Identification of new dicaffeoylquinic acids from Chrysanthemum morifolium and their antioxidant activities

Two new dicaffeoylquinic acids, 3,5-dicaffeoyl- epi-quinic acid (1) and 1,3-dicaffeoyl- epi-quinic acid (2), were isolated from Chrysanthemum morifolium Ramar together with six known dicaffeoylquinic acid derivatives and three flavonoids. The structures of the new compounds were elucidated using of spectroscopic methods. These compounds were assessed for antioxidant activities in the DPPH radical and superoxide anion radical scavenging systems. Most of the isolates showed strong antioxidant activities in these assay systems. Two new compounds showed potent superoxide anion radical scavenging activity (IC50 = 2.9 +/- 0.1 for 1 and 2.6 +/- 0.4 microg/mL for 2, respectively) in the xanthine/xanthine oxidase system as compared to quercetin and also showed potent DPPH radical scavenging activity (IC50 = 5.6 +/- 0.1 for 1 and 5.8 +/- 0.2 microg/mL for 2, respectively).     

The isolation and antioxidative effects of vitexin fromAcer palmatum

Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental factors are critical players in cellular damage and aging. In order to develop a new anti-photoaging agent, this work focused on the antioxidant effects of the extract of tinged autumnal leaves of Acer palmatum. One compound was isolated from an ethyl acetate soluble fraction of the A. palmatum extract using silica gel column chromatography. The chemical structure was identified as apigenin-8-C-beta-D-glucopyranoside, more commonly known as vitexin, by spectral analysis including LC-MS, FT-IR, UV, 1H-, and 13C-NMR. The biological activities of vitexin were investigated for the potential application of its anti-aging effects in the cosmetic field. Vitexin inhibited superoxide radicals by about 70% at a concentration of 100 microg/mL and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals by about 60% at a concentration of 100 microg/mL. Intracellular ROS scavenging activity was indicated by increases in dichlorofluorescein (DCF) fluorescence upon exposure to UVB 20 mJ/cm2 in cultured human dermal fibroblasts (HDFs) after the treatment of vitexin. The results show that oxidation of 5-(6-)chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) is inhibited by vitexin effectively and that vitexin has a potent free radical scavenging activity in UVB-irradiated HDFs. In ROS imaging using a confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our findings suggest that vitexin can be effectively used for the prevention of UV-induced adverse skin reactions such as free radical production and skin cell damage.     

乌梅抗氧化作用的实验考察

目的:研究乌梅提取物抗氧化作用。方法:通过检测还原力和清除DPPH、ABTS和超氧阴离子自由基的能力,分析和评价乌梅提取物的抗氧化活性。结果:乌梅醇提物对DPPH、ABTS自由基的IC50分别为0.500 mg.mL-1及0.253 mg.mL-1;乌梅水提物对DPPH、ABTS自由基的IC50分别0.510 mg.mL-1及0.500 mg.mL-1;乌梅醇提物还原能力大于乌梅水提物,且乌梅醇提物对超氧阴离子自由基的清除作用显著,其清除作用优于乌梅水提物及对照品BHT。结论:乌梅提取物有不同程度的抗氧化活性,其中以醇提物抗氧化作用最强。     

茉莉花挥发油清除自由基活性的研究

目的：从清除1,1－二苯基－2－苦苯肼自由基（DPPH?）、羟自由基（? OH）、超氧阴离子自由基（O2?－）3个方面,研究茉莉花挥发油清除自由基活性。方法采用水蒸气蒸馏法提取茉莉花挥发油,从其清除DPPH?、? OH和O2?－能力3个方面进行考察,并以常用抗氧化剂L－抗坏血酸为对照作比较,通过计算半数抑制浓度（ IC50）评价茉莉花挥发油清除,在供试质量浓度范围内,呈一定量效关系,其 IC50分别为153．548μg?mL －1、56．481μg? mL －1和133．586μg? mL －1。结论茉莉花挥发油具有良好的清除自由基活性,是一种新型天然抗氧化物质来源。     

