• Korean Journal of Pharmacognosy
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• v.45 no.4
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• pp.300-314
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• 2014

An ultra-performance liquid chromatography-electrospray ionization-mass spectrometer (UPLC-ESI-MS) method was established for the simultaneous quantification of eighteen marker compounds in traditional Korean formula, Cheonwangbosimdan (CWBSD). In addition, we evaluated the antioxidant effects of CWBSD. Eighteen marker components were separated on a UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) and kept at $45^{\circ}C$ by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate was 0.3 mL/min and the injection volume was $2.0{\mu}L$. The antioxidant activities of CWBSD were assessed by measuring free radical scavenging activities on 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The calibration curves of all analytes showed good linearity (correlation coefficient ${\geq}0.9937$) within the test ranges. The limits of detection and quantification for the 18 marker compounds were 0.01-4.71 ng/mL and 0.03-14.13 ng/mL, respectively. The contents of the 18 compounds in CWBSD extract ranged from none to $1701.00{\mu}g/g$. The CWBSD showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were $149.42{\mu}g/mL$ and $339.24{\mu}g/mL$.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.383.3-384
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• 2002

Rosmarinic acid (1) and methyl rosmarinic acid (2), isolated from the ethyl acetate soluble fraction of the methanolic extract of Salvia miltiorrhiza Bunge (Lamiaceae) were found to be the tyrosinase inhibitors and scavengers of 1, 1-diphenyl-2-picrylhydrzyl (DPPH) radical. Compounds 1 and 2 inhibited the oxidation of L-tyrosine catalyzed by mushroom tyrosinase with $IC_{50}$/ of 16.8 $\mu\textrm{M}$ and 21.5 $\mu\textrm{M}$. respectively. It compared well with kojic acid. a well-known tyrosinase inhibitor. with an $IC_{50}$ of 22.4 $\mu\textrm{M}$. The inhibitory kinetics, analyzed by a Lineweaver-Burk plot, found rosmarinic acid and its methyl ester to be competitive inhibitors with $K_{i}$ of $2.35{\times}10^{-5}M$ and $1.52{\times}10^{-5}M$ respectively. In addition, compounds 1 and 2 showed the scavenging activities on DPPH radical, with $IC_{50}$ of 4.27 $\mu\textrm{M}$ and 3.05 $\mu\textrm{M}$. respectively. These scavenging effects were more potent than that of L-ascorbic acid ($IC_{50}$ = 11.75$\mu\textrm{M}$).

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.4
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• pp.275-286
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• 2008

In this study, we investigated the anti-oxidative, anti-wrinkle and whitening effects of Platycarya strobilacea bark extracts. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract / fractions of Platycarya strobilacea was in the order: 50% ethanol extract ($6.75{\mu}g/mL$) < deglycosylated aglycone fraction ($6.62{\mu}g/mL$) < ethyl acetate fraction ($4.15{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Platycarya strobilacea extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was ethyl acetate fraction (OSC50, $0.56{\mu}g/mL$) < 50% ethanol extract ($0.02{\mu}g/mL$) < deglycosylated aglycone fraction ($0.01{\mu}g/mL$). The deglycosylated aglycone fraction showed the most prominent scavenging activity. The protective effects of extract / fractions of Platycarya strobilacea on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethanol extract (50%) suppressed photohemolysis in a concentration dependent manner, particularly ethyl acetate fraction exhibited the most prominent cellular protective effect (${\tau}_{50}$, 717.27 min at $10{\mu}g/mL$). The inhibitory effect of Platycarya strobilacea extracts on tyrosinase were investigated to assess their whitening efficacy. Finally, their anti-elastase activities were measured to predict the anti-wrinkle efficacy in the human skin. The inhibitory effect ($IC_{50}$) on tyrosinase of some Platycarya strobilacea extracts was 50% ethanol extract ($243.98{\mu}g/mL$) < ethyl acetate fraction ($153.87{\mu}g/mL$) < deglycosylated aglycone fraction ($137.53{\mu}g/mL$). Also, The inhibitory effect of elastase ($IC_{50}$) of some Platycarya strobilacea extracts was 50% ethanol extract ($31.01{\mu}g/mL$) < ethyl acetate fraction ($14.42{\mu}g/mL$) < deglycosylated aglycone fraction ($1.48{\mu}g/mL$). The cream containing the ethyl acetate fraction of Platycarya strobilacea extracts was formulated. The skin hydration, transepidermal water loss, and the whitening effects were investigated after topical application of the cream. The skin hydration of cream containing extract was increased by $2{\sim}8%$ than the placebo cream, transepidermal water loss was decreased. The cream containing extract suppressed the melanogenesis of skin by 9.55% than the placebo cream. These results indicate that extract / fractions of Platycarya strobilacea can function as antioxidants in biological systems, particularly skin exposed to UV radiation by anti-oxidative activity and protect cellular membranes against ROS. The inhibitory effect on elastase and tyrosinase, and the increase of skin hydration and the whitening effect of the cream containing extract could be applicable to new functional cosmetics for antiaging.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.4
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• pp.357-363
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• 2015

