• 제목/요약/키워드: u-HPLC

검색결과 123건 처리시간 0.03초

황해의 용존 유기물 특성 및 아미노산 조성 (Amino Acid Composition and characteristic of dissolved organic Compounds in the Yellow Sea)

  • 박용철;윤철호
    • 한국해양학회지
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    • 제29권2호
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    • pp.171-182
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    • 1994
  • 황해 해역에서 해양 유기물의 생지화학적 특성을 파악하기 위하여 용존 자유 아미 노산 조성 및 추출된 유기물의 용존 가수분해 아미노산(DFAA) 조성 그리고 D/L alanine racemic ratio를 분석하였다. 용존 자유 아미노산의 농도는 평균 0.06 uM에서 0.26 uM의 범위를 보음, 우점하는 아미노산으로는 aspartate, glutamate, glycine, serine,alanin으로 나타났다. 각 기단별 물리·화학적 특성에 따라 분류한 아미노산 group 중 해수 중에 우점하는 것은 대부분 극성이 큰 hydrophilic group으로 나타났 다. 용존 가수분해 아미노산의 농도는 생거대유기물질의 경우 평균 2.05 uM에서 6.19uM의 범위를 보였고, 지구거대유기물질의 경우 평균 8.13 uM에서 24.46uM의 범위 를 보였다. 우점하는 아미노산의 group은 자유 아미노산과 유사한 경향을 보였다. 이 러한 용존 가수분해 아미노산은 생거대유기물질보다는 지구거대유기물질에서 상대적으 로 높은 농도를 보였다. 수괴가 성충화된 정점의 용존 가수분해 아미노산 농도는 저층 에 비해 표층이 상대적으로 높은 농도를 보였다. Alanine의 D/L racemic ratio는 생거 대 유기물질의 경우 0.126에서 0.146의 범위를 보였고 지구거대유기물질의 경우 0.309 에서 0.386의 범위를 보여 지구거대유기물질이 생거대유기물질보다 상대적으로 존재 연령이 오래된 것으로 나타났다. 이러한 자료들은 수괴의 특성 뿐만 아니라 생물활성 과 밀접하게 연관된 용존 유기 화합물의 중요한 생지화학적 지표를 이용될 수 있을 것 으로 사료된다.

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LC-MS/MS를 이용한 Homocysteine 측정과 그 유용성 평가 (Evaluation of the Method for Total Homocysteine in Plasma Using LC-MS/MS)

  • 전선희;임미숙;정영순;송정한
    • 대한임상검사과학회지
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    • 제37권1호
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    • pp.22-26
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    • 2005
  • Total homocysteine is now considered a risk factor for cardiovascular diseases. I increased interest has led to a multitude of studies requiring the determination of total homocysteine in conjunction with other factor. There are various methods for measuring total homocysteine, including HPLC, FPIA, GC-MS and LC-MS/MS. The most recent method for measuring total homocysteine uses a deuterium-labelled internal standard and tandem mass spectrometry. This development requires no derivatization and therefore leads to an increase in sample throughput compared to other techniques. We have evaluated the method for homocysteine by the LC-MS/MS method, and the correlation between the FPIA method and the LC-MS/MS method. The standard curve (0, 5, 10, 20, 50, 100 uM) was linear over the range examined (up to 100 uM). The lower limit of quantification (CV < 10 %) was 0.5 uM/L and the lower limit of detection (S/N >3) was 0.1 uM/L. Intra-assay variation and inter-assay variation were both <6 %. The comparision study for homocysteine concentration showed good correlation (r=0.9684) between the FPIA method and LC-MS/MS methods. Our conclusion is that the method showed relatively good precision, and was rapid and accurate.

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Pretreatment and enzymatic saccharification process of rapeseed straw for production of bioethanol

  • Lee, Heon-Hak;Jeon, Min-Ki;Yoon, Min-Ho
    • 농업과학연구
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    • 제43권4호
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    • pp.641-649
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    • 2016
  • This study was conducted to evaluate the yield of bio-ethanol produced by separate hydrolysis and fermentation (SHF) with the pretreated rapeseed straw (RS) using crude enzyme of Cellulomonas flavigena and Saccharomyces cereviase. Crude enzyme of C. flavigena showed enzymatic activity of 14.02 U/mL for CMC 133.40 U/mL, for xylan 15.21 U/mL, for locust gum and 15.73 U/mL for rapeseed straw at pH 5.0 and $40^{\circ}C$, respectively. The hemicellulose contents of RS was estimated to compromise 36.62% of glucan, 43.20% of XMG (xylan + mannan + galactan), and 2.73% of arabinan by HPLC analysis. The recovering ratio of rapeseed straw were investigated to remain only glucan 75.2% after 1% $H_2SO_4$ pretreatment, glucan 45.44% and XMG 32.13% after NaOH, glucan 44.75% and XMG 5.47% after $NH_4OH$, and glucan 41.29% and XMG 41.04% after hot water. Glucan in the pretreatments of RS was saccharified to glucose of 45.42 - 64.81% by crude enzyme of C. flavigena while XMG was made into to xylose + mannose + galactose of 58.46 - 78.59%. Moreover, about 52.88 - 58.06 % of bio-ethanol were obtained from four kinds of saccharified solutions by SHF using S. cerevisiae. Furthermore, NaOH pretreatment was determined to show the highest mass balance, in which 21.22 g of bio-ethanol was produced from 100 g of RS. Conclusively, the utilization of NaOH pretreatment and crude enzyme of Cellulomonas flavigena was estimated to be the best efficient saccharification process for the production of bio-ethanol with rapeseed straw by SHF.

