• Title/Summary/Keyword: HPLC screening

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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.

Determination of Aflatoxin B1 in Rice, Barley, and Feed by Non-instrumental Immunochromatographic Strip-test and High Sensitive ELISA

  • Shim, Won-Bo;Kim, Jung-Sook;Kim, Ji-Young;Choi, Jin-Gil;Je, Jung-Hyun;Kuzmina, Nina Sergeevna;Eremin, Sergei Alexandrovich;Chung, Duck-Hwa
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.623-630
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    • 2008
  • A non-instrumental immunochromatographic (ICG) strip-test and direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for aflatoxin B1 (AFB1) determination were developed and optimized. The detection limits of ICG strip-test and DC-ELISA were 0.5 and 0.004 ng/mL, respectively, and these methods possessed a cross-reaction to aflatoxins. The results of spiked samples by both methods were coincided with the amount spiked AFB1 and the comparative analyses of 172 real samples by 2 immunoassays and high performance liquid chromatography (HPLC) showed a good agreement. Especially, the ICG strip-test is easier to perform and quicker, but less sensitivity than DC-ELISA. Both methods could analyze a high sample throughput with short time, but the sample throughput of ICG strip-test was better. Therefore, the ICG strip-test can be used as a simple, easy, non-instrumental, and fast screening technique for AFB1 determination.

Quantitative Assessment of the Relative Antineoplastic Potential of the n-butanolic Leaf Extract of Annona Muricata Linn. in Normal and immortalized Human Cell Lines

  • George, V. Cijo;Kumar, D.R. Naveen;Rajkumar, V.;Suresh, P.K.;Kumar, R. Ashok
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.699-704
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    • 2012
  • Natural products have been the target for cancer therapy for several years but there is still a dearth of information on potent compounds that may protect normal cells and selectively destroy cancerous cells. The present study was aimed to evaluate the cytotoxic potential of n-butanolic leaf extract of $Annona$ $muricata$ L. on WRL-68 (normal human hepatic cells), MDA-MB-435S (human breast carcinoma cells) and HaCaT (human immortalized keratinocyte cells) lines by XTT assay. Prior to cytotoxicity testing, the extract was subjected to phytochemical screening for detecting the presence of compounds with therapeutic potential. Their relative antioxidant properties were evaluated using the reducing power and $DPPH^*$radical scavenging assay. Since most of the observed chemo-preventive potential invariably correlated with the amount of total phenolics present in the extract, their levels were quantified and identified by HPLC analysis. Correlation studies indicated a strong and significant (P<0.05) positive correlation of phenolic compounds with free radical scavenging potential. The results revealed that the extract was moderately cytotoxic to normal cells with a mean IC50 value of 52.4 ${\mu}g$ when compared with those obtained for cancerous cells (IC50 values of 29.2 ${\mu}g$ for MDA-MB-435S and 30.1 ${\mu}g$ for HaCaT respectively). The study confirms the presence of therapeutically active antineoplastic compounds in the n-butanolic leaf extract of $Annona$ $muricata$. Isolation of the active metabolites from the extract is in prospect.

Gibberellins Production and Identification of Endophytic Fungi Isolated from Aquatic Plant in Fresh Water (담수에 자생하는 수생식물에서 분리된 내생균류의 지베렐린 생산과 동정)

  • You, Young-Hyun;Kang, Sang-Mo;Choi, Yu-Mi;Lee, Myung-Chul;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.43 no.1
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    • pp.71-76
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    • 2015
  • Aquatic plant Hydrocharis dubia (Blume) Backer was collected from the Dalsung wetland in Daegu. Sixteen endophytic fungi with different colony morphologies were isolated from the roots of aquatic plants. Waito-c rice (WR) seedlings were treated with fungal culture filtrates (FCF) for screening plant growth-promoting activity. In the results, HD1008 strain isolated from aquatic plant showed highest plant growth-promoting activity. The FCF of HD1008 strain was analyzed using gas chromatography mass spectrometry (GC/MS) with selected ion monitoring (SIM). Analysis of the FCF of HD1008 strain found that it contained gibberellins (GA) ($GA_1$, 1.2 ng/100 mL; $GA_4$, 5 ng/100 mL). Phylogenetic tree of HD1008 strain was constructed by partial internal transcribed spacer (ITS) region and partial beta-tubulin gene sequences. Therefore, we describe HD1008 strain as a new gibberellin-producing Penicillium trzebinskii based on morphological and molecular characteristics.

