• Title/Summary/Keyword: HP-TLC

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Improved Method for the Preparation of Crude Ginseng Saponin (인삼 조사포닌의 조제 방법 개선)

  • 김시관;곽이성
    • Journal of Ginseng Research
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    • v.22 no.3
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    • pp.155-160
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    • 1998
  • This stuffy was carried to establish a new efficient method for the preparation of edible crude ginseng saponin. The conventional butanol extraction and resin adsorption methods were compared for the contents of total crude ginseng saponin and major ginsenosides. Seventy- percent methanol extract was applied to Diaion HP-20 column and the resin was washed with Hn and eluted with absolute methanol. The methanol elute was dried in vivo and analyzed for its ginsenosides. Use of ethanol instead of methanol to make edible crude ginseng saponin gave a similar result. Butanol extraction was performed by the conventional method. The final aqueous layer from butanol extraction was passed through Diaion HP-20 column followed by elution with methanol and Diaion HP-20 passed fraction was extracted with butanol to recover remaining components, respectively, in order to determine saponin loss. TLC and HPLC qualitatively and quantitatively monitored Ginsenosides, respectively. Loss of ginsenosides was higher in butanol extraction method than in Diction HP-20 adsorption method. In addition, saponin fractions prepared by Diction HP-20 adsorption method showed higher content of each ginsenoside, showing 8.2% higher purity than that of butanol extracted fraction. From these results, we propose the resin adsorption method as a new efficient measure for the preparation of crude ginseng saponin, which is edible by using spirit instead of methanol.

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Biosynthesis of $17\alpha-hydroxy$, $20\alpha-dihydroprogesterone$ by Ovaries of the Spotted Flounder (Verasper variegatus)

  • Baek Hea-Ja
    • Fisheries and Aquatic Sciences
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    • v.4 no.2
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    • pp.70-74
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    • 2001
  • To examine the production of steroids with potential oocyte maturation-inducing activity in the spotted flounder, Verasper variegatus, we have incubated post-vitellogenic oocytes (0.82­0.95mm in diameters) with radiolabeled pregnenolone and $17\alpha-hydroxyprogesterone$. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The two main metabolites (progestogens) found in both incubations co-migrated with $17\alpha-hydroxy$, $20\alpha-dihydroprogesterone$ $(17\alpha, 20\alpha OHP)$ and $17\alpha-hydroxy,\;$20\beta-dihydroprogesterone$ (17 a20{30HP). Additional chromatography by HPLC and TLC confirmed the presence of radioactive $17\alpha, 20\alpha OHP$ and a large amount of unknown metabolite. The present study did not reveal in vitro formation of $l7\alpha 20\beta OHP$. Although 1$l7\alpha 20\beta OHP$ was found in a small amount, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process in the spotted flounder.

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Isolation of S-627(A) and (B) as Candidates of Anticandiosis Agent (생리활성물질 S-627(A)와 S-627(B)의 분리)

  • Lee, Sang-Han;Lee, Dong-Sun;Seu, Young-Bae;Kim, Jong-Guk;Hong, Soon-Duck
    • Journal of Life Science
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    • v.7 no.3
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    • pp.186-191
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    • 1997
  • In order to select new anticandiosis agent-producing candidates, we used a modified selection method. Two active fractions designated as S-6279A) and (B) were isolated from the fermentation broth of Streptomycetes sp. S627 which was screened by methanol extraction, diaion HP-20 and silica gel column chromatography (chloroform-me-thanol, and benzene-ethyl acetate), and preparative thin layer chromatography.

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An Isolation of Crude Saponin from Red-Ginseng Efflux by Diaion HP-20 Resin Adsorption Method (홍삼유출액으로부터 Diaion HP-20 수지 흡착법에 의한 조사포닌의 분리)