식물 유래 조효소에 의한 감소 Liquiritin의 Liquiritigenin으로의 변환 (Biotransformation of Liquiritin in Glycyrrhiza uralensis Fisch Extract into Liquiritigenin by Plant Crude Enzymes)

  • 박민주;나인수;민진우;김세영;양덕춘
    • 한국약용작물학회지
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    • 제16권2호
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    • pp.74-78
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    • 2008
  • 감초의 에틸아세테이트 분획 중 flavonoid 지표물질인 liquiritin이 들어있는 분획 (GUE6)에 살구 및 복숭아 종자로부터 얻은 조효소액 (PDE, PAE, PPE)을 법제 처리하였다. 각 조효소액에서 ${\beta}-glucosidase$ 활성도는 아몬드 (P. dulcis) 259.6 U/g 살구 (P. persica), 복숭아 (P. persica) 조효소액의 ${\beta}-glucosidase$ 활성도가 가장 높게 관찰되었다. PDE, PAE, PPE를 이용한 발효 법제 후 liquiritigenin의 함량 비교 결과, 효소중의 ${\beta}-glucosidase$에 의해 liquiritin이 대사되어 항산화, 향군, 세포 독성 억제, 항치매, 항피부암 등 많은 약리효능을 가진 활성 물질인 liquiritigenin이 생산됨이 확인되었으며, 세효수 모두 liquiritin에 1.2배의 효소 처리 시 가장 대사가 활발한 것으로 나타나 변환을 위한 최적 농도로 결정되었다 세효소 중 특히 PPE 처리 시 liquiritin이 모두 liquiritigenin으로 변환됨으로써 liquiritin의 변환에 복숭아 종자 유래 효소가 가장 효율적인 것으로 밝혀졌다.

작약의 혈소판 응집억제작용에 관한 연구 (Study on Inhibition of Platelet Aggregation of Bioactive constituents from Paeonia lactiflora)

  • 서범석;박관혁;손동주;박영현;장성근
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2003년도 춘계학술발표논문집
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    • pp.320-321
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    • 2003
  • 최근 천연물을 중심으로 한 학문이 발전하면서 천연물이 가지는 생리활성 물질에 대한 관심이 중대되고 있다. 또한 인공합성품의 일부가 안정성의 문제가 제기되면서 천연물의 이용분야는 더욱 확대되고 있다. 급속한 경제발전과 생활수준의 향상으로 식생활을 포함한 생활방식의 다양화로 인하여 과거 감염 위주의 질병이 감소하고 선진국형의 만성퇴행성 질환이 증가하는 추세이다. 혈소판은 혈전증(thrombosis)과 지혈증(haemstasis)에서 중요한 역할을 담당하고 있는 인자로서 현관 내 병적 이상으로 인한 과도한 혈전의 생성은 뇌 -심혈관계 질환의 중요한 유발인자로 작용하므로 뇌ㆍ심혈관계 질환이나 항고지혈증을 연구하는데 유용하게 이용되고 있다. 본 연구에서는 토끼 혈액에서 분리한 세정 혈소판 부유액을 이용하여 작약 MeOH 추출물에서 분리한 천연생리활성물질들을 대상으로 혈소판 응집억제활성에 대하여 연구하였다. 작약 MeOH 추출물의 혈소판 응집억제활성 측정에서 강한 혈소판 응집억제활성 작용을 보였다. 따라서 작약 MeOH 추출물을 크로마토그라피법을 이용하여 분리하였고, NMR을 이용한 분광학적 방법으로 지금까지의 분리한 자료와 비교ㆍ분석하였다. Monoterpene glycoside 계열의 성분들인 15개의 compound와 다수의 fraction들을 HPLC를 이용하여 분리하였으며, collagen으로 유도된 혈소판 응집억제활성측정 방법에서 뛰어난 응집억제활성을 보였다. 표준물질을 이용한 HPLC 분석과 ¹H-NMR 관련 자료의 검색을 통하여 최종적으로 compound 1b가 benzoyloxy-paeoniflorin(2.6%). compound 1d가 paeond(1.3%), compound 2c가 albiflorin(3.2%), compound 2e가 paeoniflorin(33.6%)임을 확인할 수 있었다. Compound 3a의 분석결과 benzoyloxypaeoniflorin과 구조적 유사성은 있으나 동일한 구조식으로 확인할 수 없었다. 그러나 collagen에서 응집억제활성이 90% 이상으로 뛰어난 활성을 나타내므로 benzoyloxypaeoniflorin과 유사한 구조에서 benzoyl group이 다른 작용기로 치환되었거나 R₁ group이 다른 작용기로 치환된 형태로 추측하였다. Benzoyloxypaeoniflorin은 collagen>thrombin>U46616>A.A(arachidonic acid)>PAF의 순으로 활성을 보였다. 이는 paeoniflorin의 glycoside 5-carbon위치에 위치한 OH기 대신에 benzoyl기로 치환된 benzoyl 기가 혈소판 억제 산물로 작용한 것으로 추측했다. Paeoniflorin.은 U46619>thrombin>collagen>A.A>PAF순으로 억제를 보였다. Paeoniflorin이 collagen보다 thrombin에서 강한 억제를 보이는 것으로 Ca/sup 2+/ chelate를 형성함으로 인해 calciu 대사를 저해하는 것으로 추축했다. Compound 3a는 U46619>collagen>A.A>thrombin>PAF순으로 억제율을 보이므로 이 화합물은 paeoniflorin의 benzoyl기에 OH기가 다른 치환기로 바뀌거나 paeoniflorin의 glycoside 5-carbon 위치의 OH기 대신에 다른 작용기로 치환된 것으로 추정하였다. 이러한 결과로 작약의 주성분인 paeoniflorin과 유사한 구조를 가진 다른 monoterpene glycoside 계열의 화합물들과 비교 분석하고 구조를 화인하고 이들 성분이 어떻게 혈소판 응집억제활성에 작용하는지를 연구하였다.

