더덕(Codonopsis lanceolata) 추출물의 in vitro 및 in vivo 항산화 효과

This study was conducted to investigate the changes in saponin content and antioxidant activity of crude ginseng and extruded ginseng by using different solvent extraction methods. Each of the fractions was first extracted by 80% ethanol followed by ether treatment to remove the lipid components. Water soluble components were separated by ethylacetate and water saturated butanol. Four fraction, including 80% ethanol, ethylacetate, butanol and water were obtained from crude and extruded ginsengs to analyze saponin content and antioxidant activity. Saponin content and antioxidant capacity of each of the four fractions were measured by LC/MS analysis and ORAC(Oxygen Radical Absorbance Capacity) assay, respectively. It was found that a major portion of saponin was present in ethyl acetate and water saturated butanol fractions. When extracted by 80% ethanol, ginsenoside Rb1 and Rg1 were mostly found in crude ginseng, while ginsenoside Re and Rb1 were detected in extruded ginseng. Even though Rh1 and Rg3 were found in a very small quantity in crude ginseng, there was a significant quantity of both in extruded ginseng when extracted by 80% ethanol. Similar tendency was also observed in extruded ginseng fraction when extracted with ethyl acetate and butanol. In crude ginseng, the level of Rg1 was the highest among other ginsenosides upon extraction by ethyl acetate, while Rh1 and Rg3 were predominantly found by employing similar solvent extraction in the extruded ginseng. Also, Rg1, Re and Rb1 were also found in the extruded ginseng with small quantity. Rg1, Re and Rb1 were found in crude ginseng by butanol extraction, while Rb1 and Re were extracted from the extruded ginseng. Overall, there was no difference in the saponin content between crude ginseng and extruded ginseng when extracted by butanol and water, but twice as much of saponin was obtained by 80% ethanol extraction and 6 times more saponin were obtained in ethyl acetate fraction in the extruded ginseng. Antioxidant capacity of crude ginseng as determined by ORAC assay was higher in 80% ethanol(high in many different kinds of biological compounds) and water saturated butanol(high in polar saponin) fractions than the ethyl acetate and water fractions. No difference in antioxidant capacity was observed between crude and extruded ginseng. However, antioxidant capacity of ethyl acetate and water fractions in extruded ginseng was significantly higher than crude ginseng(P>0.05). All the fractions in both, crude and extruded ginseng possessed antioxidant capacity and even water fractions that contained almost no saponin had some antioxidant capacity. While determining correlation coefficient between fractions in extruded ginseng by Pearson correlation, it was observed that 80% ethanol fraction was in correlation with ethyl acetate(P>0.01) and ethanol(P>0.001) and in the case of ethylacetate, correlation was observed only with butanol fraction(P>0.05).Key words: solvent, saponin, extruded ginseng, antioxidant activity, oxygen radical absorbance capacity assay

Commercially available tea infusions are the major source of catechins for preparing bottled tea beverages and tea supplements available in the market today. In the present study, we analyzed five tea infusions to measure the total antioxidant capacity (TAC) by oxygen radical absorbance capacity (ORAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity (DRSC) assays, total polyphenol content by the colorimetric method and individual catechin content by high-performance liquid chromatography. Four major tea catechins were also analyzed for their TAC to reveal differential antioxidant behavior of the tea infusions, resulting in the ORAC and DRSC methods. The correlation coefficients between DRSC and the total polyphenol or total catechin content of the tea infusions were 1.0 and 0.99. However, the values fall to 0.73 and 0.69, respectively, while the ORAC activity was correlated with total polyphenol and total catechin content. Determining the TAC of individual tea catechins showed that ORAC of epicatechin was seven-fold higher than that of epigallocatechin gallate; on the contrary, epigallocatechin gallate showed significantly (P < 0.05) stronger DRSC activity than epicatechin. By evaluating the structure−activity relationship, this study further revealed that OH substitution at the 3′ position in pyrogallol moieties contributes to the lower ORAC value of epigallocatechin and epigallocatechin gallate comparing with their non-3′-OH counterparts, such as epicatechin and epicatechin gallate, respectively. Also, numbers of OH substitutions were poorly correlated with the observed ORAC value unlike the DRSC. Overall, results of this study enabled us to hypothesize that substances having a lower TAC value in the ORAC assay compared with that in DPPH assays may pertain to a pro-oxidant effect by generating reactive oxygen species in an aqueous buffer, at a physiological pH. We also propose that substances exhibiting lower TAC value in the ORAC assay compared with that in the DPPH assay are powerful pro-oxidants compared with the substances showing a higher TAC value in the ORAC assay than that in the DPPH assay.