滇黄芩茎叶乙醇提取物及其不同溶剂萃取部位的抗氧化和降脂活性研究

目的:研究滇黄芩茎叶乙醇提取物及其不同溶剂萃取部位的抗氧化活性和降脂活性。方法:取滇黄芩茎叶用95%乙醇回流提取,得乙醇提取物;取上述提取物,依次用石油醚、乙酸乙酯、正丁醇萃取,回收溶剂得到不同溶剂萃取部位。以维生素C(Vc)为阳性对照,采用羟基自由基、超氧阴离子自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法检测滇黄芩茎叶乙醇提取物、石油醚萃取物、乙酸乙酯萃取物、正丁醇萃取物的体外抗氧化活性并计算半数抑制浓度(IC50)。以脂肪乳建立脂肪变性L02肝细胞模型,以非诺贝特(20μg/m L)为阳性对照,考察高、低质量浓度(100、50μg/mL)滇黄芩茎叶乙醇提取物、乙酸乙酯萃取物、正丁醇萃取物对细胞中TC、TG含量的影响。结果:对羟基自由基的清除能力强弱排序为滇黄芩茎叶正丁醇萃取物>乙酸乙酯萃取物>Vc>乙醇提取物>石油醚萃取物,IC50分别为0.15、0.17、0.35、0.75、1.17 mg/mL;对超氧阴离子自由基的清除能力强弱排序为Vc>正丁醇萃取物>乙酸乙酯萃取物>乙醇提取物>石油醚萃取物,IC50分别为0.034、0.55、0.75、3.32、3.73 mg/mL;对DPPH自由基的清除能力强弱排序为Vc>正丁醇萃取物>乙酸乙酯萃取物>乙醇提取物>石油醚萃取物,IC50分别为0.003 2、0.028、0.033、0.048、0.057 mg/mL。滇黄芩茎叶乙醇提取物、乙酸乙酯萃取物和正丁醇萃取物对脂肪变性L02肝细胞中TC、TG含量均有显著的降低作用(P<0.01),其降脂能力强弱排序为正丁醇萃取物(低浓度)≈非诺贝特>乙酸乙酯萃取物(高浓度)>乙醇提取物(高浓度)>正丁醇萃取物(高浓度)>乙酸乙酯萃取物(低浓度)>乙醇提取物(低浓度)。结论:滇黄芩茎叶乙醇提取物、石油醚萃取物、乙酸乙酯萃取物、正丁醇萃取物均有一定的抗氧化活性和降脂活性(除石油醚萃取物外),其中正丁醇和乙酸乙酯萃取物的抗氧化活性和降脂活性较高。     

Evaluation of the antioxidant activity of soybean extract by different in vitro methods and investigation of this activity after its incorporation in topical formulations.

Chemoprevention by natural products is an emerging therapeutic approach for free radical-mediated diseases including cancer. This is a consequence of its wide applicability and acceptance. In the present study, the antioxidant activity of the soybean extract (Isoflavin Beta) and of formulations added with this extract were evaluated using stable free radical 2,2-diphenyl-1-pycrylhydrazyl (DPPH*) and deoxyribose as well as the lipid peroxidation inhibition assays. For all the assays the extract showed a dose-dependent activity, and IC50 of 21.03 microg/mL in lipid peroxidation inhibition, 161.8 microg/mL in DPPH*, and 33.5 ng/mL in hydroxyl radical scavenging assay. The antioxidant activity of the extract added in the formulations could not be assessed using the deoxyribose assay. However, the lipid peroxidation inhibition and DPPH* scavenging assays could be successfully applied for the antioxidant activity evaluation of the formulations added with soybean extract to protect the skin against free radicals, which can be generated by the ultraviolet radiation exposure.     

芙蓉菊多糖体外抗氧化活性研究

目的研究芙蓉菊多糖的体外抗氧化活性。方法采用1,1-二苯基苦基苯肼(DPPH)自由基、还原力、超氧阴离子、羟自由基4种体外抗氧化模型研究芙蓉菊多糖的抗氧化活性,用维生素C作对照。结果芙蓉菊粗多糖对DPPH自由基、还原力、超氧阴离子、羟自由基均有明显的清除能力,其中DPPH、超氧阴离子、羟自由基的EC50值分别为0.273、0.669、0.594mg/mL,对羟自由基清除能力强于维生素C。芙蓉菊多糖对自由基清除率与其质量浓度存在着明显的量效关系。结论芙蓉菊多糖具有良好的体外抗氧化活性,作为天然抗氧化剂和自由基清除剂有进一步研究的价值。     