  • 곽이성;경종수;김시관;위재준
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.1-5
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    • 2001
  • This study was carried out to isolate saponin compounds from red-ginseng efflux, which was produced during the industrial processing of red-ginseng from fresh ginseng. We isolated crude saponin from the efflux extract (moisture content 35.0%) by using Diaion HP-20 adsorption method. Non-saponin fraction, which was adsorbed on Diaion HP-20 resin, was removed by eluating with $H_{2}O$ and 25% spirit. Then crude saponin was eluated with 95% spirit, continuously. Saponin in the eluated fractions was confirmed by TLC analysis. Crude saponin isolated from red ginseng efflux extract contained 12.10% of saponin. whereas those of white ginseng and red-ginseng were 3.30 and 3.39%, respectively. Ginsenoside contents showed the highest contents kin crude saponin from red ginseng efflux extract. Expacilly, the ginsenoside-$Rb_{1}$ and Re showed the highest contents in red-ginseng efflux extract when compared with those of white ginseng and red ginseng crude saponins. And the other ginsenosides except ginsenoside-$Rb_{1}$ and -Re also showed the highest contents in red ginseng efflux extract. However, the ratio of PD saponin (Panaxadiol saponin: $Rb_{1}+Rb_{2}$+Rc+Rd) to PT saponin (panaxatriol: $Re+Rg_{1}$) showed almost the same level when compared with those of ginseng saponin fractions. Ratio of PD/PT from red ginseng efflux extract was 1.99. Ratios of PD/PT from white ginseng and red ginseng were 1.85 and 1.84, respectively. Saponin purity, which was calculated by ratio percent of total ginsenoside to curde saponin content, was 45.90%. In case of white ginseng and red ginseng, the purities were 35.50 and 36.00%, respectively. However, by PHLC analysis, we confirmed that crude saponin isolated from red ginsengs. It suggested that crude saponin isolated from red ginseng ellux also would be useful component as ginseng saponins.

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Analysis of the Content of Hesperidin and Essential Oils from the Peels of Various Citrus Species (진피류(陳皮類) 한약재의 Hesperidin과 정유성분 비교)

  • Ham, In-Hye;Jung, Eui-Dong;Lee, Kyung-Jin;Lee, Je-Hyun;Bu, Young-Min;Kim, Ho-Cheol;Choi, Ho-Young
    • The Korea Journal of Herbology
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    • v.23 no.4
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    • pp.159-170
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    • 2008
  • Objectives: This study was carried out to evaluate the quality of the peels of various Citrus species. Method: The contents of hesperidin from fruit peels used as Citri Pericarpium such as C. natsudaidai, C. grandis, C. unshiu, and C. sunki, were analyzed by HP-TLC, HPLC, and essential oils of those were analyzed by GC/MS. Results: HPLC analysis showed that the hesperidin from the peel of C. unshiu and C. reticulata was satisfied the standard of Korean Pharmacopoeia. The essential oil was analyzed by GC/MS. As a result, limonene, furfural, 5-methyl-2-furfural, linalool oxide(cis), linalool oxide(trans), terpinen-4-ol, $(-)-{\alpha}$-terpineol, germacrene D, 4-methyl-2,6-di-tert-butylphenol was detected in all 4 kinds of Citrus species. Conclusions: As a result of chemotaxonomical similarity analysis with essential oils, the peels of C. natsudaidai and C. grandis are closely related, while C. unshiu is distantly related to the others.

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Antioxidative and Cytotoxicity Activities against Human Colon Cancer Cells Exhibited by Edible Crude Saponins from Soybean Cake (대두박 식용사포닌의 항산화 및 대장암세포 성장 억제효과)

  • Park, Kyung-Uk;Kim, Jae-Yong;Seo, Kwon-Il
    • Food Science and Preservation
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    • v.16 no.5
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    • pp.754-758
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    • 2009
  • To develop soybean cake as a functional food material, the anti-oxidative and cytotoxic activities against human colon cancer cells of crude saponins isolated from 70% (v/v) ethanol extracts of cake were investigated. The Diaion HP-20 adsorption method was used for isolation of crude saponins, which were then eluted with 100% ethanol. The non-saponin fraction was removed by elution with $H_2O$ and 20% (v/v) ethanol. The results of thin layer chromatography (TLC) analysis confirmed that crude saponins were present in the 100% ethanol extract of soybean cake. The hydrogen-donating properties of saponins were more than 60% at a concentration of $1,000\;{\mu}g/mL$. malondialdehyde(MDA) production was $1,200\;{\mu}mol\;MDA/g$ in mouse liver homogenate treated with crude saponins at the concentration of $1,000\;{\mu}g/mL$. This value was lower than that of the control, which was $3,700\;{\mu}mol\;MDA/g$. Saponins inhibited the growth of colon cancer cells in a dose- and time-dependent manner. Saponins also resulted in a decrease in the proportion of cells in the G1 phase of the cell cycle, whereas the cell proportion in G2/M phase was increased with $1,000\;{\mu}g/mL$ saponins. Thus, we conclude that saponins may induce G2/M cell cycle arrest.

