• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.6
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• pp.875-885
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• 2022

This study was performed to confirm the optimal extraction method for antimicrobial peptides from the Hard-shelled mussel. Extractions were performed with two processes including 1% HAc/boiling and 1% HAc/non-boiling methods and used extracts for the comparison of the antimicrobial activity, protease stability, action mechanism, AU-PAGE (acid-urea PAGE), and HPLC chromatograms. 1% HAc/boiling extract showed potent antibacterial activities both against Gram-positive and negative bacterium but 1% HAc/non-boiling extract showed antibacterial activity only against Gram-positive bacteria. Treatment of 1% HAc/boiling extract with proteases retained almost antibacterial activity against B. subtilis, but abolished significant antibacterial activity against E. coli D31. Only 1% HAc/boiling extract showed two discrete clearing antibacterial zones including slow migrating and rapid migrating zones. Both extracts showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. In comparison of the chromatogram obtained from C₁₈ or cation-exchange HPLC, the eluted peaks from 1% HAc/boiling extract showed high hydrophobic property or absorbance compared to 1% HAc/non-boiling extract, respectively. The concentration of the purified antimicrobial peptide was also higher in 1% HAc/boiling extract than in 1% HAc/non-boiling extract. Our results suggest that the effective extraction condition for antimicrobial peptides from marine invertebrate is boiling process in a weak acetic acid solution (1%).

• Analytical Science and Technology
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• v.21 no.5
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• pp.403-411
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• 2008

Dimethyldibutyltetradecylmalonamide (DMDBTDMA) extractant was used in a solvent extraction process for a radioactive liquid waste treatment. For the study of radiolysis phenomena, DMDBTDMA was synthesized and the degradation compounds (n-methylbutylamine, tetradecane, 1-tetradecanol) in the DMDBTDMA extractant, irradiated with $^{60}Co$ gamma ray, were identified and determined as radiolysis products by a Fourier transform infrared (FT-IR), gas chromatograph/mass spectrometer (GC/MS) analysis and GC/MS with selected ion monitoring (SIM) mode. Retention behavior of n-methylbutylamine, n-dodecane, tetradecane and 1-tetradecanol in the total ion chromatogram with the standard materials and n-dodecane as the internal standard (ISTD) were 2.35 min., 8.83 min., 10.68 min. and 12.75 min., respectively. In the case of tetradecane, there was a linear relationship between the concentration of the tetradecane and the absorbed dose of the ${\gamma}$-ray irradiated DMDBTDMA.

• FOCUS: LIFE SCIENCE
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• no.1
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• pp.1.1-1.4
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• 2024

This article provides comprehensive guidance on the maintenance, cleaning, regeneration, and storage of silica-based HPLC (High-Performance Liquid Chromatography) columns. The general considerations emphasize the importance of using in-line filters and guard cartridges to protect columns from blockage and irreversible sample adsorption. While these measures help, contamination by strongly adsorbed sample components can still occur over time, leading to an increase in back pressure, loss of efficiency, and other issues. To maximize column lifetime, especially with UHPLC (Ultra-High Performance Liquid Chromatography) columns, it is advisable to use ultra-pure solvents, freshly prepared aqueous mobile phases, and to filter all samples, standards, and mobile phases. Additionally, an in-line filter system and sample clean-up on dirty samples are recommended. However, in cases of irreversible compound adsorption or column voiding, regeneration may not be possible. The document also provides specific recommendations for column cleaning procedures, including the flushing procedures for various types of columns such as reversed phase, unbonded silica, bonded normal phase, anion exchange, cation exchange, and size exclusion columns for proteins. The flushing procedures involve using specific solvents in a series to clean and regenerate the columns. It is emphasized that the flow rate during flushing should not exceed the specified limit for the particular column, and the last solvent used should be compatible with the mobile phase. Furthermore, the article outlines the storage conditions for silica based HPLC columns, highlighting the impact of storage conditions on the column's lifetime. It is recommended to flush all buffers, salts, and ion-pairing reagents from the column before storage. The storage solvent should ideally match the one used in the initial column test chromatogram provided by the manufacturer, and column end plugs should be fitted to prevent solvent evaporation and drying out of the packing bed.

