metal concentration of the oil, cultivar, and time of the year when the leaves were picked.     

Antioxidant and chemopreventive properties of polyphenolic compounds derived from Greek legume plant extracts

Recently, phytochemical compounds present in legumes have gained a lot of interest because they are considered to be possible chemopreventive agents. In the present study, 14 polyphenolic compounds were extracted and identified from two unique varieties of Leguminosae family plants cultivated in Greece and screened for their antioxidant and chemopreventive properties. Ten polyphenolic fractions, which are mainly mixtures of two compounds and five pure flavonoids, were isolated from the methanolic extracts of aerial plant parts of Vicia faba and Lotus edulis (Leguminosae), respectively. All of these fractions exhibited significant DPPH(*) radical scavenging capacity. Furthermore, they exerted significant protective activity against free radical-induced DNA damage. This activity was more potent against ROO(*) radical-induced DNA damage than against that induced by OH(*) radicals. Finally, they exhibited significant ability to inhibit the activity of the topoisomerase I enzyme. These results imply that the polyphenolic compounds identified in the fractions were responsible of the observed properties of the fractions and the initial extracts and indicate different mechanisms by which these phenolic compounds may act as chemopreventive agents.     

Changes in the concentrations of bioactive compounds in plantain leaves

The plantain is used in herbal medicines and for pasturage. Two cultivars of plantain (Plantago lanceolata L.), Grasslands Lancelot and Ceres Tonic, were sown in spring. Changes in catalpol, aucubin, and acteoside concentrations in the leaves during the growing season and by drying after harvesting were quantitatively determined by high-performance liquid chromatography. The concentration of catalpol was relatively low, fluctuating between 1 and 2% of dry matter during the growing season, and there was no clear-cut seasonal change. From spring to midfall, the aucubin concentration increased from 2.1 to 4.8% in Grasslands Lancelot and from 1.0 to 2.7% in Ceres Tonic. These increases were gradual over the season, except for during midsummer, when aucubin concentrations were relatively constant. The acteoside concentration increased from 3.4 to 7.1% in Grasslands Lancelot and from 1.5 to 4.1% in Ceres Tonic over the course of the growing season, although in the summer it declined steadily to lows of 2.5% in Grasslands Lancelot and 1.9% in Ceres Tonic. Our data suggested that midfall was the appropriate time for harvesting plantain for medicinal use. The concentrations of the bioactive compounds steadily decreased in the initial stages of drying both under natural climatic conditions and at 60 degrees C. The development of processing methods to minimize the loss of bioactive compounds is imperative.     

2,2-Diphenyl-1-picrylhydrazyl radical-scavenging active components from Polygonum multiflorum thunb

An activity-directed fractionation and purification process was used to identify the antioxidative components of Polygonum multiflorum Thunb. (PM). Dried root of PM was extracted with 95% ethanol and then separated into water, ethyl acetate, and hexane fractions. Among these only the ethyl acetate phase showed strong antioxidant activity by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test when compared with water and hexane phases. The ethyl acetate fraction was then subjected to separation and purification using silica gel column chromatography and Sephadex LH-20 chromatography. Three compounds showing strong antioxidant activity were identified by spectral methods ((1)H NMR, (13)C NMR, and MS) and by comparison with authentic samples to be gallic acid, catechin, and 2,3,5, 4'-tetrahydroxystilbene 2-O-beta-D-glucopyranoside.     

Antioxidant activity of the main phenolic compounds isolated from hot pepper fruit (Capsicum annuum L)