红景天中8种成分体外抗氧化作用的比较

目的比较性研究红景天中8种苷及醇类化合物———芦丁苷、红景天苷、异槲皮苷、酪醇、络塞琳、熊果苷、络塞维、肉桂醇红景天的体外抗氧化作用。方法测定8种成分对DPPH自由基、超氧阴离子自由基、一氧化氮自由基、羟自由基的清除能力,以及总抗氧化能力和抑制脂质过氧化反应的能力。结果除清除羟自由基实验外,异槲皮苷和芦丁苷作用明显强于其他化合物,其中在清除DPPH、超氧阴离子能力和抑制脂质过氧化实验中的IC50分别为5.72和10.46,23.63和36.39,10.78和56.49 mg.L-1;而在清除羟自由基实验中,酪醇和红景天苷的作用则强于其他化合物,IC50分别为10.84和18.97 mg.L-1。结论红景天中苷及醇类成分对多种自由基具有清除作用,有良好的综合抗氧化性和开发应用前景。     

Antioxidant properties of Aller-7, a novel polyherbal formulation for allergic rhinitis

Allergic rhinitis, a frequently occurring immunological disorder affecting men, women and children worldwide, is a state of hypersensitivity that occurs when the body overreacts to a substance such as pollen, mold, mites or dust. Allergic rhinitis exerts inflammatory response and irritation of the nasal mucosal membranes leading to sneezing; stuffy/runny nose; nasal congestion; and itchy, watery and swollen eyes. A novel, safe polyherbal formulation (Aller-7/NR-A2) has been developed for the treatment of allergic rhinitis using a unique combination of extracts from seven medicinal plants including Phyllanthus emblica, Terminalia chebula, Terminalia bellerica, Albizia lebbeck, Piper nigrum, Zingiber officinale and Piper longum. In this study, the antioxidant efficacy of Aller-7 was investigated by various assays including hydroxyl radical scavenging assay, superoxide anion scavenging assay, 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2-azinobis-ethyl-benzothiozoline-sulphonic acid diammonium salt (ABTS) radical scavenging assays. The protective effect of Aller-7 on free radical-induced lysis of red blood cells and inhibition of nitric oxide release by Aller-7 in lipopolysaccharide-stimulated murine macrophages were determined. Aller-7 exhibited concentration-dependent scavenging activities toward biochemically generated hydroxyl radicals (IC50 741.73 microg/ml); superoxide anion (IC50 24.65 microg/ml by phenazine methosulfate-nicotinamide adenine dinucleotide [PMS-NADH] assay and IC50 4.27 microg/ml by riboflavin/nitroblue tetrazolium [NBT] light assay), nitric oxide (IC50 16.34 microg/ml); 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical (IC50 5.62 microg/ml); and 2,2-azinobis-ethyl-benzothiozoline-sulphonic acid diammonium salt (ABTS) radical (IC50 7.35 microg/ml). Aller-7 inhibited free radical-induced hemolysis in the concentration range of 20-80 microg/ml. Aller-7 also significantly inhibited nitric oxide release from lipopolysaccharide-stimulated murine macrophages. These results demonstrate that Aller-7 is a potent scavenger of free radicals and that it may serve.     

Antioxidative Activity of the Hydrolytic Enzyme Treated Sorbus commixta Hedl. and its Inhibitory Effect on Matrix Metallopro- teinase- 1 in UV Irradiated Human Dermal Fibroblasts

Superoxide radical scavenging activity and DPPH radical scavenging activity were assessed in order to evaluate the antioxidant effect of the Sorbus commixta Hedl. extract (SCoE). SCoE was also treated with several carbohydrate-hydrolytic enzymes that significantly increased the total phenol and flavonoid composition of SCoE. The enzymatically treated SCoE was then assessed for antioxidative activity. The most efficient radical scavenging activity was observed when SCoE was treated with -glucanase. The radical scavenging activity of beta-glucanase-treated SCoE (beta-GSCoE) enhanced the viability of human dermal fibroblasts (HDFs) exposed to ultraviolet (UV) light. The intracellular reactive oxygen species (ROS) scavenging activity of beta-GSCoE was assessed using UVB (20 mJ/cm2)-irradiated HDFs. UVB irradiation increased dichlorofluorescein (DCF) fluorescence, which was measured by a 5-(6-)chloromethyl-2',7'- dichlorodihydrofluorescein diacetate (CM-H2DCFDA). DCF-fluorescence was significantly decreased in the beta-GSCoE-containing culture medium, suggesting that beta-GSCoE scavenges free radicals. The protective effect was further verified by assessing the expression of matrix metalloproteinase-1 (MMP-1) in UVA-irradiated HDFs. The treatment of UVA-irradiated HDFs with beta-GSCoE resulted in a dose-dependent decrease in the expression level of MMP-1 protein and mRNA. These results suggest that beta-GSCoE may mitigate the effects of photoaging in skin by reducing UV-induced adverse skin reactions.     

