• 제목/요약/키워드: HPLC screening

검색결과 159건 처리시간 0.028초

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

  • 조효진;유선녕;김광연;손재학;오현철;안순철
    • 생명과학회지
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    • 제19권1호
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    • pp.9-14
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    • 2009
  • 전립선암은 현대 남성들에게 걸리기 쉬운 질병으로 나이가 들수록 발병의 위험이 증가하는 질환으로 현재 우리나라에서도 점점 증가하는 추세이다. 전립선암 치료법들은 치료영역이 제한적이고 재발할 가능성이 높아 근본적인 치료법으로는 사용되지는 못하므로 새로운 전립선암 치료 방법이 필요하다. 이에 본 연구에서는 100 여 가지의 한약재 methanol 추출물을 이용하여 MTT 방법으로 전립선암 세포주인 PC-3 세포에 대한 항증식 효과를 탐색하였으며 그 결과, A. sylvestris가 가장 강한 항증식 활성을 보였다. A. sylvestris의 methanol 추출물로부터 저해물질을 분리하기 위하여 100% methanol에서 2-3일 추출하고 난 뒤, ethyl acetate로 추출하고 silica gel, reverse phase-18, Sephadex LH-20 등의 컬럼 크로마토그래피를 이용하여 분리하였다. 최종적으로 활성분획을 HPLC로 분리하고 $4^{\circ}C$에서 methanal 용액에서 입방체 형태의 결정을 얻었으며 NMR 분광법과 이화학적 특성을 분석한 결과, deoxypodophyllotoxin 으로 동정되었다. 순수 분리된 deoxypodophyllotoxin은 전립선암의 세포주 PC-3 세포에서 처리 농도와 처리 시간 의존적인 항증식 효과를 보였다.

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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    • 제17권3호
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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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    • 제13권2호
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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.

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

  • 유영현;강상모;최유미;이명철;김종국
    • 한국균학회지
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    • 제43권1호
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    • pp.71-76
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    • 2015
  • 수생식물 샘플은 낙동강과 금호강이 만나는 달성습지에서 자라풀을 채집하였다. 자라풀의 뿌리에서 내생균류를 분리하고 형태가 다른 균주를 관찰하여 최종적으로 16개 균주를 선발하였다. 내생균류의 배양여과액은 식물생장촉진활성 검정을 위하여 난장이벼에 처리하여 스크리닝하였으며, HD1008 균주가 식물생장촉진활성이 가장 높은 것으로 확인되었다. HD1008 균주의 배양여과액을 HPLC와 GC/MS-SIM을 이용하여 분석하였고, HD1008 균주가 식물호르몬인 지베렐린 $GA_1$ (1.2 ng/100 mL)과 $GA_4$ (5 ng/100 mL) 를 생산하는 것을 정량분석을 통하여 확인하였다. 또한, HD1008균주의 beta-tubulin 유전자 염기서열을 이용하여 동정에 이용하였으며, 분자적인 방법과 형태적인 방법으로 관찰하였을 때, 지베렐린을 생산하는 새로운 P. trzebinskii로 동정되었다.

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

  • 백수봉;경석헌;도은수;오연선;박병근
    • 한국환경농학회지
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    • 제13권3호
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    • pp.301-310
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    • 1994
  • 50종의 약용식물추출액을 공시하여 흰가루병에 대한 방제효과를 검정하고 생리활성 물질을 분리, 동정하였다. 공시약용식물중에서 대황의 물추출액은 200배의 희석농도에서도 100%의 포자발아억제 효과를 나타내서 가장 효과가 있었고, 대황의 물추출액 50배, 알콜추출액 100배, 조물질 500배, 표준품 1000배 희석농도까지도 60%이상의 발병억제효과가 있었으며, 조물질 500배액에서 2회 이상 살포하면 100% 발병억제 효과가 있었다. 그리고 조물질 100배 액에서는 약해가 있었으나 500배액에서는 약해가 없었다. 대황에 함유되어 있는 생리활성물질은 anthraquinone유도체인 1,8-dihydroxy-3-methyl-9,10-anthracenedione과 1,8-dihydroxy-3-methoxy-6-methyl-9,10-anthracenedione으로 잠정 동정되었다.

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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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    • 제24권1호
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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.

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

  • 김진희;고영희;김미리;김현아;이상명;이충환
    • 한국식품과학회지
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    • 제34권1호
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    • pp.114-117
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    • 2002
  • 연교의 메탄올 추출물로부터 caspase 저해물질을 분리하였으며, ESI-MS, $^1H-NMR$, $^13C-NMR$, DEPT 등의 기기분석 자료에 의하여 rengyolone으로 동정하였다. 이 물질은 $IC_{50}\;6.25\;{\mu}g/mL$의 농도에서 etoposide가 처리된 U937 세포주의 caspase-3 유도 저해를 나타내었다. 또한 rengyolone은 $Interleukin-1{\beta}$가 처리된 D10S 세포에서 caspase-1의 유도저해활성을 나타내었으며, $IC_{50}$값은 $7.5\;{\mu}g/mL$이었다.

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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    • 제15권2호
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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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    • 제6권6호
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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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빨간집모기와 점박이응애에 대한 관동화(Tussilago farfara) 추출물의 살충효과 (Insecticidal Activities of Tussilago farfara Extracts against Culex pipiens pallens and Tetranychus urticae)

  • 박성호;오현우;권혜리;서미자;유용만;윤영남
    • 농업과학연구
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    • 제41권3호
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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.