• Journal of Life Science
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• v.27 no.2
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• pp.225-232
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• 2017

This study aimed to evaluate several biological activities of Pharbitis nil and to isolate an anticancer agent from its methanol extract. Pharbitis nil seeds were extracted with methanol (PNM). Then, PNM was fractionated into solvent layers such as ethyl acetate fraction (PNE), butanol fraction (PNB), and water fraction (PNW). The biological activities of the fractions were analyzed for tyrosinase inhibition, lipase inhibition, DPPH-free radical scavenging, and cell growth inhibition. PNM showed strong growth inhibition of prostate cancer PC-3 cells. PNM was subjected to Diaion HP-20 and eluted stepwise with 50%, 80%, and 100% methanol. Then, for activity-guided fraction, each fraction was analyzed for growth inhibition of prostate cancer PC-3 cells by using an MTT assay. Because the 100% fraction showed significantly strong inhibitory activity, the fraction was further separated in the reverse phase C18, which was eluted with 80% and 90% methanol. The 90% fraction was further subjected to Sephadex LH-20 using a mobile solvent of 100% methanol. Finally, the compound PN was partially purified for HPLC analysis. PN showed cell growth inhibitory activity and induced the apoptosis and cell cycle arrest of prostate cancer PC-3 cells, as measured by flow cytometry. The results together suggest that Pharbitis nil possesses various biological activities, especially the inhibitory activity for the proliferation of prostate cancer PC-3 cells, suggesting the possibility of its use as an anticancer agent.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.36 no.4
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• pp.303-314
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• 2010

In this study, the antibacterial, antioxidative effect and component analysis of Pinus koraiensis leaf extracts were investigated. MIC values of the ethyl acetate fraction from P. koraiensis leaf extracts on P. acnes, S. aureus, P. ovale, and E. coli were 0.06 %, 0.25 %, 0.13 % and 0.50 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction on P. acnes, P. ovale. and S. aureus was more prominent. The aglycone fraction of P. koraiensis leaf extracts ($22.93\;{\mu}g/mL$) showed more higher free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. koraiensis leaf extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The 50 % ethanol extract ($0.70\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. Also the ethyl acetate ($1.04\;{\mu}g/mL$) and the aglycone fraction ($1.43\;{\mu}g/mL$) showed very high antoxidant activity. The protective effects of extract/fractions of P. koraiensis leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. koraiensis leaf extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). TLC and HPLC chromatogram of the ethyl acetate fraction obtained from hydrolysis of P. koraiensis leaf extracts revealed 2 main bands (PK-4, PK-6) and peaks (peak 1, peak 2), which were identified as kaempferol-3-O-glucoside (PK-6, peak 1) and kaempferol-3-O-arabinoside (PK-4, peak 2) by LC/ESI-MS/MS, respectively. These results indicate that extract/fractions of P. koraiensis can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membrane against ROS. Extract/fractions of P. koraiensis can be applicable to new cosmeceuticals for antioxidant, antiaging, and antibacterial activity.

• Journal of the Korean Applied Science and Technology
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• v.33 no.4
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• pp.647-657
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• 2016