Antioxidant capacity of extracts of different polarity obtained from two Hypericum L. species (H. juniperinum and H. mexicanum) was assessed by means of total polyphenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assay, ferric reducing antioxidant power (FRAP) assay and oxygen radical absorbance capacity (ORAC) assay. Their phenolic acid composition was also determined by HPLC. The ethyl acetate extract of H. juniperinum was the most active in the ABTS, FRAP and TPC assays with 10867.48 ?mol TEAC/g, 242.80 mg AAE/g and 491.08 mg GAE/g respectively. On the other hand, the methanol extract obtained from H. mexicanum appeared as the most active extract in the DPPH assay (3714.23 ?mol TEAC/g). Similarly, the butanol fraction coming from the methanolic extract of H. mexicanum showed the highest activity in the ORAC assay (12910.06 ?mol TEAC/g). HPLC analysis of the extracts revealed the presence of phenolic acid compounds, such as chlorogenic (50.09 mg/g) and p-coumaric acids (63.36 mg/g) in H. mexicanum and p-coumaric acid (8.45 mg/g) in H. juniperinum. A high correlation between antioxidant activity and total polyphenol content was established. Specifically, H. mexicanum exhibited the highest ORAC capacity, which may be associated with the high content of chlorogenic and p-coumaric acids present in medium to polar extracts. Our results constitute a significant contribution to the study of antioxidant activity and the determination of the phenolic acid profile in both species. The analysed extracts showed promising antioxidant activity that could be useful in the pharmaceutical, cosmetic and food industries.

The present study was to investigate the antioxidant activity in ethanol and water extracts of root and stem of Sargassum coreanum. Antioxidant activities were evaluated by total polyphenol contents, DPPH radical scavenging activity, chelating effect, reducing power, and rancimat method. Total polyphenol contents of ethanol and water extracts were 32.79 mg/g and 15.55 mg/g, respectively. Ethanol extract showed higher DPPH radical scavenging activity than water extract and similar activity to BHT. Reducing power of extracts was increased in a concentration-dependent manner and ethanol extract had more reducing power than water extract. Ethanol and water extracts have little chelating effect at all concentrations. Antioxidant index (AI) of ethanol extract measured by Rancimat was higher than that of water extract, but their AI was lower than that of BHT. These results indicate that ethanol extract of S. coreanum root and stem has more potent antioxidant activity than water extract through DPPH radical scavenging and reducing power, and could potentially be used as a good source of natural antioxidants.

In this study, we investigated antioxidative activity, antibacterial activity against pathogenic strains including methicillin resistant Staphylococcus aureus (MRSA), and tyrosinase inhibitory activity in 75% ethanol extract of Taraxacum coreanum and its fractions. The total polyphenol and flavonoid contents of the extract were 238.59mg/g and 33.18mg/g and the total polyphenol and flavonoid contents of the ethyl acetate fraction were 427.81mg/g and 148.90mg/g as the highest content of fractions. In DPPH radical scavenging ability, values of the ethyl acetate and butanol fraction were 38.40 and 82.28 , respectively. In antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and MRSA, the ethyl acetate fraction showed stronger antibacterial activity than other fractions and the extract. Especially, the ethyl acetate fraction was exhibited effective antibacterial activity against MRSA. In the cytotoxicity measurement by MTT assay, the extract and fractions were exhibited Raw 264.7 cell viabilities of 96.32~143.21% as nontoxic result in concentration of 5~100 . As a result, the ethyl acetate fraction of the 75% ethanol extract from T. coreanum could be applicable to functional materials for related fields.

