• Title/Summary/Keyword: LC-NMR

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Isolation and Purification of Garlic Specific Organic Compounds (마늘 특이 유기화합물의 분리 및 정제)

  • Oh, Tae-Young;Kyung, Kyu-Hang
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.553-557
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    • 2011
  • Garlic specific organic compounds were separated and purified using a recycling preparative high-performance liquid chromatography (HPLC) from blanched garlic cloves. Identification of the compounds involved comparing the previously reported HPLC retention times as well as other identification methods including $^1H$- and $^{13}C$-nuclear magnetic resonance and liquid chromatography-mass spectrometry. The yields of garlic specific organic compounds were 12.2, 42.5, 1.6, 1.2, and 4.8% on wet weight basis of garlic for alliin(S-allyl-L-cysteine sulfoxide), isoalliin(S-1-propenyl-L-cysteine sulfoxide), ${\gamma}$-glutamyl-S-allylcysteine, ${\gamma}$-glutamyl-S-1-propenylcysteine and ${\gamma}$-glutamyl-phenylalanine, respectively. All the compounds, except for ${\gamma}$-glutamylphenylalanine, contained sulfur.

Formation of Flavone Di-O-Glucosides Using a Glycosyltransferase from Bacillus cereus

  • Ahn, Byoung-Chan;Kim, Bong-Gyu;Jeon, Young-Min;Lee, Eun-Jeong;Lim, Yoong-Ho;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.387-390
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    • 2009
  • Microbial UDP-glycosyltransferases can convert many small lipophilic compounds into glycons using uridine-diphosphate-activated sugars. The glycosylation of flavonoids affects solubility, stability, and bioavailability. The gene encoding the UDP-glycosyltransferase from Bacillus cereus, BcGT-3, was cloned by PCR and sequenced. BcGT-3 was expressed in Escherichia coli BL21(DE3) with a glutathione S-transferase tag and purified using a glutathione S-transferase affinity column. BcGT-3 was tested for activity on several substrates including genistein, kaempferol, luteolin, naringenin, and quercetin. Flavonols were the best substrates for BcGT-3. The enzyme dominantly glycosylated the 3-hydroxyl group, but the 7-hydroxyl group was glycosylated when the 3-hydroxyl group was not available. The kaempferol reaction products were identified as kaempferol-3-O-glucoside and kaempferol-3,7-O-diglucoside. Kaempferol was the most effective substrate tested. Based on HPLC, LC/MS, and NMR analyses of the reaction products, we conclude that BcGT-3 can be used for the synthesis of kaempferol 3,7-O-diglucose.

Production and Purification of tazane Derivatives from the Plane Cell Cultures of Taxus Chinensis in Large-scale Process (식물세포 Taxus chinensis 의 대량 배양액으로부터의 Taxane 유도체 생산 및 정체)

  • 김진현
    • KSBB Journal
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    • v.15 no.4
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    • pp.398-401
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    • 2000
  • Taxiods inclusive paclitaxel were produced isolated and purified from plant cell cultures of Taxus chinensis in large-scale process. their structures were elucidated by spectroscopic analyses. These compounds were exactly identical as those in previous studies from the other biomasses of Taxus chinensis and also other species. Also the concentrations of these compounds were compared with the concentration of the paclitaxel in various batches of plant cell cultures. As paclitaxel concentration increased at the end of cell cultures. the concentrations of the other paclitaxel derivatives decreased. The profile of these taxoids production can provide information for better understanding of structure-activity relationships and biosynthesis Importantly it can be utilized as an useful parameter for the quality control of paclitaxel production.