In this study, the antioxidative effects and active component analysis of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Prunella vulgaris L. were investigated. The free radical scavenging activities ($FSC_{50}$) was investigated at 50% ethanol extract ($15.25{\mu}g/mL$), ethyl acetate fraction ($8.68{\mu}g/mL$), and aglycone fraction ($8.25{\mu}g/mL$) respectively. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay was investigated at 50% ethanol extract ($4.68{\mu}g/mL$), ethyl acetate fraction ($1.00{\mu}g/mL$), and aglycone fraction($1.02{\mu}g/mL$) respectively. In the cellular protective effect against $^1O_2$ induced cellular damage of human erythrocytes, extract/fractions of P. vulgaris L. were increased in a concentration dependent manner($1{\sim}25{\mu}g/mL$). Especially, ${\tau}_{50}$ of aglycone fraction at concentrations of $25{\mu}g/mL$ showed the most protective effects at 337.9 min. It's showed nine times higher (+)-${\alpha}$-tocopherol (${\tau}_{50}=38.7min$) as typical antioxidant in the $^1O_2$-induced photohemolysis of human erythrocytes. TLC and HPLC were used to analyse active components in the ethyl acetate fraction and aglycone fraction of P. vulgaris L. In ethyl acetate fraction, caffeic acid, rosmarinic acid, quercetin 3-${\beta}$-D-glucoside, rutin, kaempferol-3-O-rutinoside, astragalin (kaempferol-3-O-glucoside) were identified. In aglycone fraction, caffeic acid, rosmarinic acid, quercetin, kaempferol were identified. These results indicated that extract/fraction of P. vulgaris L. is may be used in cosmetics industry as natural antioxidants by quenching and/or scavenging $^1O_2$ and other ROS, and protecting cellular membranes.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.2
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• pp.125-134
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• 2009

In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component of non-fermented and fermented Lavandula angustifolia extracts were investigated. The ethyl acetate fraction of fermented extract (5.95 ${\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of L. angustifolia extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract (1.45 ${\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of L. angustifolia on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The L. angustifolia extracts suppressed photohemolysis in a concentration dependent manner (1 ${\sim}$ 50 ${\mu}g/mL$). The inhibitory effect of L. angustifolia extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase were determined with ethyl acetate fraction of L. angustifolia extract (144.80 ${\mu}g/mL$) and ethyl acetate fraction of fermented extract (122.40 ${\mu}g/mL$). Fractions of ethyl acetate and fermented extracts showed both 3 band in TLC and 3 peaks, 2 peaks in HPLC (340 nm), respectively. In each chromatography, fractions of ethyl acetate both from non-fermented and fermented L. angusfifolia have rosmarinic acid in common. These results indicate that the component and content of non-fermented and fermented extracts of L. angustifolia are different. Both of the extract of L. angustifolia can be used as an antioxidant.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.2
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• pp.143-152
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• 2011

In this study, the antioxidative activity, inhibitory effects on tyrosinase and elastase, and active components of Quercus salicina Blume extracts were investigated. Q. salicina Blume was extracted using 50 % ethanol, from which ethyl acetate and aglycone fractions were prepared. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity ($FSC_{50}$) of Q. salicina Blume aglycone fraction was the highest ($8.25\;{\mu}g$/mL). The luminol-dependent chemiluminescence assay showed that reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of Q. salicina Blume aglycone fraction on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system was the most prominent at $0.039\;{\mu}g$/mL. The protective effects of extract/fractions of Q. salicina Blume against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim} 25\;{\mu}g$/mL). Especially, ${\tau}_{50}$ of aglycone fraction in $10 \;{\mu}g$/mL concentration showed the most protective effect at 259.9 min. The inhibitory effects ($IC_{50}$) on tyrosinase and elastase of Q. salicina Blume extracts were higher at aglycone fraction (respectively, $21.82 \;{\mu}g$/mL, $41.18\;{\mu}g$/mL). Active component analysis by TLC and HPLC showed quercetin, keampferol, catechin, quercitrin, isoquercitrin, hyperoside, and etc. These results indicate that Q. salicina Blume extract has strong antioxidative activity and can be used as antioxidant. Particularly, aglycone fraction of Q. salicina Blume showed superior antioxdative activity and high inhibitory effect on tyrosinase and elastase. Therefore, aglycone fraction of Q. salicina Blume could be applicable to new functional cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.1
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• pp.25-35
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• 2008

