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

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Biological Activities and Bioactive Compounds in the Extract of Acer tegmentosum Maxim. Stem (산겨릅나무 줄기추출물의 생리활성 및 유효성분 분리)

  • Hong, Bo-Kyong;Eom, Seok-Hyun;Lee, Chan-Ok;Lee, Ji-Won;Jeong, Jong-Hyun;Kim, Jae-Kwang;Cho, Dong-Ha;Yu, Chang-Yeon;Kwon, Yong-Soo;Kim, Myong-Jo
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.296-303
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    • 2007
  • Acer tegmentosum (Acereaceae) has been used a source of traditional medicines for the treatment of hepatic disorders in Korea. This research was conducted to determine biofunctional activities of A. tegmentosum stem extract and to identify its bioactive components. Methanolic extract from A. tegmentosum stem was partitioned by using organic solvents, including n-hexane, ethyl acetate, n-butanol, and water. Two compounds were isolated by using an ODS column chromatography from ethyl acetate soluble fraction shown to the strongest antioxidant activity ($RC_{50}=3.15\;{\mu}g/m{\ell}$) among the fractions. The isolated compounds were analyzed by $^1H$ and $^{13}C$ NMR, IR, UV/VIS, MS spectrum data and identified as catechin, ${\rho}-Hydroxyphenethyl$ alcohol $1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$. The compounds have shown strong antioxidant activity, with similar activity to BHA ($RC_{50}=2\;{\mu}g/m{\ell}$). Especially, ${\rho}-Hydroxyphenethyl$ alcohol 1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$ was shown strong anti-lipid peroxidative activity. However, the compounds were not shown antimicrobial activities. In antimicrobial activity assays, ethyl acetate soluble fraction was effective to bacterial inhibition, such as Escherichia coli and Klebsiella pneumonia, with minimum inhibitory concentrations in $125\;{\mu}g/m{\ell}$. Otherwise, antifungal activity against Candida albicans was shown in n-hexane soluble fraction exhibiting $63\;{\mu}g/m{\ell}$ of minimum inhibitory concentration. In anticomplementary activity assays, water soluble fraction was the most effective exhibiting 24% inhibitory activity.

Purification of Antimicrobial Compounds and Antimicrobial Effects of Schima wallichii subsp. liukiuensis against Candida sp. (Schima wallichii subsp. liukiuensis의 Candida종에 대한 항균효과 및 항균물질의 분리정제)

  • Choi, Mynug-Suk;Shin, Kuem;Yang, Jae-Kyung;Ahan, Jin-Kwon;Kwon, Oh-Woong;Lee, Yi-Young
    • KSBB Journal
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    • v.16 no.3
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    • pp.269-273
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    • 2001
  • To develop natural antimicrobial substances from Theaceae, Schima wallichii subsp. liukiuensis was selected from 218 woody plants, and antimicrobial compounds against bacteria, fungi, and yeast were isolated. The antimicrobial activity of ethanol extracts proved higher than those of other organic solvents. The antimicrobial activity of S. liukiuensis extract showed no differences in sesonal variation, but, that of plant part was high in bark at autumn. An antimicrobial substance was isolated from the extract of Schima using column chromatography packed with silica gel and sephadex LH-20, and then a purified antimicrobial substance (Compound I) was obtained using HPLC analysis. The Compound I in the analysis of UV, IR, and GC-MS presumed a triterpene or steroidal saponin, ${\alpha}$-sitisterol as aglycon combined three sugars. The minimal inhibitory concentration (MIC) of the Compound I against a bacteria, fungi, and yeast were 1.25 g/L, 5.0 g/L, and 0.040 g/L, respectively. This is much lower than the MIC of hinokitiol, an natural antimicrobial compound used commercially, which suggests that Compound I could be developed as a natural preservative and pharmaceuticals.

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Nitrite Scavenging Ability and SOD-like Activity of a Sterol Glucoside form Chrysanthemum coronarium L. var. spatiosum (쑥갓 스테롤배당체의 아질산염소거작용 및 SOD 유사활성)

  • Cho, Min-Jung;Park, Mi-Jung;Lee, Heum-Sook
    • Korean Journal of Food Science and Technology
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    • v.39 no.1
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    • pp.77-82
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    • 2007
  • From the total methanolic extract of Chrysanthemum coronarium L. var. spatiosum (Compositae), nitrite scavenging ability and superoxide dismutase (SOD)-like activity were analyzed as antioxidative characteristics. After successive partitioning with chloroform, n-butanol, and water, the chloroform fraction showed the most significant nitrite scavenging ability with an $IC_{50}$ value of 39 ppm compared with the values of vitamin C and chlorogenic acid, 15 ppm and 36 ppm, respectively. The active fraction was subjected to silica gel and Sephadex LH-20 column chromatography, and the compound was isolated and identified as ${\beta}-sitosterol-O-{\beta}-D-glucoside$ using $^{1}H-NMR$ and $^{13}C-NMR$ spectral data. The glucoside was further hydrolyzed and confirmed as a glycosylated ${\beta}-sitosterol$. The compound and its aglycone, ${\beta}-sitosterol$, showed different nitrite scavenging and SOD-like activity. The $IC_{50}$ value of nitrite scavenging ability of the compound was 335 ppm at pH 1.5, while that of its aglycone was 41 ppm. As for the SOD-like activity, the $EC_{50}$ values of the sterol and the glucoside were 1,291 ppm and >2,000 ppm, respectively, compared with those of vitamin C and chlorogenic acid, 38 ppm and 449 ppm, respectively.