The purpose of this study was to compare the inhibitory effect of bevacizumab on human Tenon's fibroblasts (HTFs) cultured from primary and recurrent pterygium. Cultured HTFs were exposed to 2.0, 5.0, 7.5, and 15.0 mg/mL concentration of bevacizumab for 24 hours. The 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase leakage assays were then performed to assess fibroblast metabolism and viability. The matrix metalloproteinase (MMP), procollagen type I C terminal propeptide (PIP), and laminin immunoassays were performed to examine extracellular matrix production. Changes in cellular morphology were examined by phase-contrast and transmission electron microscopy. Both metabolic activity and viability of primary and recurrent pterygium HTFs were inhibited by bevacizumab in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. Both types of HTFs had significant decreases in MMP-1, PIP, and laminin levels. Distinctly, the inhibitory effect of bevacizumab on MMP-1 level related with collagenase in primary pterygium HTFs was significantly higher than that of recurrent pterygium. Significant changes in cellular density and morphology both occurred at bevacizumab concentrations greater than 7.5 mg/mL. Only primary pterygium HTFs had a reduction in cellular density at a bevacizumab concentration of 5.0 mg/mL. Bevacizumab inhibits primary and recurrent pterygium HTFs in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. As the primary HTFs produces larger amounts of MMP-1 compared to recurrent HTFs, significant reduction in MMP-1 level in primary pterygium HTFs after exposure to bevacizumab is likely to be related to the faster cellular density changes in primary pterygium HTFs.

• Food Science and Biotechnology
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• v.15 no.3
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• pp.482-484
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• 2006

The antioxidant activity of Bacillus polyfermenticus SCD was studied by partially purified culture extracts using various methods: ammonium sulfate precipitation, adsorption to Diaion HP-20 columns using polar solvents, and extraction using non-polar solvents. The 1,1-diphenyl-2-picyrylhydrazyl (DPPH) radical scavenging activity of these partially purified fractions was then investigated. The precipitate isolated using 75%-saturated ammonium sulfate was shown to contain about 77.2% DPPH radical scavenging activity. Using the Diaion HP-20 resin adsorption method, the fraction obtained using 60% ethanol and 60% methanol possessed 76.7 and 89.5% DPPH radical scavenging activity, respectively. Fractions obtained by extracting with the non-polar solvents 80 mg/mL chloroform, 80 mg/mL n-hexane, 80 mg/mL ethyl acetate, and 80 mg/mL butanol contained 68.4, 75.0, 70.7, and 87.5% DPPH radical scavenging activity, respectively. Further study is needed to characterize the antioxidant substance(s) released by B. polyfermenticus SCD cultures.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.39 no.4
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• pp.295-302
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• 2013

