• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Analytical Science and Technology
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• v.32 no.2
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• pp.56-64
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• 2019

Recently, a number of adulterated products, which are advertised as hair-growth enhancer have been emerged among those who suffer hair loss disease. For continuous control of illegal products, in this study, a rapid and sensitive method for simultaneous screening of 12 compounds that enhance hair-growth was established to protect public health by ultrahigh-performance liquid chromatography coupled to quadrupole-orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS). Fragmentation pathways of them were proposed based on $MS^2$ spectral data obtained using the established method. In this analysis, the LODs and LOQs ranged from 0.05 to 50 ng/mL and from 0.17 to 167 ng/mL, respectively. The square of the linear correlation coefficient ($R^2$) was determined as more than 0.995. The intra- and inter-assay accuracies were respective 88-112 % and 88-115 %. Their precision values were measured within 5 % (intra-day) and 10 % (inter-day). Mean recoveries of target compounds in adulterated products ranged from 84 to 115%. The relative standard deviation of stability was less than 12 % at $4^{\circ}C$ for 48 h. The method was employed to screen 14 dietary supplements advertised to be effective for the treatment of hair loss. Some of the products (~21 %) were proven to contain synthetic drugs that promote hair growth such as triaminodil, minoxidil, and finasteride.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.6
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• pp.927-937
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• 2021

This study was performed to screen the antimicrobial activities of the extract from the Pacific oyster Crassostrea gigas against skin pathogens and to purify the relevant antibacterial peptide. The acidified extract showed potent antibacterial activities against gram-positive and gram-negative bacteria but showed no activity against Candida albicans and no significant cell toxicity. Among acne-causing pathogens, the acidified extract showed potent antibacterial activity only against Staphylococcus aureus, and its antibacterial activity was completely abolished by treatment with trypsin or chymotrypsin, and was inhibited by salt treatment. The acidified extract showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. Based on antimicrobial activity screening and cytotoxic effects, a novel antibacterial peptide was purified from the acidified gill extract using solid-phase extraction, cation-exchange, and reversed-phase HPLC. The resulting peptide had a molecular weight of 4800.8 Da and showed partial sequence homology with the carbonic anhydrase 4 (CA4) protein in the hard-shelled mussel. Overall, we purified a novel antibacterial peptide, named cgCAFLP, which is related to carbonic anhydrase 4 (CA4) protein, against skin pathogens. Our results suggest that the Pacific oyster extract could be used as an additive to control some acne-related skin pathogens (S. aureus).

• Journal of Food Hygiene and Safety
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• v.18 no.4
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• pp.195-201
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• 2003

An easily available, simultaneous identification/determination procedure for sildenafil, homosildenafil, tadalafil, vardenafil in adulterated health related foods was established by using a combination of three different analytical methods; thin layer chromatography(TLC), liquied chromatography-mass spectrometry (LC/MS) and high-performance liquied chromatography (HPLC)/photo-diode-array detector. The sample solution for TLC was applied to silica gel 60 $F_{254}$ plates with ethylacetate/acetonitrile/25%ammonia (90:10:5) as a developing solvent. Spots were located under UV radiation at 254 nm and dragendolfs reagent. Mass spectra of the compounds by LC/MS were investigated with electrospray ionization (ESI) interface, under positive ion mode. The HPLC analysis was performed on a column of capcell pack $C_{18}$ (UG120, 4.6${\times}$250mm I.D. 5 ${\mu}$m)with 0.1% sodium 1-hexansulfonate (in 0.1% phosphoric acid)/acetnitrile (73:27) as a mobile phase, and effluent was minitored with a photo-diode-applied to commercial foods, Sildenafil content was inthe range of 0.4mg/g~360.9 mg/g from 7 out of 35 samples. Homosildenafil content was in the range of 2.2 mg/g~336.0 mg/g from 7 out of 35 samples. Tadalafil content was 429.3 mg/g, 9.6 mg/500 mg from 2 out of 35 samples. The procedure described here is available for the screening of sildenafil, homosildenafil, tadalafil, vardenafil.

• Microbiology and Biotechnology Letters
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• v.36 no.1
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• pp.55-60
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• 2008

Colored rices are a hulled grains having red or purple pigments in bran. Especially black rice (Heugjinjubyeo) is considered to be a healthy food in Asia. Black rice is of great interesting because of the possible biological activity with their anthocyanins. Anthocyanins are water-soluble plant pigments and representatives of flavonoids. The anthocyanins in black rice include cyanidin 3-O-glucoside, peonidin 3-O-glucoside, malvidin 3-O-glucoside, pelagonidin 3-O-glucoside and delphinidin 3-O-glucoside. In this study, anthocyanins in a black rice were analyzed quantitatively and qualitatively with HPLC and UV-Vis spectrophotometer. The anthocyanins contained approximately 95% of cyanidin-3-O-glucoside and 5% of peonidin-3-O-glucoside. Antioxidant activities of the anthocyanin extract were investigated by using various in vitro methods. The 100g/ml concentration of the anthocyanin extracted exhibited 88.83% inhibition on the peroxidation of linoleic acid, 55.20% DPPH free radical scavenging activity, 54.96% superoxide anion radical scavenging activity, and 72.67% hydrogen peroxide scavenging activity. And it also showed high ferrous ion reducing capability. These results suggest that the anthocyanin extracted from black rice may be utilized as a possible antioxdiant agent against ROS.