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Biosynthesis of Bile Acids in a Variety of Marine Bacterial Taxa

  • Kim, Doc-Kyu;Lee, Jong-Suk;Kim, Ji-Young;Kang, So-Jung;Yoon, Jung-Hoon;Kim, Won-Gon;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.403-407
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    • 2007
  • Several marine. bacterial strains, which were isolated from seawater off the island Dokdo, Korea, were screened to find new bioactive compounds such as antibiotics. Among them, Donghaeana dokdonensis strain DSW-6 was found to produce antibacterial agents, and the agents were then purified and analyzed by LC-MS/MS and 1D- and 2D-NMR spectrometries. The bioactive compounds were successfully identified as cholic acid and glycine-conjugated glycocholic acid, the $7{\alpha}$-dehydroxylated derivatives (deoxycholic acid and glycodeoxycholic acid) of which were also detected in relatively small amounts. Other marine isolates, taxonomically different from DSW-6, were also able to produce the compounds in a quite different production ratio from DSW-6. As far as we are aware of, these bile acids are produced by specific members of the genus Streptomyces and Myroides, and thought to be general secondary metabolites produced by a variety of bacterial taxa that are widely distributed in the sea.

Mithramycin Inhibits Etoposide Resistance in Glucose-deprived HT-29 Human Colon Carcinoma Cells

  • Lee, Eun-Mi;Park, Hae-Ryong;Hwang, Ji-Hwan;Park, Dong-Jin;Chang, Kyu-Seob;Kim, Chang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1856-1861
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    • 2007
  • Physiological cell conditions such as glucose deprivation and hypoxia play roles in the development of drug resistance in solid tumors. These tumor-specific conditions cause decreased expression of DNA topoisomerase $II{\alpha}$, rendering cells resistant to topo II target drugs such as etoposide. Thus, targeting tumor-specific conditions such as a low glucose environment may be a novel strategy in the development of anticancer drugs. On this basis, we established a novel screening program for anticancer agents with preferential cytotoxic activity in cancer cells under glucose-deprived conditions. We recently isolated an active compound, AA-98, from Streptomyces sp. AA030098 that can prevent stress-induced etoposide resistance in vitro. Furthermore, LC-MS and various NMR spectroscopic methods identified AA-98 as mithramycin, which belongs to the aureolic acid group of antitumor compounds. We found that mithramycin prevents the etoposide resistance that is induced by glucose deprivation. The etoposide-chemosensitive action of mithramycin was just dependent on strict low glucose conditions, and resulted in the selective cell death of etoposide-resistant HT-29 human colon cancer cells.

Metabolism and Excretion Study of DW116, A New Fluoroquinolone, in Rats

  • Jung, Byung-Hwa;Park, Young-Han;Park, Jongsei;Chung, Bong-Chul
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.358-362
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    • 1997
  • Metabolite identification and urinary and biliary excretion of the new fluoroquinolone antibacterial agent DW116 [1-(5-fluoro-2-pyridyl)-6-fluoro-7-(4-methyl-1 -piperazinyl)-1, 4-dihydro-4-oxoquinoline-3-carboxylic acid, hydrochloride] after oral administration have been studied in Sprague-Dawley rats. The excretion kinetics were monoexponential. Most of the drug was eliminated via the hepatic and renal routes. Mean renal clearance of DW116 was 73.4 ml/hr/kg and mean biliary clearance was 83.8 ml/hr/kg. The major metabolite excreted in the bile was identified as the glucuronide ester of the parent drug using base-hydrolysis of the conjugate metabolite followed by co-HPLC with standard compound, $^{19}$ F-NMR and LC-MS methods. The glucuronide conjugate was also found in urine. The mean urinary recoveries of free and total (free plus glucuronide ester) DW116 were $28.6{\pm}2.7% $and $36.4{\pm}1.8%$ of the administered dose and the corresponding biliary recoveries were $14.4{\pm} 5.5%$ and $37.0{\pm}7.6%$, respectively.