• The Korean Journal of Nuclear Medicine
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• v.32 no.5
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• pp.425-432
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• 1998

Purpose: For the purpose of using Re-188 as a therapeutic radionuclide, we performed the quality control of the W-188/Re-188 generator system. Materials and Methods: Several quality control tests of the Re-188 eluate from generator were carried out for about 300 days. After elution of Re-188 with normal saline (20 ml), chromatogram and gamma-ray spectrum of Re-188 eluate were obtained. The presence of aluminum which was derived from the alumina bed of the generator was detected by using aluminum ion indicator kit. Re-188 eluate was allowed to decay for several days, and then W-188 breakthrough in the Re-188 eluate was measured by detecting gamma-ray at 227 keY and 290 keV. The pH and the pyrogenicity of the eluate were checked. The Re-188 bolus was concentrated with ion exchange columns. Results: The radioactivity of Re-188 eluate from the generator was $67.4{\pm}7.0%$ of W-188 during 270 days, and it was highest at third day after previous elution. Radiochemical purity of Re-188 eluate obtained from chromatogram was higher than 99%. Gamma-ray spectrum of Re-188 eluate showed a peak at 155 keV. Aluminum ion and W-188 contamination were not detected. The PH of Re-188 eluate was 3 and the concentration yield was 85%. Conclusion: Our experiments and results on quality control tests of Re-188 eluate from W-188/Re-188 generator may be useful for setting W-188/Re-188 generator in hospitals.

• The Korean Journal of Physiology
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• v.23 no.1
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• pp.79-87
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• 1989

Molecular property as well as biological activities of bombesin-like substance (BBS-LS) isolated from the skin of B. orientalis in Korea was compared with those of synthetic BBS-14. BBS-LS in the crude extract was applied on a Sephadex G-50, superfine column $(1.6{\times}112\;cm)$. On chromatogram, the first peak (3.5% of BBS-LS) was found ahead of synthetic GRP-27, and the second peak (rest) between synthetic GRP-27 and synthetic BBS-14. The main form of BBS-LS was successfully purified by using a column of alkaline alumina followed by sequential gel-filtrations on a column of Sephadex G-10, fine and a column of Sephadex G-50, superfine. Chromatographic analysis of the purified BBS-LS using a column of Sephadex G-50, superfine and reversed phase HPLC revealed that the main form of BBS-LS in the skin of B. orientalis could be distinctly different from either BBS-14 or GRP-27 in molecular size. The purified BBS-LS exerted biological activities quite identical to those of synthetic BBS-14. The results of the present investigation indicate that the skin of B. orientalis contains BBS-LS composed of two distinct forms. The main form of BBS-LS purified in the present study is heterogenous to both synthetic BBS-14 and GRP-27 in molecular size but identical to BBS in biological activities.

• Applied Chemistry for Engineering
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• v.23 no.1
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• pp.93-99
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• 2012

In this study, the evaluation of antioxidative activity and componential analysis of C. obtusa leaf extracts was carried out. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of C. obtusa leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction ($OSC_{50}$; 0.22 ${\mu}g/mL$) and aglycone fraction of C. obtusa leaf extracts (0.20 ${\mu}g/mL$) showed about 7 times more prominent ROS scavenging activity than L-ascorbic acid (1.50 ${\mu}g/mL$). The cellular protective effects of fractions obtained from C. obtusa leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction and aglycone fraction of C. obtusa leaf extracts showed the cellular protective effects in a concentration dependent manner (5~25 ${\mu}g/mL$). The inhibitory effect ($IC_{50}$) of ethyl acetate fraction and aglycone fraction on tyrosinase exhibited 74.43 and 53.80 ${\mu}g/mL$, repectively. The aglycone fraction showed four times higher tyrosinase inhibitory effect than arbutin (226.88 ${\mu}g/mL$), known as a whitening agent. The aglycone fraction of C. obtusa leaf extracts showed three bands in TLC chromatogram and three peaks in HPLC chromatogram (360 nm). Three compounds were identified as taxifolin, quercetin and kaempferol. These results indicate that the fractions of C. obtusa leaf extracts 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 reactive oxygen species. The fractions of C. obtusa leaf extracts can be applicable to new functional cosmetics for antioxidan and whitening effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.1
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• pp.47-55
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• 2009

