• 제목/요약/키워드: HPLC Chromatography

검색결과 2,219건 처리시간 0.03초

유황오리로부터 항종양 활성 성분의 분리 및 정제 (Isolation and Purification of Antitumor Substance from the Sulfur Fed Duck)

  • 윤원호;황진용;김창한
    • 한국축산식품학회지
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    • 제24권3호
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    • pp.293-297
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    • 2004
  • 본 연구에서는 유황오리로부터 항종양 효과를 나타내는 물질을 용매 추출법과 각종 크로마토그래피를 사용하여 분리 및 정제하였다. 유황오리로부터 정제한 활성 물질의 각종 종양세포에 대한 항암 효과를 측정한 결과 MTT assay는 100 $\mu\textrm{g}$/mL의 농도에서 HEp-2는 56%, KB는 58%의 세포 증식 억제 효과가 나타났다. 정상 세포주인 MDBK는 28%의 세포증식억제 효과가 나타나 정상 세포에 대한 세포 독성은 나타나지 않았다. Clonogenic assay는 200 $\mu\textrm{g}$/mL의 농도에서 HEp-2는 26%, KB는 28%의 생존율을 나타내었다. 따라서 유황오리로부터 정제한 활성 물질은 후두암 세포주인 HEp-2와 구강암 세포주인 KB에서만 특이적으로 세포 증식 억제 효과를 나타내었다.

조구등이 뇌허혈을 유발시킨 백서에서의 뇌신경보호효과 (The Protective Effect of Uncariae Ramulus Et Uncus Extract on Middle Cerebral Artery Occlusion(MACO))

  • 강상렬;강승준;금현수;전연이;이은주;박치상;박창국;허진화;양재하
    • 대한한의학회지
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    • 제23권1호
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    • pp.1-10
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    • 2002
  • Objectives: Uncariae Ramulus Et Uncus extract is clinically used in Korea to treat ischemic cerebral damage. The present study was undertaken to study the neuroprotective effect of Uncariae Ramulus Et Uncus extract in middle cerebral artery occlusion (MCAO) rats. Methods: Changes of extracellular levels of dopamine, DOPAC, HVA. and HIAA in striatum were collected at 20 minutes interval by in vivo microdialysis and then analyzed by HPLC (high performance liquid chromatography) in rats subjected to permanent focal cerebral ischemia induced by 2 hours of MCAO. Uncariae Ramulus Et Uncus extract was orally administrated before MCAO. Different animals were used for measurement of cerebral infarction volume induced by 24 hours of MCAO with TIC staining and image analysis. Results: Extracellular levels of dopamine decreased after treatment with Uncariae Ramulus Et Uncus extract, while extracellular levels of DOPAC and HVA significantly increased. Cerebral infarction volume also significantly decreased after treatment with Uncariae Ramulus Et Uncus extract. Conclusions: These results provided evidence that Uncariae Ramulus Et Uncus extract can produce a neuroprotective effect on cerebral ischemia by regulating extracellular excitatory neurotransmitters.

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Purification and Characteristics of Chitosanase from Bacillus sp. HW-002

  • Lee , Hyean-Woo;Choi, Jong-Whan;Han, Dong-Pyou;Park, Myoung-Jin;Lee, No-Woon;Yi, Dong-Heui
    • Journal of Microbiology and Biotechnology
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    • 제6권1호
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    • pp.19-25
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    • 1996
  • Chitosanase from Bacillus sp. HW-002 was purified with CM-cellulose column chromatography, and HPLC with DEAE- TSK gel and YMC-pack Diol 120. The purified enzyme appeared as a single band on SDS-polyacrylamide gel. The molecular weight of the enzyme was estimated to be about 46 kDa on SDS-polyacrylamide gel, and was estimated to be about 23 kDa by GFC. The optimal pH of chitosanolytic activity was about pH 5.5-6.0, and the purified enzyme was most stable at pH 5.0. The optimal temperature of chitosanolytic activity was $65^{\circ}C$ and the enzyme was stable at $45^{\circ}C$ for 1 h. Chitosan was the most favorable substrate among various $\beta$-glucan. UVmax of the purified enzyme was 195 nmand was not noted around 280 nm. The main product of enzyme reaction with chitosan was chitobiose.

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Gamnamoside, a Phenylpropanoid Glycoside from Persimmon Leaves (Diospyros kaki) with an Inhibitory Effect against an Alcohol Metabolizing Enzyme

  • Varughese, Titto;Rahaman, Mozahidur;Kim, No-Soo;Cho, Soon-Chang;Moon, Surk-Sik
    • Bulletin of the Korean Chemical Society
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    • 제30권5호
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    • pp.1035-1038
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    • 2009
  • Phytochemical investigation of the methanolic extract of Diospyros kaki leaves led to the isolation of osmanthuside H (1) and a new phenol glycoside, named gamnamoside [4-(3-hydroxypropyl)-2-methoxyphenol $\beta$-D-apiofuranosyl( 1 $\rightarrow$ 6)$\beta$-D-glucopyranoside] (2) along with (-) catechin (3) through a series of reversed phase column chromatography and preparative C18 HPLC. The structures of the isolates were determined by spectroscopic methods including IR, UV, HRTOFMS, and 2D NMR. Compounds 1, 2, and 3, showed good inhibitory activities ($IC_{50}$) of 175.4, 94.4, and 126.6 ${\mu}g/mL$ respectively, whereas a reversible ADH inhibitor, 4-methylpyrazole, showed the $IC_{50}$ of 326.6 ${\mu}g/mL$ against alcohol dehydrogenase (ADH).