Screening and Identification of Fungicidal Compounds Derived from Medicinal Plants against Cucumber Powdery Mildew (약용식물로부터 오이흰가루병에 대한 항균성물질 탐색 및 동정)

  • Paik, Su-Bong;Kyung, Suk-Hun;Doh, Eun-Soo;Oh, Yeon-Sun;Park, Byoung-Keun
    • Korean Journal of Environmental Agriculture
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    • v.13 no.3
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    • pp.301-310
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    • 1994
  • This experiment was conducted to test the fungicidal activity of extracts from 50 medicinal plants to powdery mildew (Sphaerotheca fulinginea) and identify the bioactive substances. Among the medicinal plants tested, the water extract of Rheum undulatum was the most effective in spore germination inhibition, which inhibited by 100% at 200-fold dilution. Also, 50-fold dilution of water extract, 100-fold dilution of alcohol extract, 500-fold dilution of crude extract from Rheum undulatum and even 1000-fold dilution of reference chemical inhibited powdery mildew of cucumber more than 60%. 500-fold dilution of crude extract inhibited powdery mildew of cucumber 100% by twice spray treatment. There was phytotoxcity at the 100-fold dilution, but was not recognized this injury at the 500-fold dilution of crude extract. From our research to identify bioactive substance using HPLC, GLC and Mass spectrum analysis, it indicated that Rheum undulatum extract contained tentatively 1,8-dihydroxy-3-methyl-9,10-anthracenedione and 1,8-dihydroxy-3-methoxy-6-methyl-9,10-anthracenedione.

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Ameliorating Effect of Gardenia jasminoides Extract on Amyloid Beta Peptide-induced Neuronal Cell Deficit

  • Choi, Soo Jung;Kim, Mi-Jeong;Heo, Ho Jin;Hong, Bumshik;Cho, Hong Yon;Kim, Young Jun;Kim, Hye Kyung;Lim, Seung-Taik;Jun, Woo Jin;Kim, Eun-Ki;Shin, Dong-Hoon
    • Molecules and Cells
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    • v.24 no.1
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    • pp.113-118
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    • 2007
  • The brains of Alzheimer's disease (AD) patients are characterized by large deposits of amyloid beta peptide ($A{\beta}$). $A{\beta}$ is known to increase free radical production in nerve cells, leading to cell death that is characterized by lipid peroxidation, free radical formation, protein oxidation, and DNA/RNA oxidation. In this study, we selected an extract of Gardenia jasminoides by screening, and investigated its ameliorating effects on $A{\beta}$-induced oxidative stress using PC12 cells. The effects of the extract were evaluated using the 2',7'-dichlorofluorescein diacetate (DCF-DA) assay and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. To find the active component, the ethanol extract was partitioned with hexane, chloroform, and ethyl acetate, respectively, and the active component was purified by silica-gel column chromatography and HPLC. The results suggested that Gardenia jasminoides extract can reduce the cytotoxicity of $A{\beta}$ in PC 12 cells, possibly by reducing oxidative stress.

A Caspase Inducing Inhibitor Isolated from Forsythiae fructus (연교(Forsythiae fructus)로부터 분리한 caspase 유도 저해물질)

  • Kim, Jin-Hee;Kho, Yung-Hee;Kim, Mee-Ree;Kim, Hyun-A;Lee, Sang-Myung;Lee, Choong-Hwan
    • Korean Journal of Food Science and Technology
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    • v.34 no.1
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    • pp.114-117
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    • 2002
  • During the screening of inhibitors of caspase-3 induction in U937 human monocytic leukemia cells from natural sources, Forsythiae fructus, which showed a high level of inhibition, was selected. And then, the compound was purified from the methanol extract using silica gel column chromatography and HPLC. The inhibitor was identified as rengyolone, by spectroscophic methods of ESI-MS, $^1H-NMR$, $^{13}C-NMR$, DEPT, and HMBC. Rengyolone showed inhibitory activity of caspase-3 induction, a major protease of apoptosis cascade, with an $IC_{50}$ value of $6.25\;{\mu}g/mL$ after 7 h of treatment in U937 cells. It also showed inhibitory activity of caspace-1 induction, with an $IC_{50}$ value of $7.50\;{\mu}g/mL$ after 40 h of treatment in D10S cells. In addition, it showed protective effect against cell death with an $IC_{50}$ value of $11\;{\mu}g/mL$ on U937 cells induced by etoposide after 24 h of treatment, but did not show any cytotoxicity at the same condition without etoposide, a caspase 3 inducing agent.