In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component analysis of fermented Melissa officinalis extracts were investigated. The ethyl acetate fraction of fermented extract ($8.38{\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of M. officinalis. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some M. officinalis extracts on ROS generated in $Fe^{3+}$-EDTA/$H_{2}O_{2}$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract ($0.63{\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of M. officinalis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The M. officinalis extracts suppressed photohemolysis in a concentration dependent manner ($5\;{\sim}\;75{\mu}g/mL$). The inhibitory effect of M. officinalis extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase of some M. officinalis extracts was 50 % ethanol extract ($365{\mu}g/mL$) < ethyl acetate fraction of fermented extract ($122.43{\mu}g/mL$) < ethylacetate fraction ($94.8{\mu}g/mL$). Fractions of ethyl acetate both from ordinary and fermented M. officinalis extracts showed 2 band in TLC and 2 peak in HPLC (330 nm). In HPLC chromatogram of ethyl acetate fraction, peak 1 (51.64 %) and peak 2 (48.36 %) were identified as caffeic acid and rosmarinic acid in the order of elution time. Also, in HPLC chromatogram of ethyl acetate fraction of fermented extract, peak 1 (4.13 %) and peak 2 (95.87 %) were identified as caffeic acid and rosmarinic acid in the order of elution time. These results indicate that the component and content of ordinary and fermented extracts of M. officinalis are different. And the extract of M. officinalis can be used as an antioxidant.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.2
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• pp.117-127
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• 2008

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Cayratia japonica extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities($FSC_{50}$) of extract/fractions of Cayratia japonica were in the order: 50% ethanol extract(114.3 ${\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Reactive oxygen species(ROS) scavenging activities($OSC_{50}$) of some Cayratia japonica extracts in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were deglycosylated flavonoid aglycone fraction($OSC_{50},\;3.30{\mu}g/mL$)<50% ethanol extract(1.21 ${\mu}g/mL$)${\mu}g/mL$). Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Cayratia japonica on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Cayratia japonica extracts suppressed photohemolysis in a concentration dependent manner($1{\sim}25{\mu}g/mL$), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect(${\tau}_{50}$, 175.05min at 25 ${\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Cayratia japonica extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments(360 nm). Two components were identified as luteolin(composition ratio, 47.50%), apigenin(52.50). TLC chromatogram of ethyl acetate fraction of Cayratia japonica extract revealed 3 bands and HPLC chromatogram showed 4 peaks, which were identified as luteolin-7-O-${\beta}$-D-glucopyranoside(composition ratio, 11.14%), apigenin-7-O-${\beta}$-D-glucuronopyranoside(15.38%), luteolin(23.55%) and apigenin(49.92%) in the order of elution time. The inhibitory effect of aglycone fraction on elastase($IC_{50},\;70.5{\mu}g/mL$) was very high. These results indicate that extract/fractions of Cayratia japonica 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 Cayratia japonica extract and antioxidative effects could be applicable to new cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.4
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• pp.251-262
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• 2007

