• Title/Summary/Keyword: 컬럼

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Inhibitory Effect of Lonicera japonica Extract on MMP-1 Production in Human Dermal Fibroblast (인체 섬유아세포에서 인동덩굴 추출물의 MMP-1 생성 억제효과)

  • Jeong, Yeon Su;Lee, Jin Young;Ko, Jaeyoung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.1
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    • pp.67-72
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    • 2020
  • In this study, we attempted to obtain an active ingredient that inhibits the production of matrix metalloproteinase-1 (MMP-1) that breaks down collagen in human skin fibroblasts. More than 50 plant extracts were screened, and Lonicera japonica was selected for this study. The stem of L. japonica was extracted with 70% ethanol and fractions with solvents in the order of hexane, ethyl acetate, and butanol. MMP-1 production were significantly inhibited at the concentration of 50 ㎍/mL of the ethyl acetate layer and 200 ㎍/mL of the butanol layer. To get a fraction containing all of these effective components, 80% ethanol fraction (LJ F80) was obtained through HP20 resin column chromatography. The reference substance, loganin and LJ F80 inhibited dose-dependently MMP-1 production. At the same concentration, LJ F80 showed a higher inhibitory effect than loganin. The stability of this fraction was analyzed with HPLC while kept storing at 4 ℃, room temperature, and 40 ℃, for 16 week. The stability was maintained as ± 10% of initial value with reference loganin. Therefore, it is thought that LJ F80 of L. japonica may be used to improve wrinkles.

Antioxidant activity of partially characterized polysaccharides from the edible mushroom Pleurotus djamor var. roseus

  • Raman, Jegadeesh;Sivakumar, Archana;Lakshmanan, Hariprasath;Raaman, Nanjian;Shin, Hyun-Jae
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.140-149
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    • 2021
  • Mushroom-derived polysaccharides, which are the primary bioactive constituents, are beneficial for human health. Polysaccharides have immuno-modulation, antitumor, and antioxidant properties. Additionally, they have antiviral properties and protect against chronic radiation stress. In this study, high yield water-soluble polysaccharides were obtained from Pleurotus djamor var. roseus basidiocarps. The crude polysaccharide (CP) was extracted sequentially by hot water and ethanol precipitation. The yield of the brown CPs was 5.6% dw. Diethylaminoethyl cellulose and Sepharose-6B column chromatography of CPs generated several fractions. Total glucan content was determined in all the fractions. The F1 fraction displayed the highest sugar content and was considered as a purified polysaccharide (PP). The total glucan and β-glucan content in the four fractions ranged between 76.85-2.95% and 75.08-1.46%, respectively. The yield of the PPs was 300 mg, and it was obtained as a white powder. The PPs were characterized by Fourier-transform infrared spectroscopy (FTIR) and thin-layer chromatography. The FTIR spectral details confirmed the presence of a xylopentose polysaccharide. The antioxidant activity of the PPs was evaluated using in vitro 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay and superoxide radical scavenging assay. The PPs showed strong DPPH free radical and superoxide anion radical scavenging activities in a dose-dependent manner. Purified PPs free of phenolics, protein, and carbohydrates were mainly responsible for the radical scavenging activity. The data suggest the potential of PPs as natural antioxidants.

A Study on the Anti-Pollution Effect of Kombucha Fraction through AhR Expression (AhR 발현을 통한 콤부차 분획물의 안티폴루션 효과 연구)

  • Kang, Eun-Bin;Hyun, Jin-A;Kwon, Hyun-Ji;Beom, Seok-Hyun;Han, Dong-Geun;Park, Ha-Eun;Kim, Hyun-Jeong;Kwak, Ki-Sung;An, Bong-Jeon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.3
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    • pp.213-218
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    • 2021
  • In this study, Kombucha, a fermented drink by adding beneficial bacteria to green tea or black tea with sugar, was classified using column chromatography, and the presence or absence of quercetin glycoside, a type of flavonoids, was confirmed through TLC. In addition, the anti-pollution effect of protecting and improving the skin from pollution was confirmed using the fraction. Keratinocytes were treated with K-QG to confirm the cell viability, showing a survival rate of 90% or more up to a concentration of 100 ㎍/mL, and the cell viability by benzo[e]pyrene and fine dust stimulation to see the anti-pollution effect was measured. At 100 ㎍/mL, the improvement rates were about 68.79% and 50.68%, respectively. In addition, as a result of confirming the expression of AhR activated by benzo[a]pyrene through Western blot, it showed an inhibition rate of about 31.08% at 100 ㎍/mL compared to the control group. Through the results of this study, K-QG is thought to be valuable as an anti-pollution functional material, protecting and improving skin irritated from benzopyrene and fine dust.