Antioxidant constituents and a new triterpenoid glycoside from<Emphasis Type="Italic">Flos Lonicerae</Emphasis>

As a component of our continuing investigations into herb-derived antioxidant agents, we have evaluated the antioxidant effects of Flos Lonicerae (Lonicera japonica flowers), via 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, total reactive oxygen species (ROS), hydroxyl radical (*OH), and peroxynitrite (ONOO-) assays. Among the methanolic extract and the dichloromethane, ethyl acetate, n-butanol, and water fractions, the EtOAc fraction of Flos Lonicerae exhibited marked scavenging/inhibitory activities, as follows: IC50 values of 4.37, 27.58 +/- 0.71, 0.47 +/- 0.05, and 12.13 +/- 0.79 microg/mL in the DPPH, total ROS, ONOO-, and *OH assays, respectively. Via a bioactivity-guided fractionation approach, a new triterpenoid glycoside, oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-xylopyranosyl(1-->6)]-beta-D-glucopyranosyl ester (12), along with eleven known compounds, including chrysoeriol (1), luteolin (2), 5-hydroxymethyl-2-furfural (3), caffeic acid (4), protocatechuic acid (5), chrysoeriol 7-O-beta-D-glucopyranoside (6), isorhamnetin 3-O-beta-D-glucopyranoside (7), kaempferol 3-O-beta-D-glucopyranoside (8), quercetin 3-O-beta-D-glucopyranoside (9), hederagenin 3-O-alpha-L-arabinopyranoside (10), and luteolin 7-O-beta-D-glucopyranoside (11), were isolated from the EtOAc fraction. The structures of isolated compounds 1-12 were elucidated via spectroscopic analyses. Compound 12 was isolated from a natural source for the first time. Compounds 2, 4, 5, 7, 9, and 11 evidenced marked scavenging activities, with IC50 values of 2.08-11.76 microM for DPPH radicals, and 1.47-6.98 microM for ONOO-.     

5,7,4’-三羟基异黄酮基-8-对偶氮苯磺酸的合成及其抗氧化活性研究

目的合成未见文献报道的化合物5,7,4’-三羟基异黄酮基-8-对偶氮苯磺酸,并探讨其抗氧化活性。方法采用IR、1H-NMR、元素分析对其结构进行表征。利用荧光光谱法研究目标物对羟基自由基的清除活性,采用紫外光谱法研究目标物对超氧阴离子自由基和1,1-二苯基-2-苦肼基自由基(DPPH·)的清除活性。结果与结论目标化合物对1,1-二苯基-2-苦肼基自由基、羟基自由基和超氧阴离子自由基都有良好的清除活性,而且对羟基自由基的清除活性远高于对超氧阴离子自由基和1,1-二苯基-2-苦肼基自由基的清除活性。     

Antioxidant activity of caffeic acid (3,4-dihydroxycinnamic acid)

Caffeic acid (3,4-dihydroxycinnamic acid) is among the major hydroxycinnamic acids present in wine; sinapic acid, which is a potent antioxidant. It has also been identified as one of the active antioxidant. In the present study, the antioxidant properties of the caffeic acid were evaluated by using different in vitro antioxidant assays such as 2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH) scavenging, total antioxidant activity by ferric thiocyanate method, total reductive capability using the potassium ferricyanide reduction method, superoxide anion radical scavenging and metal chelating activities. alpha-Tocopherol, trolox, a water-soluble analogue of tocopherol, butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) were used as the reference antioxidant compounds. At the concentrations of 10 and 30 microg/mL, caffeic acid showed 68.2 and 75.8% inhibition on lipid peroxidation of linoleic acid emulsion, respectively. On the other hand, 20 microg/mL of standard antioxidant such as BHA, BHT, alpha-tocopherol and trolox indicated an inhibition of 74.4, 71.2, 54.7 and 20.1% on peroxidation of linoleic acid emulsion, respectively. In addition, caffeic acid is an effective ABTS(+) scavenging, DPPH scavenging, superoxide anion radical scavenging, total reducing power and metal chelating on ferrous ions activities.     