• Journal of Life Science
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• v.21 no.9
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• pp.1244-1250
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• 2011

The study was performed to evaluate the antibacterial and antiviral activities of ginseng fine root in order to search for antibacterial substances. Among 8 kinds of fermentation strains, Lactobacillus plantarum was selected based on viable cell count and antibacterial activities during incubation. Optimum conditions of ginseng fine root fermentation for L. plantarum were incubation at $35^{\circ}C$ for 48 hr in 5% ginseng fine root broth. That methanolic extract of fermented ginseng fine root broth was observed to be antibacterial and have antiviral activities. The results of paper disc method of non-fermented extract and fermented extract measured against E. coli was 11 mm and 20 mm, S. aureus was 15 mm and 22 mm, respectively. Shaking flask method was observed to inhibit the growth E. coli and S. aureus in fermented extract by 99.9%. However, antiviral activity of Feline calicivirus (FCV) was mostly activated. Fermented extract was used to investigate the compositional changes of ginsenosides on HPLC analysis. By fermentation, ginsenoside Rg1, Re and Rd were increased, with Rd showing a significant increase of 50 ${\mu}g/g$. These results suggest that ginseng fine root extract is a useful resource.

• Journal of Environmental Science International
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• v.17 no.1
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• pp.19-27
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• 2008

Artichoke extracts are widely used alone or in association with other herbs for embittering alcoholic and soft drinks and to prepare herbal teas or herbal medicinal products in Viet Nam. The objective of this paper was a screening of flavonoids and total phenolic compounds content in the parts of artichoke (Cynara scolymus L.) as flowers, leaves, roots, trunks, stumps, The total phenolic compounds and flavonoids in the parts of artichoke were extracted among 3 extraction methods as methanol extraction (EM1), mixing methanol and water method (EM2) and water extraction method (EM3). Total phenolic compounds and flavonoids were determined by UV/VIS, HPLC techniques. The apigenin 7-O-glucosides, cynarin, narirutin, gallic acid, caffeic acid were found as the main flavonoids constituents in all parts of artichoke. It showed that value of total phenolic compounds and flavonoids by EM3 were higher than that of total phenolic compounds and flavonoids by EM1 and EM2. Furthermore, the results of this study revealed that total phenolic compounds and flavonoids, obtained by these convenient extraction methods, may show the quick efficacy of artichoke in all respects of their quality and quantity.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.336-336
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• 1994

In order to develope the in vitro short term screen-ing method for carcinogen, we studied a purification method for thymine glycol in oxidaized DNA. Thymine glycol (5,6-dihydroxy-5, 6-dihydrothymine) is the major stable radiolysis poduct in thymine by chemical oxidants and ionzing radiation and it is a useful biomarker among oxidized DNA adducts, related with carcinogenests. Standard thymine glycol was prepared by oxidation of 〔$^3$H〕 thymine with KMnO$_4$ followed by purification with HPLC-LSC system and it was assayed by TLC and gas chromatography-MSD. 〔$^3$H〕 DMA adducts was isolated from E. coli (wild type ) treated with oxidative agents such as benzo(a)pyrene, adriamycin, aflatoxin B$_1$ and KBrO$_3$. These oxidative agents generated free radicals in cells by oxidative metabolism. As a result, thymine glycol was produced in cultured E. coli by four chemicals. This result shows that this methodology should be useful tool in screening oxidative carcinogen.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.641-646
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• 1995

During the screening of inhibitors of melanin biosynthesis from microbial secondary metabolites, a fungal strain MR304 which was capable of producing high level of an inhibitor was selected. Based on taxonomic studies, this fungus could be classified as Trichoderma harzianum. The active compound (MR304-1) was purified from culture broth by Diaion HP-20 column chromatography, ethylacetate extraction, Sephadex LH-20 column chromatographv and HPLC. The inhibitor was identified as 3-(1,5-dihvdroxy-3-isocyanocyclopent-(E)-3-envl)prop-2-enoate by spectroscopic methods of UV, ESIMS, $^{1}$H-NMR, $^{13}$C-NMR, NOE, HMQC and HMBC. MR304-1 showed strong mushroom tyrosinase inhibitory activity with IC$_{50}$ value of 0.25 $\mu $g/ml. It inhibited melanin biosynthesis with 15 mm inhibition zone at 30 $\mu $g/paper disc in Streptomyces bikiniensis, a bacterium used as an indicator organism in this work. It also inhibited melanin biosynthesis in B16 melanoma cells with a niinimum inhibitory concentration of 0.05 $\mu $g/ml.

• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1236-1242
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• 2011

Xanthomonas oryzae pv. oryzae (Xoo) is the most devastating pathogen to Oryza sativa and has been shown to cause bacterial blight. Two bioactive compounds showing antimicrobial activities against Xoo strain KACC 10331 were isolated from a Streptomyces bottropensis strain. The ethyl acetate extract was fractionated on a Sephadex LH-20 column, and then purified by preparative HPLC. The purified compounds were identified as bottromycin A2 and dunaimycin D3S by HR/MS and $^1H$ NMR analyses. The MIC value against Xoo and the lowest concentration still capable of suppressing rice bacterial blight were 2 ${\mu}g$/ml and 16 ${\mu}g$/ml for bottromycin A2, and 64 ${\mu}g$/ml and 0.06 ${\mu}g$/ml for dunaimycin D3S, respectively. These two compounds were shown to exert different bioactivities in vitro and in rice leaf explants.