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Chemical Constituents of Silene seulensis Nakai from Demilitarized Zone(DMZ)

  • Jung, Yeon Woo;Seo, Chan Gon;Lee, Ji Eun;Hong, Seong Su;Kwon, Jin Gwan;Shin, Hyun Tak;Jung, Su Young;Choi, Jeong Jun;Choi, Chun Whan;Kim, Jin Kyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.92-92
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    • 2018
  • Silene seulensis Nakai was used as traditional medicines in Korea, we collected plant from demilitarized zone (DMZ). S. seulensis was extracted with 30, 50, and 70% ethanol and partitioned successively with n-hexane, EtOAc, dichloromethane and BuOH. These extracts (30, 50 and 70% ethanol) were evaluated the cytotoxicity on B16F10 and Hacat cell lines. The LC-MS/MS data of each fractions (n-hexane, EtOAc, dichloromethane, and BuOH) were compared with MS library, combined with ultraviolet/visual (UV/Vis) and MS data for faster determine structure by database search results. This led to the identification of four compounds (1-4) from S. seulensis. These compounds was isolated first time from S. seulensis. Their chemical structures are elucidated by combinations of NMR and mass spectrometry techniques.

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2'-Hydroxylation of Genistein Enhanced Antioxidant and Antiproliferative Activities in MCF-7 Human Breast Cancer Cells

  • Choi, Jung-Nam;Kim, Doc-Kyu;Choi, Hyung-Kyoon;Yoo, Kyung-Mi;Kim, Ji-Young;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1348-1354
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    • 2009
  • Bioconversion of the isoflavonoid genistein to 2'-hydroxygenistein (2'-HG) was performed using isoflavone 2'-hydroxylase (CYP81E1) heterologously expressed in yeast. A monohydroxylated product was analyzed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and NMR spectrometry and was identified as 2'-HG. An initial bioconversion rate of 6% was increased up to 14% under optimized conditions. After recovery, the biological activity of 2'-HG was evaluated. Bioconverted 2'-HG showed higher antioxidant activity against 1,1-diphenyl-2-picryl hydrazine (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals than did genistein. Furthermore, 2'-HG exhibited greater antiproliferative effects in MCF-7 human breast cancer cells than did genistein. These results suggest that 2'-hydroxylation of genistein enhanced its antioxidant activity and cell cytotoxicity in MCF-7 human breast cancer cells.

Application of Metabolomics to Quality Control of Natural Product Derived Medicines

  • Lee, Kyung-Min;Jeon, Jun-Yeong;Lee, Byeong-Ju;Lee, Hwanhui;Choi, Hyung-Kyoon
    • Biomolecules & Therapeutics
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    • v.25 no.6
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    • pp.559-568
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    • 2017
  • Metabolomics has been used as a powerful tool for the analysis and quality assessment of the natural product (NP)-derived medicines. It is increasingly being used in the quality control and standardization of NP-derived medicines because they are composed of hundreds of natural compounds. The most common techniques that are used in metabolomics consist of NMR, GC-MS, and LC-MS in combination with multivariate statistical analyses including principal components analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). Currently, the quality control of the NP-derived medicines is usually conducted using HPLC and is specified by one or two indicators. To create a superior quality control framework and avoid adulterated drugs, it is necessary to be able to determine and establish standards based on multiple ingredients using metabolic profiling and fingerprinting. Therefore, the application of various analytical tools in the quality control of NP-derived medicines forms the major part of this review. $Veregen^{(R)}$ (Medigene AG, Planegg/Martinsried, Germany), which is the first botanical prescription drug approved by US Food and Drug Administration, is reviewed as an example that will hopefully provide future directions and perspectives on metabolomics technologies available for the quality control of NP-derived medicines.

Phytochemical Constituents from the Rhizomes of Osmunda japonica Thunb and Their Anti-oxidant Activity

  • Woo, Kyeong wan;Jung, Ja Kyun;Lee, Hyun Joo;Kim, Tae Muk;Kim, Min Suk;Jung, Ho Kyung;An, Byeongkwan;Ham, Seong Ho;Jeon, Byung Hun;Cho, Hyun Woo
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.217-221
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    • 2017
  • Eleven compounds (1-11) were isolated from the rhizomes of Osmunda japonica, and their structures were elucidated based on $^1H$, $^{13}C-NMR$ and LC-IT-TOF MS data. Of these compounds, all compounds (1 - 11) have been previously reported, although five (6 - 9, 11) have not previously been isolated from this plant. The antioxidant activities of isolated compounds (1 - 11) were measured by DPPH and ABTS assays, and compound 10 showed the high antioxidant activity.