• Title/Summary/Keyword: silica-gel column chromatography

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Isolation an Identification of the Active Compounds from Green Mustard Leaves against Oxidative Stress in Bovine Brain Tissue (청겨자채 추출물의 뇌조직 내 Oxidative Stree 억제활성물질의 동정)

  • 이근종;김미리;심재석;황재관
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.943-948
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    • 2001
  • Green mustard leves were found to effectively prevent lipid peroxidation of bovine-brain tissue by ascor-bate/Fe system, The 50% methanol extracts mustard leaves were separated into four solvent faction using n-hexane,. EtOAc, n-BiOH and water. Then n-BiOH fraction exclusively exhibited the antioxidative activities at concentration above 100 $\mu\textrm{g}$/mL/ The n-BuOH fraction was further isolated to a single compound using TLC analysis and silica gel chromatography. The active antiodidative compounds were identified as sinapic acid methyl ester and ferulic acid methyl ester by $^{1}$H-NMR and $^{13}$ C-NMR, The sinapic acid methyl ester and ferulic acid methyl ester were prepared by methylating of sinapic acid and ferulic acid with diazomethane. The results strongly suggested that sinapic acid and ferulic acid could be emplyed as a potential antioxiative agents for preventing the bovine brain lipid peroxidation. lipid peroxidation.

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Development of Biologically Active Compounds from Edible Plant Sources XXII. Triterpenoids from the Aerial Parts of Sajabalssuk (Artemisia princeps PAMPANINI) (식용식물자원으로부터 활성물질의 탐색-XXII. 사자발쑥(Artemisia princeps PAMPANINI)의 지상부로부터 Triterpenoid의 분리)

  • Bang, Myun-Ho;Cho, Jin-Gyeong;Song, Myoung-Chong;Lee, Dae-Young;Han, Min-Woo;Chung, Hae-Gon;Jeong, Tae-Sook;Lee, Kyung-Tae;Choi, Myung-Sook;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.223-227
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    • 2008
  • The aerial parts of Sajabalssuk (Artemisia princeps PAMPANINI, Sajabalssuk) was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and $H_2O$, successively. From the EtOAc fraction, three cycloartane-type triterpnoids and one ursane-type triterpenoid were isolated through the repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the results of physico-chemical data including NMR, MS and IR, the chemical structures of the triterpenoids were determined as wrightial (1), wrightial acetate (2), 27-norcycloart-20(21)-ene-25-al-3${\beta}$-ol acetate (3) and ursolic acid (4). No report has been found for isolation of compound 3 in the literature so far, and compounds 1, 2 and 3 were the first to be isolated from Sajabalssuk (Artemisia princeps PAMPANINI, Sajabalssuk). Also, compound 1 showed Acyl-CoA:Cholesterol acyltransferase (hACAT-1) and hACAT-2 inhibitory activity with the $IC_{50}$ values of 33.0 and 45.0 ${\mu}g/ml$, respectively. Compounds 2 and 3 inhibited hACAT-1 activity with the $IC_{50}$ values of 12.0 and 16.0 ${\mu}g/ml$, respectively.

Preparation of minor ginsenosides C-Mc, C-Y, F2, and C-K from American ginseng PPD-ginsenoside using special ginsenosidase type-I from Aspergillus niger g.848

  • Liu, Chun-Ying;Zhou, Rui-Xin;Sun, Chang-Kai;Jin, Ying-Hua;Yu, Hong-Shan;Zhang, Tian-Yang;Xu, Long-Quan;Jin, Feng-Xie
    • Journal of Ginseng Research
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    • v.39 no.3
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    • pp.221-229
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    • 2015
  • Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

Antioxidative compounds of Achillea sibirica Ledeb (톱풀의 항산화 성분)