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Aspalathus linearis extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of Aspalathus linearis were in the order: 50 % ethanol extract ($11.50\;{\mu}g/mL$) < deglycosylated flavonoid aglycone fraction ($8.47\;{\mu}g/mL$) < ethylacetate fraction ($4.76\;{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Aspalathus linearis 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 activities were ethylacetate fraction ($OSC_{50},\;4.58\;{\mu}g/mL$) < deglycosylated flavonoid aglycone fraction ($2.20\;{\mu}g/mL$) < 50 % ethanol extract ($1.09\;{\mu}g/mL$). 50 % Ethanol extract showed the most prominent scavenging activity. The protective effects of extract/fractions of Aspalathus linearis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Aspalathus linearis extracts suppressed photohemolysis in a concentration dependent manner, particularly 50 % ethanol extract exhibited the most prominent celluar protective effect (${\tau}_{50}$, 272.00 min at $50\;{\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethylacetate fraction among the Aspalathus linearis extracts, showed 3 bands in TLC and 3 peaks in HPLC experiments (360 nm). Three components were identified as luteolin (composition ratio, 18.24 %), quercetin (58.79), and kaempferol (22.97). TLC chromatogram of ethylacetate fraction of Aspalathus linearis extract revealed 7 bands and HPLC chromatogram showed 9 peaks, which were identified as isoorientin (composition ratio, 14.71 %), orientin (28.84 %), vitexin (5.63 %), rutin and isovitexin (12.73 %), hyperoside (9.24 %), isoquercitrin (5.40 %), luteolin (1.48 %), quercetin (17.61 %) and kaempferol (4.59 %) in the order of elution time. The inhibitory effect of aglycone fraction on elastase ($IC_{50},\;9.08\;{\mu}g/mL$) was very high. These results indicate that extract/fractions of Aspalathus linearis 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 Aspalathus linearis extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.3
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• pp.233-244
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• 2008

In this study, the antioxidative effects, inhibitory effects on elastase and tyrosinase, and component analysis of Psidium guajava leaf extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities $(FSC_{50})$ of extract/fractions of Psidium guajava leaf were in the order: 50% ethanol extract $(7.05{\mu}g/mL)$ < ethyl acetate fraction $(3.36{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(3.24{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Psidium guajava 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 activities were 50% ethanol extract $(OSC_{50},\;2.17{\mu}g/mL)$ < ethyl acetate fraction $(0.64{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(3.39{\mu}g/mL)$. Aglycone fraction showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of Psidium guajava leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Psidium guajava leaf extracts suppressed photohemolysis in a concentration dependent manner $(1{\sim}10{\mu}g/mL)$, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect ${\tau}_{50}\;107.5min\;at\;1{\mu}g/mL)$. Aglycone fraction obtained from the deglycosylation reaction of ethyl acetate fraction among the Psidium guajava leaf extracts, showed 1 band in TLC and 1 peak in HPLC experiments (360 nm). One component was identified as quercetin. TLC chromatogram of ethyl acetate fraction of Psidium guajava leaf extract revealed 5 bands and HPLC chromatogram showed 5 peaks, which were identified as quercetin 3-O-gentobioside (10.32%) , quercetin 3-O-${\beta}$-D-glucoside (isoquercitin, 13.30%), quercetin 3-O-${\beta}$-D-galactoside (hyperin, 11.34%), quercetin 3-O-${\alpha}$-L-arabinoside (guajavarin, 19.70%), quercetin 3-O-${\beta}$-L-rhamnoside (quercitrin, 45.33%) in the order of elution time. The inhibitory effect of Psidium guajava leaf 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. Inhibitory effects $(IC_{50})$ on tyrosinase of some Psidium guajava leaf extracts was 50% ethanol extract $(149.67{\mu}g/mL)$ < ethylacetate fraction $(30.67{\mu}g/mL)$ < deglycosylated aglycone fraction $(17.10{\mu}g/mL)$. Inhibitory effects $(IC_{50})$ on elastase of some Psidium guajava leaf extracts was 50% ethanol extract $(6.60{\mu}g/mL)$ < deglycosylated aglycone fraction $(5.66{\mu}g/mL)$ < ethylacetate fraction $(3.44{\mu}g/mL)$. These results indicate that extract/fractions of Psidium guajava leaf can function as antioxidants in bioloigcal systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Psidium guajava leaf extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.