New Efficient Method for Isolation and Purification of Ginsenosides (Ginsenoside의 새로운 분리.정제 방법)

  • 김세원;황석연
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.284-288
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    • 1998
  • This study was carried out to establish a new efficient method for isolation and purification of ginsenosides. Silica gel column chromatography, having been used for the isolation of ginsenosides, is advantageous to obtain a large amount of ginsenosides. However, it has a disadvantage to isolate ginsenosides to their highest purity. In addition, normal-or reverse-phase HPLC method thus far reported is confined to quantitative analysis. Especially, it has not been possible to isolate racemic 20(S)- and 20(R)-ginsenoside Rg2. In this experiment, isolation and purification of ginsenosides were accomplished by Diaion HP-20 adsorption chromatography, silica gel column chromatography, recrystalization and Prep. HPLC with or without Prep. TLC. From this study, we could establish a new efficient method for isolation and purification of 9 major and/or minor ginsenosides.

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Pancreatic lipase Inhibitory Compound from Apis mellifera venome

  • Kim, Jun-Ran;Kim, Shin-Duk
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.2
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    • pp.57-59
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    • 2008
  • While searching for pancreatic lipase inhibitors, the active compound was found in a methanol extract of Apis mellifera venome. The active compound was isolated by Diaion HP-20 column chromatography, thin layer chromatography and HPLC. The active compound is stable to the extreme pH and heat. There is no loss of activity both in acidic and alkaline solution in the pH range of 2 to 11 by heating for 15 minutes at $90^{\circ}C$. The rf value of the compound was 0.51 at TLC with butanol : methanol: water (4:1:2) solvent system. The molecular weight of the compound was determined to be 293 by EI-MS.

Characterization and Structural Dtercination of an Antifungal Compound Produced by Pseudomonas aeruginosa KGM-100 (Pseudomonas aeruginosa KGM-100이 생산하는 항생물질의 특성 및 구조)

  • Kim, Kyung-Seok;Hong, Su-Hyung;Lee, Eun-Ju;Park, Yong-Bok;Park, Yong-Tae;Ha, Ji-Hong
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.98-103
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    • 1995
  • During the screening of antifungal antibiotics from microbial metabolites, we selected Pseudomonas aeruginosa KGM-100 showing powerful antagonistic activity against various phytopathogenic fungi. Antibiotics KGM-100A and KGM-100B were purified from the culture broth of Pseudomonas aeruginosa KGM-100 by diaion HP-20 column chromatography, ethyl acetate extraction, silica gel column chromatography, preparative TLC and recrystallization. KGM-100A which was recrystallized in MeOH showed antimicrobial activities against a broad spectrum of fungi and bacteria. Physico-chemical properties of KGM-100A were determined and identified to be phenazine-l-carboxylic acid by UV, IR, $^{1}$H-NMR, $^{13}$C-NMR, mass spectrum, and elemental analyses.

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Purification and Characterization of an Antifungal Antibiotic from Bacillus subtilis LAM 97-44 (Bacillus subtilis LAM 97-44가 생산하는 항진균성 항생물질의 정제 및 특성)

  • Lee, No-Woon;Kwon, Tae-Jong;Yi, Dong-Heui
    • Applied Biological Chemistry
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    • v.46 no.2
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    • pp.69-73
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    • 2003
  • A novel antifungal antibiotic for azole-resistant Candida albicans was purified from the culture broth of Bacillus subtilis LAM 97-44 by butanol extraction, Diaion HP-20 and Dowex-50 adsorption chromatography, silica gel flash chromatography followed by HPLC and designated LAM-44A. LAM-44A was stable for 60 min at $100^{\circ}C$, and pH range from 2 to 10. MIC values were observed at $0.5-3.5\;{\mu}g/ml$ against various Candida albicans strains. The antibiotic showed no cytotoxicity for S180, MKN-45, P388, HeLa and 373 at the concentration of 1 mg/ml. LAM-f4A was colorless powder soluble in water, methanol, ethanol, butanol and negative to ninhydrin reaction. The antibiotic had maximum absorption at 273 nm in methanol, and melting point was $202^{\circ}C$. The molecular weight and formula were determined to be 282 and $C_{14}H_{34}O_5$ by $^1H-NMR,\;^{13}C-NMR$, IR spectrum and elemental analysis.