  • Moon, Hyung-In;Lyu, Sung-Hyo;Roh, Jong-Hwa;Zee, Ok-Pyo
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.1
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    • pp.1-8
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    • 2000
  • Achillea sibirica Ledeb. is widely distributed in Korea and has been often used as folk medicine in peptic and tonic. As one of our searches for bioactive (anti oxidation) compounds from medicinal plants, we studied Achillea sibirica Ledeb. (Compositae). Antioxidant activity of Achillea sibirica was determined by measuring lipid peroxide produced when a mouse liver homogenate was exposed at $90^{\circ}C$ using 2-thiobarbituric acid (TBA) and by evaluation the radical scavenging activity on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical. Whole parts of Achillea sibirica was extracted with methanol and its extracts was fractionated with organic solvent; n-Hexane, methylene chloride, ethyl acetate, n-Butanol. EtOAc fraction exhibited antioxidant activity and From its, two flavonoid glycosides were isolated by silica gel and gel filtration colume chromatography and identified to kampferol 3-O-glucoside and luteolin 7-O-neo-hesperidoside, respectively, by physico-chemical and spectroscopical method. At antioxidant activity test for two compounds isolated, antioxidant activity was showed too. And from hexane fraction sterol was is isolated and identificated to mixture of campesterol, stigmasterol, and ${\beta}-sitosterol$.

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Isolation and Identification of Antioxidant Substances from the Stems of Butterbur (Petasites japonicus) (머위(Petasites japonicus) 엽병으로부터 항산화 물질의 분리 및 동정)

  • Kim, Min-Young;Yi, Jung-Hyun;Hwang, Yun-Yi;Song, Kyung-Sik;Jun, Mi-Ra
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.8
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    • pp.979-984
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    • 2008
  • The stems of P etasites japonicus were extracted with ethanol and then partitioned with hexane, chloroform, ethyl acetate, n-butanol and water, successively. The antioxidant potency of five crude fractions were determined using (1) 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, (2) thiobarbituric acid reactive substances (TBARS) assay in the linoleic acid model system, and (3) lipoxygenase inhibition assay. Among the crude fractions, the ethyl acetate fraction exhibited the most potent antioxidant effect. By activity-guided fractionation, compound PJ-4 was isolated from the ethyl acetate fraction through the repeated silica gel open column chromatography. The chemical structure of the isolated compound was determined as kaempferol by $^1H-$ and $^{13}C$-NMR analysis and its antioxidative capacity was further investigated. DPPH radical scavenging activity of the compound was 65.76% at the concentration of $100 \;{\mu}g/mL$. The inhibitory activity of the compound against lipid peroxidation and lipoxygenase exhibited 43.47% and 58.60%, respectively at the concentration of $100\;{\mu}g/mL$. The result suggests that the compound may serve as a useful natural antioxidant and furthermore indicates the possibility of developing the stems of Petasites japonicus as a natural antioxidant source.

Catechin Content and Antioxidative Effect from Rosa davurica Pall (생열귀나무의 카테킨 함량 및 항산화효과)

  • Shin, In-Cheol;Jeong, Kyung-Jin;Shim, Tae-Heum;Oh, Heung-Seok;Park, Sang-Kyun;Cheung, Eui-Ho;Kim, Suk-Nam;Kim, Gwang-Gee;Choi, Dae-Sung;Kwon, Yong-Soo;Kim, Chang-Min;Sa, Jae-Hoon
    • Korean Journal of Pharmacognosy
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    • v.33 no.3 s.130
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    • pp.177-181
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    • 2002
  • This study was carried out to investigate the antioxidative activities of Rosa davurica Pall for the purpose of development of novel antioxidant from natural products. Antioxidant activities of four different parts of Rosa davurica Pall such as fruit, leaf, stem and root were examined by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. The methanol extract from the root of Rosa davurica Pall showed the highest antioxidative activity among 16 samples tested. And, we also tested radical scavenging effects of 5 different extract compartments(Hexane, $CHCl_3$, EtOAc, BuOH and $H_2O$ fraction). EtOAc and BuOH fractions from the root of Rosa davurica Pall exhibited antioxidative activities higher to those of natural, ${\alpha}-tocopherol$ or synthetic antioxidants, BHT. The antioxidative substance of EtOAc fraction from the root of Rosa davurica Pall was successively purified with silica gel adsorption column chromatography and Sephadex LH-20 column chromatography. The purified active substance was isolated as crystal and identified as (+)-catechin by $^{l}H-NMR$ and $^{13}C-NMR$. This compound exhibited DPPH radical scavenging activity with the $IC_50$ value of $1.7\;{\mu}g/ml$. In the analysis of catechin content, the leaf extracts contained the highest catechin, and fruit extracts contained the lowest catechin. Considering antioxidative activity on DPPH assay, the extracts of Rosa davurica Pall showed a possibility to be used as a new material for natural antioxidant and functional food.