Development of Analytical Method for Cymoxanil in Agricultural Commodities using HPLC/UVD (HPLC/UVD를 이용한 농산물 중 cymoxanil의 개별 분석법 확립)

  • Kim, Ji-Yoon;Kim, Hea-Na;Kim, Ja-Young;Kim, Jong Geol;Ham, Hun-Ju;Lee, Young-Deuk;Hur, Jang-Hyun
    • The Korean Journal of Pesticide Science
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    • v.18 no.2
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    • pp.69-78
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    • 2014
  • In the present study, we developed an official individual analytical method for cymoxanil using HPLC/UVD, respectively in different representative crops. Individual analytical methods for these pesticides are not included in the Korea food code. The samples were extracted with acetonitrile, concentrated and partitioned with dichloromethane and saturated sodium chloride solution. For cymoxanil, extracts were concentrated and clean-up through silica gel column chromatography with dicloromethane/acetone (60/40 v/v) and subjected to instrumental analysis. The limit of detection (LOD) for cymoxanil were 0.1 ng and 1 ng respectively and limit of quantitation (LOQ) were 0.02 mg/kg. Recoveries for cymoxail ranged from 79.6~107.6% respectively, at fortification level of 0.02 mg/kg (LOQ), 0.2 mg/kg (10 LOQ) and 1.0 mg/kg (50 LOQ) and the coefficient of variation (CV) was less than 10%, regardless of sample types. These results were further confirmed with LC/MS. The proposed simultaneous analysis method is reproducible and sensitive enough to determine the residues of cymoxanil in the agricultural commodities. According to the validation data and performance characteristics and high sample throughput, the proposed method is suitable for routine application.

Prolyl Endopeptidase-inhibiting Isoflavonoids from Puerariae Flos and Some Revision of their $^{13}C-NMR$ Assignment (갈화의 Prolyl Endopeptidase 저해 활성 Isoflavonoid 및 이들의 $^{13}C-NMR$ Assignment)

  • Kim, Kyung-Bum;Kim, Sang-In;Kim, Jong-Sik;Song, Kyung-Sik
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.351-355
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    • 1999
  • In order to find anti-dementia drugs from natural products, prolyl endopeptidase inhibitors were purified from Puerariae Flos by consecutive solvent partition, followed by silica gel, Sephadex LH-20, and HPLC. Four isoflavonoid inhibitors were isolated and identified as tectorigenin, genistein, 5,7-dihydroxy-4',6-dimethoxyisoflavone, and 5-hydroxy-6,7,4'-trimethoxyisoflavone by means of instrumental analyses including $^{1}H-$, $^{13}C-$, $^{2}D-NMR$ and MS and $IC_{50}$ values against PEP were 5.30 ppm$(17.7\;{\mu}M)$, 10.39 ppm$(38.5\;{\mu}M)$, 13.92 ppm$(44.3\;{\mu}M)$, and 20.61 ppm$(62.8\;{\mu}M)$, respectively. Some previous mistakes in $^{13}C-NMR$ assignment were revised by careful investigation of HMBC and HMQC data.

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Inhibition of Low Density Lipoprotein-oxidation, ACAT-1, and ACAT-2 by Lignans from the Bark of Machilus thunbergii

  • Shrestha, Sabina;Park, Ji-Hae;Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Do-Gyeong;Cho, Moon-Hee;Jeong, Tae-Sook;Kang, Hee-Cheol;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.63-66
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    • 2011
  • The bark of Machilus thunbergii was extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc fraction, five lignans were isolated through the repeated silica gel, octadecyl silica gel (ODS) and, Sephadex LH-20 column chromatography. Based on nuclear magnetic resonance (NMR), mass spectroscopy (MS), and infrared spectroscopy (IR) spectroscopic data, the chemical structures of the compounds were determined to be machilin A (1), machilin F (2), licarin A (3), nectandrin A (4), and nectandrin B, (5). This study presents comparative account of five lignans from M. thunbergii bark contributing inhibition of low density lipoprotein (LDL), ACAT-1, and ACAT-2. Compounds 2-5 showed varied degree of antioxidant activity on LDL with $IC_{50}$ values of 2.1, 11.8, 15.3, and $4.1{\mu}M$. Compounds 1, 2, and 3 showed inhibition activity on ACAT-1 with values $63.4{\pm}6.9%$ ($IC_{50}=66.8{\mu}M$), $53.7{\pm}0.9%$ ($IC_{50}=109.2{\mu}M$), and $78.7{\pm}0.2%$ ($IC_{50}=40.6{\mu}M$), respectively, at a concentration of 50 mg/mL, and on ACAT-2 with values $47.3{\pm}1.5%$ ($IC_{50}=149.7{\mu}M$), $39.2{\pm}0.2%$ ($IC_{50}=165.2{\mu}M$), and $52.1{\pm}1.0%$ ($IC_{50}=131.0{\mu}M$, respectively, at a concentration of 50 mg/mL.