Flavonoids from the flower ofRhododendron yedoense var.Poukhanense and their antioxidant activities

Antioxidant flavonoids have been isolated from the flower of Rhododendron yedoense var. poukhanense. One new flavonoid and three known flavonoids, quercetin-5-O-beta-D-glucopyranoside (1), quercetin (3), and quercitrin (4), were isolated from the butanol and ethyl acetate extracts of the plant. The new flavonoid was identified as myricitrin-5-methyl ether (2). The isolation of these flavonoids from this plant, for the first time, is a valuable finding. The flavonoids were evaluated for their antioxidant activities using 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH), TBARS (thiobarbituric acid reactive substance) and superoxide anion radical (O2-) in the xanthine/xanthine oxidase assay system. In the DPPH scavenging assay, the IC50 values were 4.5 +/- 0.48 microM for compound 2 and 9.7 +/- 0.29 microM for compound 3, which showed an antioxidant activity approximately 1.5-2 times higher than the antioxidant activity of alpha-tocopherol (9.8 +/- 0.94 microM). Additionally, the antioxidant activities of myricitrin-5-methyl ether (2) (IC50 = 1.7 +/- 0.22 microM) and quercetrin (4) (IC50 = 1.9 +/- 0.63 microM) were higher than that of L-ascorbic acid (IC50 = 7.4 +/- 0.63 microM) when evaluated using a TBARS assay. Compound 2 showed high activity in both the inhibition of xanthine oxidase (1.1 +/- 0.21 mM) and in the activation of superoxide scavenging.     

Assessment of the antioxidant activities of Brazilian extracts of propolis alone and in topical pharmaceutical formulations

The antioxidant activity of extracts of propolis and of formulations added with these extracts were measured by scavenging different radicals in different systems. For the ethanolic extract of propolis (EEP) and the glycolic extract of propolis (GEP) the IC50 observed were respectively of 0.024 and 0.035 microL/mL in scavenging hydroxyl radical, 0.016 and 0.012 microL/mL in inhibiting lipid peroxidation, 0.22 and 0.24 microL/mL in inhibiting chemiluminescence produced in the H2O2/luminol/horseradish peroxide (HRP) system and about 0.005 microL/mL for both extracts in inhibiting chemiluminescence produced in the xanthine/luminol/xanthine oxidase (XOD) system. The antioxidant activity of extracts of propolis in the formulations was not able to be assessed neither using the deoxyribose assay, since the formulation components interfered in the assay measurements, nor using chemiluminescence in the H2O2/luminol/HRP system, since this method did not show to be sensitive for the extract of propolis evaluation. However, the antioxidant activity of extracts of propolis could be successfully evaluated in the formulations using both lipid peroxidation and chemiluminescence generated in the xanthine/luminol/XOD system inhibitions.     

Antioxidant caffeic acid derivatives from leaves of<Emphasis Type="Italic">parthenocissus tricuspidata</Emphasis>

Five caffeic acid derivatives; methyl ester of caffeoylglycolic acid (1), dimethyl ester of caffeoyltartaric acid (2), dimethyl ester of caffeoyltartronic acid (3), monomethyl ester of caffeoyltartronic acid (4), methyl ester of caffeic acid (5), and some other secondary metabolites including; quercetin, quercetin 3-O-beta-D-glucuronide methyl ester, kaempferol, 3,5,7,4'-O-tetramethylkaempferol, beta-sitosterol glucoside, 2alpha-hydroxyursolic acid and 2,24-dihydroxyursolic acid, have been isolated and characterized. All the isolated compounds were characterized with the help of NMR spectroscopy and mass spectrometry. Structure of compound 3 was also confirmed by a single X-ray crystallographic technique. Isolates were evaluated for antioxidant activities and most of the tested compounds were found to be potent in DPPH free radical scavenging (IC50 = 4.56-14.17 microg/mL) and superoxide anion scavenging (IC50 = 0.58-7.39 microg/mL) assays.     

Cytotoxic and antioxidant activity of Achillea alexandri-regis

The cytotoxicity and antioxidant properties of herb extracts of Achillea alexandri-regis were studied. Combined chloroform and ethylacetate extracts exhibited a pronounced cytotoxic effect against HeLa cancer cells (IC50 = 25.92 +/- 4.96 microg/ml), and lower cytotoxicity against K562 leukemia cells (IC50 = 48.59 +/- 18.31 microg/ml). The methanol extract was found to be a moderately cytotoxic in vitro agent against HeLa and K562 cells. No suppressive activity was detected on non-malignant peripheral blood mononuclear cells (PBMC). The antioxidant activity of the methanol extract was assessed by DPPH radical scavenging. The methanol extract of A. alexandri-regis showed concentration dependent DPPH radical scavenging activity with IC50 = 36.14 +/- 0.05 microg/ml.     