Isolation, Identification and Characterization of Paraquat Activity-Inhibiting Substances in Squash (Cucurbita moschata Duchesne ex Poiret) Leaves (호박잎에서 Paraquat 활성 억제 물질의 분리, 동정 및 특성 구명)

  • Hyun, Kyu-Hwan;Yun, Young-Beom;Jang, Se-Ji;Shin, Dong-Young;Kwon, Oh-Do;Choi, Hyun-Sug;Jung, Ha-Il;Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.32 no.3
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    • pp.211-221
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    • 2012
  • The fourth leaves (younger leaves) amongst extended 4-upper leaves in 18 squash cultivar were the highest tolerance to the paraquat application, followed by third, the second, and the first leaves (older leaves). The forth leaves in Joongangaehobak showed more than three times higher tolerance to the paraquat application than did the first leaves. When the combining of water extract from the fourth leaves with paraquat were applied to the leaves and stems of maize, the paraquat phytotoxicity in maize was reduced compared to the paraquat application alone. Therefore, this study continued to investigate if the phytotoxicity inhibitor exist in the fourth leaves. The water extract in the fourth leaves were isolated by silica gel column chromatography, Sephadex LH-20 column chromatography, TLC, and HPLC, and the substance in the extract was speculated as a malic acid by identifying through NMR. The mixture malic acid and paraquat were applied to the maize to verify the application effect of malic acid on paraquat toxicity. The 100 ${\mu}M$ of paraquat application alone showed 62% of paraquat toxicity to the corn leaves, while the combined application of 100 ${\mu}M$ paraquat with malic acid at 0.1, 0.3, 0.5, and 1.0% did not show the symptom.

Synthesis of Substrates for Gene Therapy Monitoring of HSV1-TK System (유전자 영상용 HSV1-TK 기질의 합성)

  • Choi, Tae-Hyun;Ahn, Soon-Hyuk;Choi, Chang-Woon;Lim, Sang-Moo;Awh, Ok-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.2
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    • pp.102-109
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    • 2002
  • Purpose : In gene therapy, tumor cells expressing the herpes simplex virus thymidine kinase are sensitive to prodrugs. Potential prodrugs IVDU and IVFRU were synthesized and radiolabeled with radioiodine for noninvasive imaging of herpes simplex virus type 1 gene expression. Material and Methods : 5-(2-trimethysilyl) vinyl-2'-deoxyuridine and 5-(2-trimethylsilyl)vinyl-2'-fluoro-2'-deoxyuridine, precursors of 5-(2--iodo)viny l-2'-deoxy uridine(IVDU) and 5-(2-iodo)-2'-vinyl-2'-deoxy-2'-fluororibofuranosyl uracil(IVFRU), were synthesized from reaction of trans-1-trimethylsillyl-2-tri-n-butylstannylethylene with 5-iodo-2'-deoxyuridine and 5-iodo-2'-fluoro-2'-deoxyuridine, respectively, on the condition of Pd catalyst. These precursors were separated from reaction mixture by silica gel column chromatography method. Each precursor was radioiodinated with radioiodine by mixing with ICI oxidizing agent. These radioiodinated compounds were purified with HPLC. Radiohalogen exchange has been shown to be effective for the synthesis of products with lower specific activity. Similarly, carrier-added and high specific activity products have been isolated in respectable radiochemical yields using ICI method. Results : Synthetic yield of precursors, IVDU and IVFRU were 43% and 18%, respectively. Radiochemical purity of both compunds was over 98%. Conclusion : We synthesized precursors of IVDU and IVFRU for monitoring of HSV1-tk gene expression. Radiotracers were radioiodinated with high radiolabeling yield by ICI method.