In this study, different solvent extracts of skullcap (Scutellaria baicalensis) were assayed for their total phenol content (TPC), antioxidant activity [determined as DPPH radical scavenging activity, superoxide dismutase (SOD)-like activity, oxygen radical absorbance capacity (ORAC) assay, and comet assay], and α-glucosidaseinhibitory activity. The TPC of skullcap ranged from 9.06 mg/g gallic acid equivalents (GAE) for acetone extract (AE) to 91.8 mg/g GAE for methanol extract (ME). AE, which had a low TPC, exhibited the highest DPPH radical scavenging activity and SOD-like activity. TPC positively correlated with the ORAC assay (r=0.96, p<0.001). All skullcap extracts significantly reduced the hydrogen peroxide-induced DNA damage in human leukocytes. ME with a high TPC and ORAC value showed the highest α-glucosidase inhibition. The difference in the biological activities of the extracts may be due to the differences in their chemical structure or polarity. Therefore, the results obtained indicate that might be a potential source of compounds with health-protective effects. ME, in particular, might be a prospective therapeutic agent for diabetes.

The aim of this study was to investigate the antioxidant and antimicrobial activities of various solvent (acetone, ethyl acetate, and ethanol) extracts from Lentinus edodes. The antioxidant activities were evaluated by measuring total polyphenol and flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity. Total polyphenol content and ABTS radical scavenging activity were highest in ethanol extract. ABTS radical scavenging activity of ethanol extract showed the highest value (98.5%), which was similar to that of ascorbic acid (95.7%). The ethyl acetate extract from Lentinus edodes showed relatively high total flavonoid content and DPPH radical scavenging activity. Negative correlations were found between total polyphenol contents and DPPH radical scavenging activities in Lentinus edodes extracts. Antimicrobial activities of the extracts were determined against Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus, Escherichia coli, Pseudomonas aeruginosa, and Enterobacter cloacae by the disc diffusion method. The acetone and ethanol extracts showed moderate antimicrobial activities against almost all tested microorganisms except E. coli and S. aureus, respectively. The ethyl acetate extract showed a significant growth inhibition effect against E. coli, Ent. cloacae, and B. subtilis.

The content of insoluble bound phenolic acids in pearled barley was determined by an analytical system consisting of alkaline hydrolysis extraction, high-performance liquid chromatographic separation and electrochemical detection. Insoluble bound phenolic acids in five pearled cultivars and fifteen breeding lines of barley comprised ferulic acid (4.3-34.2mg/100 g dry matter), sinapic acid (0.025-0.445 mg/100 g), caffeic acid(0.002-0.016 mg/100 g). Soluble free polyphenols comprised procyanidins(12.2-80.3 mg/100 g), catechin (0.1-28.2 mg/100 g), and total pholyphenolcomprised 152.4-324.0 mg-gallic acid equivalents/100 g. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity for the barley samples ranged from 403 to 1501 μmol-Trolox equivalents/100 g whereas those of oxygen radical absorbance capacity (ORAC) ranged from 1050 to 3816 μmol-Trolox equivalents/100 g. High correlation (0.980, p<0.01) was found between the DPPH and ORAC assays. Total polyphenol contents positively correlated with the DPPH (0.875, p<0.01)and ORAC (0.881, p<0.01)assays. The correlation coefficient between insoluble bound phenolic acids and total polyphenol contents was higher than that between soluble free polyphenols and total poly-phenol contents. Taken together, the insoluble bound phenolic acids appear to greately contributed to the antioxidant activity in pearled barley.

Chemical composition, antioxidant activity and inhibitory capacity of α-amylase, α-glucosidase, lipase and non-enzymatic glycation, in vitro, of the leaves of Cassia bakeriana Craib

In the present study, we investigated biological activities of Rosa davurica Pall. leaves in order to evaluate thepossibility as a natural biomaterial. The 80% ethanol extract and its subsequent fractions of Rosa davurica Pall. leaves wereprepared using several solvents with different polarities. The extraction yield was 33.4, 36.6, 25.2, 18.7, 5.8 and 5.8% in ethanolextract, aqueous, butanol, ethyl acetate, n-hexane, chloroform fractions, respectively. The ethyl acetate fraction(661.38 ㎎/g), butanol fraction (396.68 ㎎/g) and 80% ethanolic extract (239.54 ㎎/g) has higher total polyphenol contentsthan other fractions. The antioxidant activity was detected in ethanolic extract, ethyl acetate and butanol fractions. Theethyl acetate fraction showed the highest levels of DPPH radical scavenging activity (IC50, 4.77 ㎍/㎖). Moreover, the ethylacetate fraction significantly inhibited production of NO in LPS-stimulated macrophage RAW 264.7 cells without cytotoxicity. These results indicate that 80% ethanol extract and its fractions of Rosa davurica Pall. leaf, especially ethyl acetate fraction,have the properties of anti-oxidant and anti-inflammation, suggesting leaf of Rosa davurica Pall. may be a candidate fornatural and functional materials.