Four cultivars (Bronowicka Ostra, Cyklon, Tornado, and Tajfun) of pepper fruit Capsicum annuum L. were studied for phenolics contents and antioxidant activity. Two fractions of phenolics, flavonoids (with phenolic acids) and capsaicinoids, were isolated from the pericarp of pepper fruit at two growth stages (green and red) and were studied for their antioxidant capacity. Both fractions from red fruits had higher activities than those from green fruits. A comparison of the capsaicinoid fraction with the flavonoid and phenolic acid fraction from red fruit with respect to their antioxidant activity gave similar results. Phenolic compounds were separated and quantified by LC and HPLC. Contents of nine compounds were determined in the flavonoid and phenolic acid fraction: trans-p-feruloyl-beta-d-glucopyranoside, trans-p-sinapoyl-beta-d-glucopyranoside, quercetin 3-O-alpha-l-rhamnopyranoside-7-O-beta-d-glucopyranoside, trans-p-ferulyl alcohol-4-O-[6-(2-methyl-3-hydroxypropionyl] glucopyranoside, luteolin 6-C-beta-d-glucopyranoside-8-C-alpha-l-arabinopyranoside, apigenin 6-C-beta-d-glucopyranoside-8-C-alpha-l-arabinopyranoside, lutoeolin 7-O-[2-(beta-d-apiofuranosyl)-beta-d-glucopyranoside], quercetin 3-O-alpha-l-rhamnopyranoside, and luteolin 7-O-[2-(beta-d-apiofuranosyl)-4-(beta-d-glucopyranosyl)-6-malonyl]-beta-d-glucopyranoside. The main compounds of this fraction isolated from red pepper were sinapoyl and feruloyl glycosides, and the main compound from green pepper was quercetin-3-O-l-rhamnoside. Capsaicin and dihydrocapsaicin were the main components of the capsaicinoid fraction. A high correlation was found between the content of these compounds and the antioxidant activity of both fractions. Their antioxidant activities were elucidated by heat-induced oxidation in the beta-carotene-linoleic acid system and the antiradical activity by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) decoloration test. The highest antioxidant activity in the beta-carotene-linoleic acid system was found for trans-p-sinapoyl-beta-d-glucopyranoside, which was lower than the activity of free sinapic acid. Quercetin 3-O-alpha-l-rhamnopyranoside had the highest antiradical activity in the DPPH system, which was comparable to the activity of quercetin. The activities of capsaicin and dihydrocapsaicin were similar to that of trans-p-feruloyl-beta-d-glucopyranoside in the DPPH model system.     

Antioxidant activity of plant extracts containing phenolic compounds.

The antioxidative activity of a total of 92 phenolic extracts from edible and nonedible plant materials (berries, fruits, vegetables, herbs, cereals, tree materials, plant sprouts, and seeds) was examined by autoxidation of methyl linoleate. The content of total phenolics in the extracts was determined spectrometrically according to the Folin-Ciocalteu procedure and calculated as gallic acid equivalents (GAE). Among edible plant materials, remarkable high antioxidant activity and high total phenolic content (GAE > 20 mg/g) were found in berries, especially aronia and crowberry. Apple extracts (two varieties) showed also strong antioxidant activity even though the total phenolic contents were low (GAE < 12.1 mg/g). Among nonedible plant materials, high activities were found in tree materials, especially in willow bark, spruce needles, pine bark and cork, and birch phloem, and in some medicinal plants including heather, bog-rosemary, willow herb, and meadowsweet. In addition, potato peel and beetroot peel extracts showed strong antioxidant effects. To utilize these significant sources of natural antioxidants, further characterization of the phenolic composition is needed.     

Antioxidant properties of ferulic acid and its related compounds

Antioxidant activity of 24 ferulic acid related compounds together with 6 gallic acid related compounds was evaluated using several different physical systems as well as their radical scavenging activity. The radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) decreased in the order caffeic acid > sinapic acid > ferulic acid > ferulic acid esters > p-coumaric acid. In bulk methyl linoleate, test hydroxycinnamic acids and ferulic acid esters showed antioxidant activity in parallel with their radical scavenging activity. In an ethanol-buffer solution of linoleic acid, the activity of test compounds was not always associated with their radical scavenging activity. Ferulic acid was most effective among the tested phenolic acids. Esterification of ferulic acid resulted in increasing activity. The activity of alkyl ferulates was somewhat influenced by the chain length of alcohol moiety. When the inhibitory effects of alkyl ferulates against oxidation of liposome induced by AAPH were tested, hexyl, octyl, and 2-ethyl-1-hexyl ferulates were more active than the other alkyl ferulates. Furthermore, lauryl gallate is most effective among the tested alkyl gallates. These results indicated that not only the radical scavenging activity of antioxidants, but also their affinity with lipid substrates, might be important factors in their activity.     