Antioxidant and free radical-scavenging activity of Choto-san and its related constituents

The antioxidant properties of Choto-san and its related constituents such as Chotoko and Choto-san without Chotoko, and phenolic compounds contained in Chotoko such as epicatechin, caffeic, acid and quercetin were evaluated. In the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging assay, the scavenging activity of Chotoko (IC(50) 14.3 microg/ml) was found to be higher than that of Choto-san (IC(50) 206.2 microg/ml) and Choto-san without Chotoko (IC(50) 244.3 microg/ml). Epicatechin (IC(50) 10.4 microM), caffeic acid (IC(50) 13.8 microM), and quercetin (IC(50) 7.1 microM) also revealed scavenging activity against DPPH radicals. Choto-san (IC(50) 67.7 microg/ml) exhibited stronger inhibitory activity against superoxide anion formation than Choto-san without Chotoko (IC(50) 92.4 microg/ml) but weaker activity than Chotoko (IC(50) 18.3 microg/ml). The generation of superoxide anion was also inhibited by epicatechin (IC(50) 175.2 microM), caffeic acid (IC(50) 141.7 microM), and quercetin (IC(50) 18.7 microM). In a hydroxyl radical-scavenging experiment, Choto-san (IC(50) 2.4 mg/ml), Chotoko (IC(50) 2.2 mg/ml), Choto-san without Chotoko (IC(50) 2.8 mg/ml), epicatechin (IC(50) 3.9 mM), caffeic acid (IC(50) 3.6 mM), and quercetin (IC(50) 1.9 mM) exhibited activity. In NG108-15 cells, when added simultaneously with H(2)O(2) (500 microM), Choto-san (250 microg/ml), Chotoko (250 microg/ml), Choto-san without Chotoko (500 microg/ml), epicatechin (200 microM), caffeic acid (200 microM), and quercetin (200 microM) effectively protected cells from oxidative damage. In conclusion, the present results provide evidence that Choto-san acts as an antioxidant and cytoprotective agent against oxidative damage, which is due at least partly to the phenolic compounds contained in Chotoko.     

Free radical scavenging activity of red ginseng aqueous extracts

This study was performed to investigate the free radical scavenging activity of Panax red ginseng C.A. Meyer aqueous extract on 1,1-dipheny-2-picrylhydrazyl (DPPH), carbon-centered radical, hydroxyl and superoxide radicals using Electron Spin Resonance (ESR) spectrometer and spin-trapping techniques. Two different Red ginseng aqueous extracts prepared by boiling water or room temperature extraction exhibited no significant difference in free radical scavenging activity. Ginseng extracts completely eliminated DPPH radical at 2 mg/ml. About 0.5 mg/ml ginseng extracts quenched 80% carbon-centered free radicals generated from 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH). Hydroxyl radical and superoxide radical were generated by UV irradiation and trapped by 5,5-dimethyl-l-pyrroline-N oxide (DMPO). Ginseng extracts scavenged 40% of hydroxyl radical at 0.1 mg/ml. Two mg/ml ginseng extracts completely scavenged superoxide radical. Ginseng extracts did not scavenge nitric oxide. The ESR data demonstrate that red ginseng aqueous extract is not a strong free radical scavenger.     

A Prolyl endopeptidase-lnhibiting antioxidant fromPhyllanthus ussurensis

A prolyl endopeptidase inhibitor was isolated from the ethyl acetate soluble fraction of Phyllanthus ussurensis. The active compound was identified as an ellagitannin, corilagin. It was shown to non-competitively inhibit prolyl endopeptidase (PEP) with the IC50 value of 1.17x10(-6) microM. The Ki value was 6.70x10(-7) M. Corilagin was less inhibitory to other serine proteases such as chymotrypsin, trypsin, and elastase, indicating that it was relatively a specific inhibitor of PEP. Corilagin also effectively inhibited reactive oxygen species such as hydroxide and superoxide anion radical, hydrogen peroxide, and DPPH. Especially, corilagin showed potent scavenging activity on the superoxide anion radical in the ESR method (IC50 = 3.79x10(-6) M) as well as xanthine oxidase system.