During storage, the viability of sperm in a honey bee (Apis mellifera) queen’s spermatheca can be decreased by reactive oxygen species. We hypothesized that the expression of antioxidant genes would increase in queen spermathecae after mating. We measured queen morphometric characteristics and expression levels of seven antioxidant-encoding genes in virgin and mated queen spermathecae. We identified a 12% increase in body weight and a fourfold increase in ovary weight in mated queens. There was a twofold higher expression of catalase, thioredoxin 2, and thioredoxin reductase 1 in the spermathecae of mated vs. virgin queens. Expression of the other antioxidant genes (glutathione S-transferase D1, superoxide dismutase 1, vitellogenin, and glyoxalase domain-containing 4-like (GLOD4L) in spermathecae was not different between mated and virgin queens. In drone semen, expression of antioxidant genes was overall low compared to queens except for GLOD4L, which was equivalent to that in queen spermathecae. Increased expression of antioxidant genes may assist in maintaining sperm viability inside the spermathecae of mated queens.

As an effort to develop functional ingredients and to discover their antioxidant activity, the total polyphenol content, total flavonoid content, 2,2-diphenyl-1-pcrylhydrazyl (DPPH) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC), oxygen radical absorption capacity (ORAC), and ferric reducing antioxidant power (FRAP) of Daphniphyllum macropodum were measured using a methanol extract and various solvent fractions. Based on the result, the total polyphenol content was highest in an ethyl acetate fraction of 212.33±1.05 mg GAE/g where the total flavonoid content was 77.96±0.21 mg QE/g. The DPPH radical scavenging activity was highest in an ethyl acetate fraction (IC50 : 47.10±1.32 μg/mL). The ethyl acetate fraction showed the highest TEAC (1052.23±13.94 mM TE/g), ORAC (9308.71±480.69 mM TE/g), and FRAP (2064.76±42.31 mM FE/g) in antioxidant capacity value. The total polyphenol and flavonoid contents significantly correlated with the antioxidant activity of D. macropodum extracts and fractions. In addition, the survival rate of HaCaT and HepG2 cells at 200 μg/mL was higher than 70%. With these results, it was confirmed that the ethyl acetate fraction of D. macropodum has great antioxidant potential. Thus, it can be expected to be developed into a specific functional ingredient.

Context: Premature aging usually occurred due to free radicals reducing the skins’ physiological functions. Muntingia calabura, a plant containing rich antioxidants, has the potential to overcome this problem. Aims: To evaluate the antioxidant capacity of M. calabura in inhibiting the premature aging process, to be potentially developed into an antiaging active ingredient. Methods: The samples were extracted using ethanol 96%, and processed into n-hexane, ethyl acetate, and ethanol fractions, respectively. Total phenolic content was determined, followed by the evaluation of antioxidant capacity through DPPH, FRAP, and ABTS assay. Further, anti-elastase was conducted using human neutrophil elastase as a skin degradation enzyme, followed by an anti-collagenase test. Finally, normal cell proliferation was also evaluated via the MTT method measuring cell viability on HDFa cells. Results: As the results, ethanol extract, ethyl acetate fraction, and ethanol fraction showed a strong antioxidant effect, having great capacity reducing DPPH, ABTS radicals, and also iron reduction, in contrast to n-hexane fraction that exhibited only weak activity. The antioxidant trend capacities were found directly correlated to total phenolic contents. Furthermore, the ethyl acetate fraction was found to have optimum activity in inhibiting elastase and collagenase enzymes, showing a similar impact on cell viability. Conclusions: The ethyl acetate fraction from M. calabura exhibits the prospect for further development to support its effectiveness as an active ingredient in antiaging cosmetics.