An analysis of benzoic acid, methyl paraben and butyl paraben in soy sauce using isotope dilution liquid chromatography mass spectrometry (동위원소희석 질량분석법을 이용한 간장 중의 안식향산, 메틸파라벤, 부틸파라벤의 분석)

  • Ahn, Seonghee
    • Analytical Science and Technology
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    • v.31 no.6
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    • pp.225-231
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    • 2018
  • Benzoic acid, methyl paraben, and butyl paraben are preservatives that have been used in pharmaceutical, cosmetic, and food products. However, as their toxicities for human have been reported, many nations and organizations including Korea have established a regulation limit for thier usage of these preservatives in food products. The present study developed the isotope dilution liquid chromatography mass spectrometry method for accurate determination of three target preseratives in soysauce. In this study, the isotope dilution liquid chromatography mass spectrometry method was developed for accurate determination of three target preservatives in soy sauce. LC separation was optimized considering the pKa of benzoic acid which is lower than those of methyl and butyl parabens. A C18 column was used with 5 mM ammonium acetate and methanol as mobile phases. Mass spectrometry was operated in negative mode and selected reaction monitoring mode (SRM). Soy sauce sample was cleaned-up with C18 SPE cartridge for removing matrix inferences and color material. Optimized conditions and the method were validated with soy sauce reference materials for the analysis of food preservatives from Health Science Authority in Singapore. The measured values of benzoic acid, methyl and butyl paraben agreed well with reference values within their uncertainties.

Study on Determination of Design Factor of Bioreactor for Sulfate Reduction in Mine Drainage (광산배수 내 황산염 저감을 위한 생물반응기의 설계인자 도출 연구)

  • Kim, Kang-Ho;Kang, Chan-Ung;Kim, Sun-Joon;Kim, Tae-Heok;Ji, Won-Hyun;Jang, Hang-Seok;Park, Hyun-Sung
    • Journal of the Korean Society of Mineral and Energy Resources Engineers
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    • v.55 no.6
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    • pp.527-537
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    • 2018
  • Column tests of a sulfate reducing bacteria (SRB) bioreactor were conducted to determine the design factors for sulfate-rich mine drainage. Various substrates were applied to the bioreactor, including cow manure and its mixture with a mushroom compost, with rice straw and limestone as subsidiary materials. This procedure provided a removal efficiency of up to 82% of the total sulfur with the mixture of cow manure (70%), mushroom compost (10%) and rice straw (20%), and higher efficiencies were observed after 2 days of retention time. In the downflow condition of the flow direction, oxygen supply and re-oxidation of the sulfates occurred, causing a decrease in sulfate removal efficiency. The addition of an inorganic sludge containing heavy metals, which was intended for production of metal-sulfides in the bioreactor, had a negative effect on the long-term operation owing to arsenic release and toxicity to the SRB. The results thus show that a bioreactor using a mixed substrate with cow manure and operating in the downflow direction could reduce sulfates and total dissolved sulfur content; this process confirms the applicability of the SRB bioreactor to sulfate-rich saline drainage.

Anti-inflammatory and Anti-bacterial Constituents from the Extracts of Daucus carota var. sativa Aerial Parts (당근 지상부 추출물 유래 항염 및 항균 활성 성분)

  • Kim, Jung Eun;Jo, Yeon Jeong;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.4
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    • pp.427-436
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    • 2018
  • In this study, we investigated anti-inflammatory and anti-bacterial constituents from Daucus carota var. sativa (carrot) areal parts. For the extract and solvent fractions, the anti-inflammatory activities were examined by measuring the nitric oxide (NO) production using LPS-stimulated RAW 264.7 cells. Among them, the ethyl acetate (EtOAc) fraction decreased the NO level in a dose-dependent manner. To elucidate further anti-inflammatory mechanisms, EtOAc fraction was evaluated by estimating their effects on the production of prostaglandin $E_2$ and pro-inflammatory cytokines as well as on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). As a result, the EtOAc fraction was determined to inhibit the production of $PGE_2$, IL-$1{\beta}$, IL-6 and reduce the iNOS, COX-2 protein expression. Upon the anti-bacterial tests using Staphylococcus epidermidis and Propionibacterium acnes, n-hexane (Hex) and EtOAc fractions showed the most potent activities. Three phytochemicals were isolated form the EtOAc fraction; diosmetin (1), diosmin (2), cynaroside (3). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including $^1H$ and $^{13}C$ NMR spectra, as well as comparison of the data to the literatures. Anti-inflammatory and anti-bacterial effects were studied for the isolates. All of the compounds (1 - 3) decreased the NO production, effectively. Also, compound 3 showed anti-bacterial activity on P. acnes. Based on these results, D. carota var. sativa extract could be potentially applicable as anti-inflammatory and anti-bacterial ingredients in cosmetic formulations.