Screening and Biotransformation of Interleukin-1$\beta$ Converting Enzyme Production Inhibitors from Arctii fructus

  • KIM HYUN A;YOON DO YOUNG;LEE SANG MYUNG;BAEK SEUNG HWA;HAN GYOON HEE;KHO YOUNG HEE;LEE CHOONG HWAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.269-273
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    • 2005
  • Five dibenzylbutyrolactones were isolated from a methanol extract of Arctii fructus (Arctium lappa L.) by bioassay-guided isolation, using the interleukin-l $\beta$ converting enzyme (caspase-l, ICE) production inhibitory assay in vitro. These compounds were spectroscopically identified as lappaol E (1), lappaol A (2), matairesinol (3), arctigenin (4), and arctiin (5). Among the compounds tested, arctigenin (4) showed the strongest inhibitory activity for ICE production in IL-$\beta$-induced proliferation of D 1 OS cells. Western blot analysis demonstrated that the arctigenin suppressed the expression of ICE protein in a dose-dependent manner. To estimate the biotransformation of Arctii fructus in vivo by human intestinal bacteria, we carried out an anaerobic incubation of the Arctii fructus extract with a human fecal suspension. From the HPLC analysis of metabolites, Arctiin (IC$_{50}$=74.2$\mu$g/ml), a major component of Arctii fructus, was transformed to aglycone, arctigenin (IC$_{50}$=12.5$\mu$g/ml), by human intestinal bacteria. The ICE production inhibitory activity of Arctii fructus would be much stronger in vivo than in vitro due to the biotransformation by human intestinal bacteria.

Pyridoxatin, an Inhibitor of Gelatinase A with Cytotoxic Activity

  • Lee, Ho-Jae;Chung, Myung-Chul;Lee, Choong-Hwan;Chun, Hyo-Kon;Kim, Hwan-Mook;Kho, Yung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.445-450
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    • 1996
  • Gelatinase A is a member of the matrix metalloproteinases that play an important role in cancer invasion and metastasis. In the course of screening gelatinase A inhibitors from microbial sources, a fungal strain PT-262 showed a strong inhibitory activity. The strain was identified as Chaunopycnis alba on the basis of its morphological characteristics. The inhibitor was isolated from acetone extract of mycelial cake by sequential chromatographies on MCI-gel, Sephadex LH-20, and a reverse-phase HPLC column. The purified inhibitor was identified as pyridoxatin by its physico-chemical properties and spectroscopic analysis. Pyridoxatin is not a peptide analog and has cyclic hydroxamic acid moiety. It inhibited activated gelatinase A with an $IC_{50}$ value of 15.2 ${\mu}M$ using fluorescent synthetic peptide. It also had a strong cytotoxicity against human cancer cell lines in vitro. Furthermore, this compound inhibited DNA synthesis with an $IC_{50}$ value of 2.92 ${\mu}M$ in PC-3 prostate cancer cells by [$^3H$]thymidine incorporation assay.

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Insecticidal Activities of Tussilago farfara Extracts against Culex pipiens pallens and Tetranychus urticae (빨간집모기와 점박이응애에 대한 관동화(Tussilago farfara) 추출물의 살충효과)

  • Park, Sung Ho;Oh, Hyun-Woo;Kwon, Hye-Ri;Seo, Mi-Ja;Yu, Yong-Man;Youn, Young-Nam
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.177-185
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    • 2014
  • Aspiring to the new raw materials of insecticides is one of the plant extracts. The material structure of a variety of plant extracts have because the material to the defense of the plant itself, the case of insecticides, using plant extracts, safe, has low toxicity and has the advantage of highly distinctive and fall. Coltsfoot (Tussilago farfara L.) is belonging to the family Compositae, and distributed in all parts of China as medicinal plants and indigenous plants used. Coltsfoot is known that it is effect to respiratory disease and has an antiviral effect. However, the reported insecticidal activity of coltsfoot could be not found. Fortunately, I found insecticidal activities when I was screening the bioassay against several insects with a lot of plant extracts. Using the ethanol extract of the Tussilago farfara, there were insecticidal activities against Culex pipiens pallens and Tetranychus urticae. There were several fractions in ethanol extract of coltsfoot by using various organic solvents. Hexane fraction showed a higher insecticidal activity than any other fraction. It is confirmed that hexane fraction contained pyrethrin by using HPLC analysis. So, it might be suggested that extract of coltsfoot has an insecticidal activity and its effect due to a ingredient of pyrethrin.