Inhibitory Effect of Korean Fermented Vegetable (Kimchi) on the Growth and Aflatoxin Production of Aspergillus parasicitus-Part 1.

  • Kim, Jong-Gyu;Lee, Kyung-Min
    • 한국환경보건학회지
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    • 제33권3호
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    • pp.190-194
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    • 2007
  • Aflatoxin $B_1$ is a mycotoxin produced by Aspergillus flavus and A. parasiticus and is a human carcinogen. This study was performed to investigate reduction of growth and aflatoxin production of A. parasiticus by kimchi. A. parasiticus was grown in a modified APT broth with the juice of kimchi (at a concentration of 7%) at $28^{\circ}C$ for 9 days. Aflatoxin $B_1$ was determined by use of high performance liquid chromatography (HPLC). The addition of the juice of kimchi significantly reduced mycelial growth and aflatoxin production during the incubation period (p<0.05). Reduction of mycelial growth of A. parasiticus as the result of addition of the juice of kimchi was observed to range between 64.8 to 83.4% while reduction of aflatoxin production ranged from 62.2 to 73.0%. This study indicates that kimchi could be an effective inhibitor of aflatoxin production although mycelial growth may be permitted. More research is needed to study the inhibitory effects of the metabolites of kimchi.

Solid-Phase Extraction of Caffeine and Catechin Compounds from Green Tea by Caffeine Molecular Imprinted Polymer

  • Jin, Yinzhe;Row, Kyung-Ho
    • Bulletin of the Korean Chemical Society
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    • 제28권2호
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    • pp.276-280
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    • 2007
  • In this work, caffeine and some catechin compounds + C, EC, EGC, and EGCG were extracted from green tea by using molecular imprinted polymers (MIP) as sorbent materials in a solid-phase extraction (SPE) process known as MISPE (molecular imprinted solid-phase extraction). For synthesis of MIP, caffeine was employed as the template, MAA as the monomer, EGDMA as the crosslinker, and AIBN as the initiator. A solution of caffeine (0.2 mg/mL in methanol) was utilized in the solid extraction cartridges following loading, washing, and elution procedures with acetonitrile, methanol, and methanol-acetic acid (90/10, %v/v) as the solvents, respectively. This solid-phase extraction protocol was applied for the extraction of caffeine and some catechin compounds from green tea. A comparison was made between the results obtained with the MIP cartridges and a traditional C18 reversed-phase cartridge. It was thereupon found that the recovery of caffeine by the MIPbased sorbent used in this work was almost two and four times greater than that by a commercially available C18 material. A quantitative analysis was conducted by high performance liquid chromatography (HPLC) using a C18 column (5 μm, 250 × 4.6 mm) with methanol/water (40/60, %v/v) as the mobile phase at a flow rate of 0.5 mL/min.

호장근(Reynoutria elliptica)으로부터 항균활성 물질의 분리 및 구조결정 (Isolation and identification of antifungal compounds from Reynoutria elliptica)

  • 황주태;박영식;김영신;김진철;임치환
    • 농업과학연구
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    • 제39권4호
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    • pp.583-589
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    • 2012
  • In the continued research on natural fungicides for the control of plant diseases by using plant-derived products, we found that Reynoutria elliptica. had a strong fungicidal activity against several plant pathogens. R. elliptica (3.2 kg) were extracted with 80% aq. MeOH and the concentrated extracted was partitioned with n-hexane, EtOAc, n-BuOH and $H_2O$ successively. The four layers were tested their disease control efficacies against 4 plant disease such as rise blast (RCB), tomato grey mold (TGM), tomato late blight (TLB), and barly powdery mildew (BPM). The n-hexane fraction was highly active showing over 95% control against TLB and BPM. and the EtOAc fraction was highly active showing over 95% control against RCB, TLB, and BPM. By using silica gel chromatography, MPLC, and HPLC, three compounds that were expected to have antifungal activity were isolated. Their chemical structures were elucidated as physcion, emodin, and emodie-8-O-glucoside by EI-MS and NMR spectroscopic analyses.