Screening and Purification of an Anti-Prostate Cancer Compound, Deoxypodophyllotoxin, from Anthriscus sylvestris Hoffm (전호(Anthriscus sylvestris Hoffm)로부터 전립선 암세포 저해물질인 deoxypodophyllotoxin 의 탐색 및 분리)

  • Cho, Hyo-Jin;Yu, Sun-Nyoung;Kim, Kwang-Youn;Sohn, Jae-Hak;Oh, Hyun-Cheol;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.19 no.1
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    • pp.9-14
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    • 2009
  • The prostate cancer is the critical health problem, increasing of its related death in worldwide. Unfortunately present surgery and chemotherapeutic choices seem to be impossible in curing or controlling prostate cancer, because metastasis occasionally advances even after these potentially curative therapies. Therefore, there is immediate need to alternative chemoprevention and chemotherapeutic agents. Over one hundred species of dried medicinal herbs were tested for proliferation inhibitory effects on prostate cancer cell line, PC-3. One of them, Anthriscus sylvestris was selected because of potent anti-proliferation effect. The dried root of A. sylvestris was extracted with 100% methanol for 2-3 days and its extract was fractionated by using ethyl acetate. And ethyl acetate layer was subjected to column chromatographies on silica gel, reverse phase-18 (RP-18) and Sephadex LH-20, in turn. Finally, the pure compound was obtained by crystallization in methanol at $4^{\circ}C$ for overnight and identified as deoxypodophyllotoxin by NMR spedorscopic and physico-chemical analyses. In addition, it was confirmed that deoxypodophyllotoxin clearly inhibits the proliferation of PC-3 cells in a dose and time-dependent manner.

Antimicrobial Activity of the Coriolus versicolor Liquid Culture Extracts Against Antibiotic Resistant Bacteria and Purification of Active Substance (구름버섯 균사체 배양 추출물의 복합내성 세균에 대한 항균활성 및 활성물질의 정제)

  • Lee, Jung-Sun;Kim, Taeg;Lee, Yoon-Hi;Jin, Cheng-Min;Kim, Hyun-Guell;Kim, Woo-Jung;Oh, Duek-Chul;Park, Yong-Il
    • The Korean Journal of Mycology
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    • v.34 no.2
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    • pp.92-97
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    • 2006
  • The liquid culture extract of Coriolus versicolor was prepared by directly boiling the whole culture broth 7 days after incubation in 12% citrus extract medium. After removal of mycelial debris through filtration, this extract was further extracted with equal volume of ethyl acetate (1 : 1, v/v). The ethyl acetate extracts showed significant antibacterial activities against Stapylococcus aureus CCARM3230 and Psudomonas aeruginosa CCARM2171, which are resistant to several antibiotics. The most active fraction was eluted from a silica gel column with a mixture of dichloromethane and methanol (9 : 1, v/v) and the purity of this active substance was confirmed by HPLC analysis. The results suggest that the purified active substance could be a good source for the development of a new antimicrobial agent, especially for the treatment of antibiotic resistant bacteria.

Gas Chromatographic Analysis on the Residual of Fungicide Fenhexamid in Crops(Cucumber, Strawberry and Grape) (작물(오이, 딸기, 포도) 중 살균제 Fenhexamid의 잔류성에 대한 기체 크로마토그래피 분석)

  • Han, Seong-Soo;Lo, Seog-Cho;Kim, Won-Ju;Park, PiII-Jae;Kim, Il-Kwang
    • Analytical Science and Technology
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    • v.16 no.1
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    • pp.70-77
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    • 2003
  • The optimum conditions for the residue analysis of hydroxyanilide fungicide fenhexamid on cucumber, strawberry and grape were investigated by gas chromatography equipped with electron capture detector (GC-ECD) and the residual amount was determined by sprayed days before harvest. Each samples were extracted with acetone, filtered and concentrated to 50 mL. The concentrated extracts were transferred to dichloromethane and then thoroughly concentrated. The concentrated phase was loaded on the filtration column stuffed with silica gel and purified with acetone:hexane (5:95, 15:85, v/v) mixed solvent. The regression equation and linearity of the standard calibration curves between 0.05~2.00 ng were as follows : cucumber; Y=312.40X+10.26, $R^2=0.9996$, strawberry; Y=313.33X+5.54, $R^2=0.9998$, grape; Y=253.27X-2.23, $R^2=0.9994$. From the standard additional experiments with 0.10 mg/L and 0.40 mg/L, the average recoveries of cucumber, strawberry and grape were 94.8%, 88.1% and 93.7%, respectively and the detection limits were all the same as 0.01 mg/L. Residual amounts in crops were ranged from 0.01 to 0.58 mg/L.

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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