Roles of sugar chains in immunostimulatory activity of the polysaccharide isolated from Angelica gigas (참당귀에서 분리한 다당의 면역활성에 대한 당쇄의 역할)

  • Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.336-342
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    • 2019
  • To elucidate structure-function relationship of polysaccharide from Angelica gigas, the AGE-2c-I was purified by two successive chromatography steps. AGE-2c-I showed a potent anti-complementary activity in a dose-dependent manner. AGE-2c-I with a molecular weight of 140 kDa comprised four monosaccharides and 13 glycosyl linkages, and strongly reacted with ${\beta}$-glucosyl Yariv reagent. For the fine structure analysis of AGE-2c-I, it was sequentially digested by exo-arabinofuranosidase and endo-galactanase. The results indicated that AGE-2c-I was a typical RG-I polysaccharide with side chains such as highly branched ${\alpha}$-arabinan, ${\beta}$-($1{\rightarrow}4$)-galactan and arabino-${\beta}$-3,6-galactan. To characterize the active moiety of AGE-2c-I, the anti-complementary activities of AGE-2c-I and its subfractions were assayed. It was observed that the anti-complementary activity of AGE-2c-I was due to the entire structure that resembled RG-I. In addition, arabino-${\beta}$-3,6-galactan side chain (GN-I) in AGE-2c-I probably plays a crucial role in the anti-complementary activity, whereas ${\alpha}$-arabinan side chain (AFN-I) consisting of 5-linked Araf and 3,5-branched Araf partially contributes to the activity.

Analytical Method Development of (-)-Epicatechin gallate in Penthorum chinense Pursh Extract using HPLC (HPLC를 이용한 낙지다리 추출물의 (-)-­Epicatechin gallate 분석법 개발)

  • Kwon, Jin Gwan;Jung, Yeon Woo;Seo, Changon;Hong, Seong Su;Choi, Chun Whan;Lee, Ji Eun;Shin, Hyun Tak;Jung, Su Young;Kim, Jin Kyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.1
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    • pp.87-93
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    • 2019
  • This study attempted to eatablish a High Performance Liquid Chromatography (HPLC) analysis method for the determination of (-)-epicatechin gallate as a part of the quality control for the development of functional cosmetic materials from Penthorum chinense Pursh. HPLC was performed on a Unison $US-C_{18}$ column ($4.6{\times}250mm$, $5{\mu}m$) with a gradient elution of 0.05% (v/v) trifluoroacetic acid (TFA) and methyl alcohol at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analyte was detected at 280 nm. The HPLC method was performed in accordance with the International Conference on Harmonization (ICH) guideline (version 4, 2005) of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation were 0.11 and 0.33 mg/mL, respectively. Calibration curves showed good linearity ($r^2$ > 0.9999), and the precision of analysis was satisfied (less than 0.6%). Recoveries of quantified compounds ranged from 99.51 to 101.92%. This result indicates that the established HPLC method is very useful for the determination of marker compound in P. chinense Pursh extracts.

Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3 (해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성)

  • Kim, Da Som;Kim, Jong-Hee;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.329-336
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    • 2021
  • Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 ℃. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

Analysis of Cadaverine and Its Worker Honeybee Venom Content (Apis mellifera L.) (꿀벌(Apis mellifera L.) 일벌독의 생체아민 cadaverine 함량 및 분석법)

  • Choi, Hong Min;Kim, Hyo Young;Kim, Se Gun;Han, Sang Mi
    • Journal of Life Science
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    • v.31 no.2
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    • pp.144-148
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    • 2021
  • This study aimed to analyze the content and composition of a biological amine, cadaverine, isolated from the venom of worker honeybees (Apis mellifera L.). This biological amine―which has diverse functionality, such as anti-inflammatory and antibacterial effects―has not been previously reported in bee venom. An assay completed in 13 minutes was developed for the cadaverine present in the bee venom using an ultra-performance liquid chromatograph and a Halo C18 column with acetonitrile and water as the mobile phase. The specificity, accuracy, and precision of the assay were verified, and the assay was validated. The linearity for cadaverine in the bee venom was R2=0.99 or above, indicating a moderate level. The limit of detection and limit of quantification were both 0.3 ㎍/ml, and the rate of recovery was 97.6%-99.1%. The relative standard deviation (RSD) of the intra-day precision and inter-day precision for cadaverine was 0.25%-0.44% and 0.25%-1.25%, respectively, with an RSD that fell within 5% indicating excellent precision. Through this novel assay, it was found that the mean content of cadaverine was 1.10±0.05 mg/g. Our results indicated that the linearity, limit of detection, limit of quantification, and rate of recovery of the cadaverine assay were of a satisfactory level, and the cadaverine content of the bee venom was ably determined. This study provides basic data on cadaverine in bee venom, which will prove useful in further studies on the bioactivity of this component.