Countercurrent supercritical fluid extraction and fractionation of high-added-value compounds from a hexane extract of olive leaves

Countercurrent supercritical fluid extraction (CC-SFE) at a pilot scale plant was used for fractionation of high-added-value products from a raw extract of olive leaves in hexane. Compounds found in the raw extract were waxes, hydrocarbons, squalene, beta-carotene, triglycerides, alpha-tocopherol, beta-sitosterol, and alcohols. The CC-SFE extraction process was investigated according to a 2(3) full factorial experimental design using the following variables and ranges: extraction pressure, 75-200 bar; extraction temperature, 35-50 degrees C; and ethanol as modifier, 0-10%. Data were analyzed in terms of extraction yield, enrichment, recovery, and selectivity. Higher extraction yields were attained at 200 bar. For most of the compounds analyzed enrichment was attained at the same conditions, that is, 75 bar, 35 degrees C, and 10% ethanol. Hydrocarbons were usually recovered in the separators, whereas waxes and alpha-tocopherol remain in the raffinate. Selectivity data reveal that alpha-tocopherol is the most easily separable compound. The influence of the experimental factors on the recovery of all the compounds was studied by means of regression models. The best fitted model was attained for beta-sitosterol, with R2 = 99.25%.     

Antioxidant activity and phenolic compounds in selected herbs.

The antioxidant capacities (oxygen radical absorbance capacity, ORAC) and total phenolic contents in extracts of 27 culinary herbs and 12 medicinal herbs were determined. The ORAC values and total phenolic contents for the medicinal herbs ranged from 1.88 to 22.30 micromol of Trolox equivalents (TE)/g of fresh weight and 0.23 to 2.85 mg of gallic acid equivalents (GAE)/g of fresh weight, respectively. Origanum x majoricum, O. vulgare ssp. hirtum, and Poliomintha longiflora have higher ORAC and phenolic contents as compared to other culinary herbs. The ORAC values and total phenolic content for the culinary herbs ranged from 2.35 to 92.18 micromol of TE/g of fresh weight and 0.26 to 17.51 mg of GAE/g of fresh weight, respectively. These also were much higher than values found in the medicinal herbs. The medicinal herbs with the highest ORAC values were Catharanthus roseus, Thymus vulgaris, Hypericum perforatum, and Artemisia annua. A linear relationship existed between ORAC values and total phenolic contents of the medicinal herbs (R = 0.919) and culinary herbs (R = 0.986). High-performance liquid chromatography (HPLC) coupled with diode-array detection was used to identify and quantify the phenolic compounds in selected herbs. Among the identified phenolic compounds, rosmarinic acid was the predominant phenolic compound in Salvia officinalis, Thymus vulgaris, Origanum x majoricum, and P. longiflora, whereas quercetin-3-O-rhamnosyl-(1 --> 2)-rhamnosyl-(1 --> 6)-glucoside and kaempferol-3-O-rhamnosyl-(1 --> 2)-rhamnosyl-(1 --> 6)-glucoside were predominant phenolic compounds in Ginkgo biloba leaves.     

Phytotoxins from the leaves of Ruta graveolens

Bioassay-guided fractionation of the ethyl acetate extract of Ruta graveolens (common rue) leaves led to the isolation of the furanocoumarins 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), and the quinolone alkaloid graveoline as phytotoxic constituents. Graveoline and 8-MOP substantially inhibited growth of Lactuca sativa (lettuce) seedlings and reduced chlorophyll content at 100 microM; this effect was not due to a direct effect on chlorophyll synthesis. Radical growth of L. sativa was inhibited by 10 microM 8-MOP. Graveoline inhibited growth of Lemna paucicostata (duckweed) at 100 microM. This is the first report of the phytotoxic activity of graveoline. Growth of Agrostis stolonifera (bentgrass) was inhibited by 5-MOP at 30 microM. All three compounds substantially reduced cell division in Allium cepa (onion) at or below 100 microM. None of the compounds caused significant cellular leakage of Cucumis sativus (cucumber) cotyledon disks at 100 microM. All three compounds inhibit plant growth, at least partially through inhibition of cell division.