We recently reported the improved oxygen radical absorbance capacity (ORAC) assay using fluorescein (FL) as the fluorescent probe. The current ORAC(FL) assay is limited in hydrophilic antioxidant due to the aqueous environment of the assay. Lipophilic antioxidants mainly include the vitamin E family and carotenoids, which play a critical role in biological defense systems. In this paper, we expanded the current ORAC(FL) assay to lipophilic antioxidants. Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water solubility enhancer for lipophilic antioxidants. Seven percent RMCD (w/v) in a 50% acetone-H(2)O mixture was found to sufficiently solubilize vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). This newly developed ORAC assay (abbbreviated ORAC(FL-LIPO)) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrate that the ORAC(FL-LIPO) assay is reliable and robust. For the first time, by using 6-hydroxy-2,5,7,8-tetramethyl-2-carboxylic acid as a standard (1.0), the ORAC values of alpha-tocopherol, (+)-gamma-tocopherol, (+)-delta-tocopherol, alpha-tocopherol acetate, tocotrienols, 2,6-di-tert-butyl-4-methylphenol, and gamma-oryzanol were determined to be 0.5 +/- 0.02, 0.74 +/- 0.03, 1.36 +/- 0.14, 0.00, 0.91 +/- 0.04, 0.16 +/- 0.01, and 3.00 +/- 0.26, respectively. The structural information of oxidized alpha-tocopherol obtained by liquid chromatography/mass spectrometry reveals that the mechanism for the reaction between the vitamin E and the peroxyl radical follows the hydrogen atom transfer mechanism, which is in agreement with the notion that vitamin E is the chain-breaking antioxidant.

Background: Andrographis paniculata is a herbaceous plant in the Acanthaceae family, that is widely used as a traditional medicine in Asian countries and known to exhibit a wide range of pharmacological effects. Recent studies have provided an overview of the great potential of A. paniculata as an analgesic. The ethanol extract and ethyl acetate (EA) fraction of A.paniculata were shown to contain diterpene lactone compounds, which may be useful as a potential active ingredient in analgesic drugs. The development of a herbal medicine based drug requires an effective and high quality active ingredient. Therefore, this research was aimed to compare the analgesic activity of ethanol extract and EA fraction based on their andrographolide content and further to determine the more viable active substance for analgesic herbal medicine based drug development. Method: The andrographolide content in the ethanol extract and EA fraction was determined by High Pressure Liquid Chromatography (HPLC). Measurement of analgesic activity was performed by writhing test. The experimental animals were randomly divided into eight groups consisting of 5 mice in each. Group 1 (negative control) received 1% Tween-80 in normal saline. Group 2 (positive control) received a standard analgesic drug (diclofenac sodium) at a dose of 40 mg/kg body weight. Group 3, 4, and 5 received ethanol extract while Group 6, 7, and 8 received EA fraction, each at a dose of 12.5, 25, and 50 mg andrographolide/kg body weight, respectively. Each mouse was injected intraperitoneally with 1% acetic acid at a dose of 10 ml/kg body weight 30 minutes after oral administration of the treatments. The number of writhes were counted 5 min after acetic acid injection over a period of 45 min. Results: Andrographolide content in ethanol extract and EA fraction was 15.66±0.28 and 21.25±1.08 % w/w, respectively. Ethanol extract and EA fraction displayed analgesic activity of 67.68% and 70.91% respectively, at a dose of 50 mg andrographolide/kg body weight. The positive control at a dose of 40 mg/kg body weight showed an analgesic activity of 74.33%. Statistical analysis showed no significant differences between EA fraction at a dose of 50 mg andrographolide/kg body weight and ethanol extract at the same dose as well as the positive control (P&gt; 0.05). The effective dose 50% (ED50) of the ethanol extract and EA fraction was determined to be 29.49 and 25.55 mg/kg body weight, respectively. Conclusion: It was possible to use andrographolide content as an indicator for the analgesic activity of A.paniculata. Ethanol extract and EA fraction of A. paniculata at the same dose of andrographolide showed similar analgesic activity. The amount of ethanol extract which needed to reach similar analgesic activity was higher than EA fraction. Therefore, EA fraction likely has greater potential as an analgesic active substance due to its higher content of andrographolide; however further study is needed to develop it as a dosage form.