In this study, the antioxidative effects and inhibitory effects on elastase and tyrosinase of Geranium nepalense extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of Geranium nepalense were in the order: 50% ethanol extract(15.0 ${\mu}g/mL$)${\mu}g/mL$). ${\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities($OSC_{50}$) of some Geranium nepalense extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescense assay. The order of ROS scavenging activities were 50% ethanol extract($OSC_{50},\;0.23{\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Deglycosylated flavonoid fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Geranium nepalense on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Geranium nepalense extracts suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated flavonoid fraction exhibited the most prominent celluar protective effect (${\tau}_{50}$, 676.7 min at 50 ${\mu}g/mL$). The inhibitory effect of aglycone fraction on tyrosinase($IC_{50}$, 70.0 ${\mu}g/mL$) and elastase ($IC_{50}$, 19.9 ${\mu}g/mL$) was very high. Aglycone fractions obtained from the deglycosylation reaction of ethyl-acetate fraction among the Geranium nepalense extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (370 nm). Two components were identified as quercetin(composition ratio, 15.3%), kaempferol(82.8%). These results indicate that extract/fractions of Geranium nepalense 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. And component analysis of Geranium nepalense extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

• Journal of Life Science
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• v.24 no.9
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• pp.935-945
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• 2014

Persimmon leaves were commonly consumed as beverages, but were also used as popular folk medicine in Asia. The purpose of this work was to assess the biological activities of Diospyros Lotus L. extracts (DLLE). Various solvent extracts, including n-Hexnae, $CHCl_3$, EtOAc, and n-BuOH fractions, were obtained from the methanol extract of Diospyros Lotus L. leaves. The increasing interest in the powerful biological activity of plant phenolics and flavonoids outlined the necessity for determining their content in medicinal herbs. In this study, the total polyphenol and flavonoid contents (TPC and TFC) in the EA fraction were higher than those of other fractions. The biological activities of DLLE were tested using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay, as well as superoxide dismutase (SOD) activity as an anti-oxidant effect and ${\alpha}$-glucosidase inhibitory activity as an anti-diabetic effect. The EA fraction with high TPC and TFC values showed the highest anti-oxidant effect and high ${\alpha}$-glucosidase inhibition. The EA fractions were further purified into eight fractions using open column chromatography. Higher anti-oxidant and anti-${\alpha}$-glucosidase activity were observed in polar fractions. The content of the flavonoids, including quercein-3-O-rutinoside, kaempferol-3-O-glucoside, myricetin, luteolin, and kaempferol, were analyzed in effective fractions using high-performance liquid chromatography (HPLC). The results suggest that DLLE have anti-oxidative and anti-diabetic effects and thus, have the potential as anti-diabetic materials and as a source for natural health products.

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.710-718
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• 2002

It is reported that Pueraria Radix contains phtoestrogens whereas flower, and bud of Pueraria lobata Ohwi were not known. In the present study, we determined the amount of phytoestrogen in each portion of P. lobata Ohwi and carried out therapeutic effects of osteoporosis. The amounts of genistein, daidzein, and formononetin in Pueraria Radix (PR), Pueraria Flos (PF), and young Pueratia Folium (PL) were quantitated using a HPLC system. Proliferation of osteoblast and growth inhibitory effect on osteoclast were measured in order to screen their effects on osteoporosis. Proliferation of osteoblast-like cells (Saos-2) was analyzed by both MTT methods and alkaline phosphatase (ALP) assays. Growth inhibitory effect on osteoclast was also detected as Tartrate resistant acid phosphatase (TRAP) assay. Ovariectomized rat as an in vivo animal model was selected and administrations of PR were 1 g/kg/day (PR-1) and 5 g/kg/day (PR-5) for 9 weeks, respectively. Trabecular bone areas (TBAs) of tibia and lumbar were analyzed usibg histomorphological methods. Results show that PR contains the highest level of daidzein ($10435{\pm}2143\;mg/kg$ of dried herb) and stimulated ALP activity, approximately 160% of the control. Growth inhibitory effect on osteoclast by both PR and daidzein were almost identical with control although $IC_{50}$ of genistein was $5.81{\times}10^{-7}$ M. Increases in body weight of OVX rats were suppressed by administration of PR but wet weights of uterus in PR-5 group were increased (p<0.05). Plasma ALP and HDL-cholesterol levels were decreased following ages (p<0.01), and LDL-cholesterol level was also decreased in PR-5 group at 20 week of age (p<0.01). TBAs of tibia and lumbar in PR-1 and PR-5 groups were higher than those of the control although the values were less than those of the sham group (each p<0.01) In conclusion, administrations of PR prevented loss of TBAs of tibia and lumber in OVX rats, while PL and PF did not (p<0.01).