In vitro Cytotoxin Activity of Urushiol in the Sap of Rhus verniciflua STOKES (옻나무 칠액성분(漆液成分) 중 Urushiol의 암세포(癌細胞) 증식억제(增殖抑制) 효과(效果) - in vitro 세포독성효과(細胞毒性效果) -)

  • Na, Chun-Soo;Jung, Nam-Chul;Oh, Kwang-In
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.260-269
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    • 1998
  • This study was conducted to screen the biological activity of urushiol in the sap of lac tree(Rhus verniciflua STOKES) which has been used in traditional folk remedies. Cytotoxic activity of urushiol was screened with L1210(mouse luekemia cell), PC-9(human lung adenocarcinoma cell), A427(human lung adenocarcinoma cell) and KATO III (human stomach adenocarcinoma cell) The stepwise hexane : acetone eluent fractions of the urushiol were obtained by the silica gel adsorption column chromatography and added to the culture media containing L1210, PC-9. A427, and KATO III, respectively. A hexane : acetone(90 : 10, v/v) eluent fraction of them showed the lowest 50% inhibition concentration($IC_{50}$) of $0.018{\mu}g/m{\ell}$ for the cell line of A427. Much lower level of $IC_{50}$ of the hexane : acetone(90 : 10, v/v) eluent fraction of the urushiol showed the equal inhibition effect with tetraplatin(i.e., anti-cancer drug of platinum complexes) on the cancer cell lines as follows ; 3.4 times lower for L1210, 3.9 times lower for PC-9, and 105.5 times lower for A427. However, $IC_{50}$ of the hexane : acetone(90 : 10 v/v) eluent fraction for KATO III was exceptionally 3.9 times higher than that of tetraplatin.

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Studies on the Cellulase. (V) -Fractionation of Cellulolytic Complex produced by Trichoderma $viride(O_2-1)$ (섬유소(纖維素) 분해효소(分解酵素)에 관(關)한 연구(硏究) (제5보(第5報)) -Trichoderma $(O_2-1)$가 생성(生成)하는 Cellulolytic Complex의 분별(分別)에 대(對)하여-)

  • Sung, Nack-Kie
    • Applied Biological Chemistry
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    • v.12
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    • pp.99-105
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    • 1969
  • The yield of cellulase derived from Trichoderma $(O_2-1)$ was remarkably varied with various concentration of ethanol and acetone in purification of the enzyme. In the purification with ethanol of ${\beta}-glucosidase$, the best result was obtained in the concentration of 60% and, of CMCase and of filter paper disintegrating enzyme 80%. And in the purification with acetone of ${\beta}-glucosidase$, filter paper disintegrating enzyme, and CMCase, in the concentration of 60%, 80%, and 90% respectively, was shown the best yield. The activities of crude Cellulase preparation could be seperated into few of fractions by column chromatography with Silica gel, Cellulose powder, and gauze. Most of CMCase, avicelase, and ${\beta}-glucosidase$ were eluted, but most of filter paper disintegrating enzyme and the rest of enzymes mentioned the above were absorbed, and were eluted with water. Therefore, it was considered that CMCase is different from filter paper disintegrating enzyme in properties. The relative activity of CMCase was different from that of avicelase in the peak of elusion part. And it was considered that filter paper disintegrating enzyme and cellulose powder saccharifying enzyme was seperated respectively as absorption part and non absorption part. The auther came to the conclusion that at least there were more than three sorts of cellulase in Trichoderma $(O_2-1)$ cellulase preparation.

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