In this study, the antioxidative effects and inhibitory activities of unripened fruit extract of Rubus coreanus Miquel (R. coreanus Miquel) on tyrosinase were investigated and the potential applicability as a cosmeceutical ingredients was evaluated. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of unripened fruit extract of R. coreanus Miquel. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activites ($FSC_{50}$) of 50% ethanol extract (6.56 ${\mu}g/mL$) and ethyl acetate fraction (6.14 ${\mu}g/mL$) of unripened fruit extract of R. coreanus Miquel were higher than (+)-${\alpha}$-tocopherol (8.98 ${\mu}g/mL$), which is known as a typical hydrophobic antioxidant. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of 50% ethanol extract (0.83 ${\mu}g/mL$), ethyl acetate fraction (0.84 ${\mu}g/mL$) and aglycone fraction (1.13 ${\mu}g/mL$) of R. coreanus Miquel on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were higher than L-ascorbic acid (1.5 ${\mu}g/mL$), which is known as s typical hydrophilic antioxidant. The cellular protective effect of extract and fraction of unripened fruit extract of R. coreanus Miquel on the rose bengal sensitized photohemolysis of human erythrocytes was increased in a concentration dependent manner (1 ~ 50 ${\mu}g/mL$). And 50% ethanol extract in 50 ${\mu}g/mL$ showed the most protective effect among extracts (${\tau}_{50}$ = 296.3 min). The inhibitory effects on tyrosinase of ethyl acetate and agylcone fractions were higher than arbutin. These results indicate that unripened fruit extracts of R. coreanus Miquel can be applied to antioxidant scavenging ROS including radical as an alternative whitening agent to replace arbutin.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.2
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• pp.191-198
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• 2011

In this study, antioxidative effects and inhibitory effects of Geum aleppicum Jacq. extracts on tyrosinase and elastase were investigated. The ethyl acetate fraction of G. aleppicum Jacq. extract ($4.70\;{\mu}g$/mL) showed the most prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity (FSC50). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some G. aleppicum Jacq. extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction showed the most prominent ROS scavenging activity ($0.22 \;{\mu}g$/mL). The protective effects of extract/fraction of G. aleppicum Jacq. against the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The G. aleppicum Jacq. extracts suppressed photohemolysis in a concentration dependent manner ($1{\sim}25{\mu}g$/mL), particularly the ethyl acetate fraction exhibited the most prominent celluar protective effect (${\tau}_{50}$, 416.20 min at $10 \;{\mu}g$/mL). The inhibitory effect of G. aleppicum Jacq. extracts on tyrosinase and elastase were investigated to assess their whitening and anti-winkle efficacy. The half maximal inhibitory concentration ($IC_{50}$) of the ethyl acetate fraction on tyrosinase was $95.23\;{\mu}g$/mL. The $IC_{50}$ of 50 % ethanol extract and the ethyl acetate fraction on elastase were $6.27 \;{\mu}g$/mL and $4.31 \;{\mu}g$/mL, respectively. These results indicate that extract/fraction of G. aleppicum Jacq. can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Especially the ethyl acetate fraction of G. aleppicum Jacq. extracts could be applicable to new functional cosmetics for antioxidant, antiaging.

• Archives of Pharmacal Research
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• v.27 no.6
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• pp.610-614
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• 2004

The in vitro antioxidant properties of some flavone-6(4)-carboxaldehyde oxime ether deriva-tives (Ia-f, lIa-f） were determined by their effects on the rat liver microsomal NADPH-dependent lipid peroxidation (LP） levels by measuring the formation of 2-thiobarbituric acid reactive substances. The free radical scavenging properties of the compounds were also examined in vitro by determining their capacity to scavenge superoxide anions and interact with the stable free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH). The most active compounds, lib (Flavone-4＇-carboxaldehyde-O-ethyl oxime） and Id (Flavone-6-carboxaldehyde-O-［2-(1-pyrolidino） ethyl］ oxime）, caused 98 and 79％ inhibition of superoxide anion production and DPPH stable free radical at $10^{-3}{\;}M$, respectively.