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.6
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• pp.823-831
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• 2015

Evaluation of biological activity and analysis of functional constituents from water and ethanol extracts of four different parts of mulberry (Morus alba L.) tree were carried out to develop functional ingredients and foods using extracts of mulberry tree. The water and ethanol extracts of four different parts of mulberry tree were prepared and their biological activities and functional constituents determined by in vitro assays and HPLC, respectively. In general, ethanol extracts showed stronger biological activities and higher functional constituents than water extracts. Ethanol extracts of mulberry fruit, root bark, and twig showed stronger antioxidant ($IC_{50}=128.4{\mu}g/mL$), ${\alpha}$-glucosidase ($IC_{50}=12.0{\mu}g/mL$), and lipoxygenase ($IC_{50}=36.3{\mu}g/mL$) and tyrosinase ($IC_{50}=410.3{\mu}g/mL$) inhibitory activities, respectively, than those of other parts. Mulberry fruit and leaf showed the highest contents of anthocyanin (cyanidin 3-glucoside: 213.20 mg/100 g) and chlorogenic acid (514.97 mg/100 g), and especially ethanol extract of mulberry leaf contained higher quercetin 3-O-(6-O-malonyl)glucoside (143.25 mg/100 g) and kaempferol 3-O-(6-O-malonyl)glucoside (30.25 mg/100 g) contents without water extract of mulberry leaf. Meanwhile, mulberry twig contained both oxyresveratrol glycoside (48.90 mg/100 g) and its aglycone (21.88 mg/100 g), whereas mulberry root bark contained mostly oxyresveratrol glycoside (724.05 mg/100 g). Additionally, mulberry root bark and leaf contained much higher ${\gamma}$-aminobutyric acid (223.90 mg/100 g) and 1-deoxynojirimycin (86.07 mg/100 g) contents, respectively, than other parts of mulberry tree. These results suggest that high quality processed foods and functional foods using mixtures of mulberry fruits, leaves, twigs, and root barks should be developed for prevention and inhibition of several pathological disorders.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.3
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• pp.189-200
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• 2008

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Quercus glauca extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ of extract I fractions of Quercus glauca leaf was in the order: 50% ethanol extract $(12.45{\mu}g/mL)$ < ethyl acetate fraction $(10.47{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(8.57{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Quercus glauca leaf 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 50% ethanol extract $(OSC_{50},\;4.2{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(1.58{\mu}ug/mL)$ < ethyl acetate fraction $(0.66{\mu}g/mL)$. Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract / fractions of Quercus glauca leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Quercus glauca leaf extracts suppressed photohemolysis in a dose dependent manner, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect $({\tau}_{50}$, 398.67 min at $50{\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Quercus glauca leaf extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (360 nm) as well. Two components were identified as quercetin (55.77%), and kaempferol (44.23 %). TLC chromatogram of ethyl acetate fraction of Quercus glauca leaf extracts revealed 6 bands $(QG1{\sim}QG6)$, Among them, isoquercitrin (QG3), hyperin (QG4), and rutin (QG6) were identified. The inhibitory effect of aglycone fraction on tyrosinase $(IC_{50},\;73.5{\mu}g/mL)$ and elastase $(IC_{50},\;16.2{\mu}g/mL)$ was high. These results indicate that extract / fractions of Quercus glauca 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. And component analysis of Quercus glauca leaf extract and inhibitory activity on tyeisinase and elastase of the aglycone fraction could be applicable to new functional cosmetics.