지렁이(Lumbricus terrestris)로부터 Thiol-Specific Antioxidant protein(TSA)의 분리 및 정제에 관한 연구 (Purification and Characterization of TSA from Lumbricus terrestris)

  • 곽병구;김일한;차미경
    • 자연과학논문집
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    • 제14권2호
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    • pp.55-65
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    • 2004
  • 지렁이(Earthworm, Lumbricus terrestris)로부터 thiol-specific antioxidant activity(TSA)를 나타내는 향산화 단백을 분리정제 하였다. 이 향산화 단백은 환원제로 thiol성분에 의해 유지되는 비효소적 금속 촉매 산화계 (MCO, $Fe^{3+}$, DTT 또는 2-mercatoethanol ; Thiol- MCO system)에 의하여 Glutamine Synthetase의 불활성을 억제하지만 아스코르브산과 같은 nonthiol 환원제를 가진 효소적 금속 촉매 산화계 (MCO, $Fe^{3+}$, ascorbate ; nonthiol- MCO system)에서는 Glutamine Synthetase의 불활성을 방어하지 못하였다. 정제된 지렁이 TSA 단백질은 SAS-PAGE에 의해 51-kDa임을 밝혔고, 기존에 알려진 TSA protein과 분자량이 다른 TSA Family로써 활성 산소종에 의한 산화적 손상을 방어하는 향산화 효소로써의 중요한 생화학적 역할을 수행함을 제시하였다.

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1,2,4-트리메틸벤젠에 피폭된 노동자의 뇨 중 3,4-디메틸히푸르 산과 크레아티닌의 정량 (Determination of 3,4-Dimethylhippuric Acid and Creatinine from the Urine of Workers Exposed to 1,2,4-Trimethylbenzene)

  • 임정미;장재연;이용문;강종성
    • 약학회지
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    • 제41권6호
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    • pp.692-697
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    • 1997
  • Creatine and 3,4-Dimethylhippuric acid (3,4-DMHA), a glycine conjugate of 1,2,4-trimethyl-benzene (1,2,4-TMB) were determined in the urine of workers exposed to 1,2,4-TMB vapor. The best condition for the simultaneous determination of 3,4-DMHA and creatine by high performance liquid chromatography was obtained by reverse phase $C_{18}$ column (4.6${\times}$150mm, 5${\mu}m$) as stationary phase and 20% acetonitrile in 20mM phosphate buffer (pH 3.0) containing 4mM sodium octylsulfate(SOS)as mobile phase. The recovery of 3,4-DMHA spiked to blank urine in the range of 1~5${\mu}g$/ml was about 96%. The concentration of urinary 3,4-DMHA of workers had a positive correlation with the environmental level of 1,2,4-TMB (r=0.866). The data suggest that urinary 3,4-DMHA concentration is a useful biological index for 1,2,4-TMB exposure.

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뱀장어 유어(Anguilla anguilla)에 있어서 말라카이트 그린의 잔류성

  • Bergwerff, Aldert A.;Kuiper, Raoul V.;Scherpenisse, Peter
    • 한국양식
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    • 제19권1호
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    • pp.43-51
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    • 2007
  • 말라카이트그린(MG) 및 L-말라카이트그린(LMG)의 소멸은 뱀장어 Anguilla anguilla 유생에서 100일정도 걸린다. 실뱀장어(평균체중: 4.1 g)가 $23.0-26.5^{\circ}C$의 다양한 수온범위에 24시간동안 아주 낮은 농도(0.1 mg/l MG)에 약욕을 실시하였다. 처리한 후 실뱀장어는 말라카이트그린이 없는 깨끗한 물이 들어 있는 수조로 옮겼다. 뱀장어가 수용된 수조에는 순환여과 된 사육수를 공급하였다. 일정한 시간 간격으로 수집된 10마리의 뱀장어 및 사육수의 샘플은 high-performance liquid chromatography (HPLC)으로 분석하였다. 말라카이트그린의 가장 높은 농도는 $435{\pm}59\;{\mu}g/kg(mean{\pm}S.D.)$로서 처리시작 후 6시간에서 나타났다. L-말라카이트그린(LMG) 대사물의 농도는 비교적 높게 나타났으며 ($>100\;{\mu}g/kg$), 처리시작 후 6시간 및 처리종료 후 1500시 사이에서 안정적으로 나타났고 최대농도는 $831{\pm}231\;{\mu}g/kg$이었다. 비록 뱀장어는 체중이 증가하지는 않았지만 노출 후 24시간 후에도 L-말라카이트그린(LMG)은 여전히 전어체 kg당 $15{\pm}12\;{\mu}g/kg$ 함량이 존재하였다. 말라카이트 그린은 처리 이후 1920 및 2400시에서는 발견되지 않았다. 생물적 여과사육수와 순환여과시스템에서는 말라카이트그린이 발견되지 않았다. 본 연구에서는 부가적으로 빠르게 성장하여 11개월 된 뱀장어 두 마리를 샘플하여 분석에 사용하였다. 두 마리의 뱀장어는 실뱀장어기(0.3g)일 때 $0.15\;mg\;l^{-1}$ 농도의 말라카이트그린이 처리되었었다. 뱀장어의 근육조직에서는 지질 크로마토그래피-질량 분광광도계분석에 의해 말라카이트그린과 L-말라카이트그린이 측정 가능한 양으로 함유되지 않음을 확인할 수 있었다($<\;0.2\;{\mu}g\;kg^{-1}$).

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