• Journal of Integrative Natural Science
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• v.7 no.3
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• pp.173-182
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• 2014

To simplify the different biological investigation of the methanolic extract and solvent-solvent partitioning of Lablab purpures (L. purpures) bark. In-vitro anti-oxidant study was determined using total DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing zone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and thrombolytic activity by clot disruption method. The antioxidant potential was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteau reagents using butylated hydroxytolune (BHT) and ascorbic acid as standards. The Aqueous soluble fraction revealed the highest free radical scavenging activity ($IC_{50}=48.76{\mu}g/mL$). The antimicrobial screening of the bark of L. purpures exhibited mild to moderate activity in test microorganisms. The CSF showed the maximum relative percentage inhibition against Salmonella parathyphi (34.2%) for bacteria and C. albicans (28.8%) for fungi whereas, lowest relative percentage inhibition against Sarcina lutea (22.0%) for bacteria and Aspergillus niger (24.4%) for fungi. In the brine shrimp lethality bioassay, The $LC_{50}$ values of Carbon tetrachloride and N-Hexane soluble fraction were found $92.18{\mu}g/mL$, and $68.95{\mu}g/mL$ respectively while the $LC_{50}$ values of standard Vincristine sulphate was $1.37{\mu}g/mL$. The methanolic extract and its organic soluble fractions of Lablab purpureus at concentration 2.0 mg/mL, significantly protected the lysis of erythrocyte membrane induced by hypotonic solution and heat as compared to the standard, acetyl salicylic acid (0.10 mg/mL). The MSF and AQSF produced 61.48 % and 53.75% inhibition of hemolysis of RBC caused by hypotonic solution respectively, whereas acetyl salicylic acid (0.10 mg/mL) showed 76.42%. Ethanol extract of L. purpures and all of its different partitions exhibited moderate thrombolytic activity of 37.25%-2.40%. Very good preliminary screening and simplified experiments were able to show the different biological activity of methanolic extract and its soluble fractions of L. purpures at a time.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.391-402
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• 2014

In this study, the antioxidative effects and component analysis for the extracts of Rumex acetosa L. were investigated. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from dried R. acetosa L.. Free radical scavenging activities (1,1-diphenyl-2- picrylhydrazyl) size of, in the order of aglycone fraction > ethyl acetate fraction > 50% ethanol extract, aglycone fraction ($45.10{\mu}g/mL$) showed the highest radical scavenging activity. Reactive oxygen species (ROS) scavenging activity (total antioxidant capacity, $OSC_{50}$) on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system was also, in the order of ethyl acetate fraction> aglycone fraction> 50% ethyl acetate fraction, ethyl acetate fraction ($2.68{\mu}g/mL$) was shown a great antioxidant capacity. The total antioxidant capacity of the ethyl acetate fraction was found to be greater than L-ascorbic acid, known as a typical hydrophilic antioxidant ($6.88 {\mu}g/mL$). The cellular protective effects of R. acetosa L. extracts on the $^1O_2$-induced cellular damage of human erythrocytes were exhibited at all concentration-dependent ($1{\sim}25{\mu}g/mL$). Especially, aglycone fraction (${\tau}_{50}$, 104.80 min) in $25{\mu}g/mL$ showed the most protective effect among extracts. Components of the ethyl acetate fraction obtained from R. acetosa L. extracts were analyzed by TLC, HPLC chromatogram, LC/ESI-MS/MS. As a result, the ethyl acetate fraction contained several flavonoids, such as orientin, isoorientin, vitexin, isovitexin. These results indicate that the R. acetosa L. extracts can be used as antioxidants in biological systems, particularly skins exposed to UV radiation by quenching and/or scavenging $^1O_2$ and other ROS. Thus, the extracts of R. acetosa L. could be applicable to new anti-aging cosmeceutical ingredients.