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Purification and Characterization of a Chitinase from Cytophaga sp. HJ Isolated from Sea Sand

  • Lee, Dong-Mi;Noh, Hee-Jung;Lee, Kang-Man
    • Journal of Microbiology and Biotechnology
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    • 제9권6호
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    • pp.839-846
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    • 1999
  • An extracellular chitinase-producing bacterial strain induced by colloidal chitin was isolated from sea sand and was identified to be a member of the genus Cytophaga. The chitinase was purified successively by 30-60% ammonium sulfate fractionation, and DEAE-Bio gel A column, Octyl-Sepharose CL-4B column, and DEAE-Bio gel A column chromatographies. The enzyme had a molecular mass of 59.75 kDa, and the amino terminal amino acid sequence was ATPNAPVISW MPTDXXLQNXS. The enzyme acted better on colloidal chitin as a substrate than on chitosan. For colloidal chitin and chitosan (Degree of Acetylation, 15-25%), $K_{cat}$ values were 0.60U/mg and 0.08U/mg, respectively. HPLC analysis of the enzymatic reaction products showed that the chitinase produced mostly N-acetyl-D-glucosarnine and di-N-acetylchitobiose. The optimum temperature and pH for the enzyme were $50^{\circ}C$ and 4.0, respectively. N-Bromosuccinimide and $Hg^{2+}$ inhibited the chitinase activity as much as 90%, and $Sb^{3+}$, diethylpyrocarbonate, and $Ag^{+}$ inhibited it by 50-70%.

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Gentisyl Alcohol Inhibits Apoptosis by Suppressing Caspase Activity Induced by Etoposide

  • KIM JINHEE;KIM DONGHYUN;KIM MEEREE;KWON HOJEONG;OH TAEKWANG;LEE CHOONGHWAN
    • Journal of Microbiology and Biotechnology
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    • 제15권3호
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    • pp.532-536
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    • 2005
  • In the course of our screening for small molecules to inhibit apoptosis of U937 human leukemia cells induced by etoposide ($10\;{\mu}g/ml$), Penicillium sp. F020150 with potent inhibitory activity was selected. The active compound was purified from ethyl acetate extract of the microorganism by Sephadex LH-20 column chromatography and HPLC, and was identified as gentisyl alcohol (2,5-dihydroxybenzyl alcohol) by spectroscopic methods. The compound inhibited caspase-3 induction with $IC_{50}$ value of $3.0\;{\mu}g/ml$ after 8 h of etoposide treatment. The expression levels of caspase-3 and PARP were dose-dependently inhibited by the compound, suggesting that gentisyl alcohol inhibits etoposide-induced apoptosis via downregulation of caspases.

Antioxidant and Antidiabetic Activities of Ulmus davidiana Extracts

  • Guo, Jia;Wang, Myeong-Hyeon
    • Food Science and Biotechnology
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    • 제16권1호
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    • pp.55-61
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    • 2007
  • The antioxidant activities of water, ethanol, methanol, and chloroform extracts of Ulmus davidiana were evaluated using various antioxidant assays: DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical scavenging; hydroxyl radical scavenging; lipid peroxidation scavenging; and reducing power assays. All extracts, except the chloroform extract, demonstrated strong antioxidant activity in all assays. The chloroform extract had the highest hydroxyl radical scavenging activity, and its activity was equivalent to $\alpha$-tocopherol at a concentration of 0.5 mg/mL. Additionally, the antidiabetic activity of their extracts was also evaluated using a rat intestinal $\alpha$-glucosidase inhibition assay. Among all extracts investigated, the methanol extract had the highest $\alpha$-glucosidase inhibitory activity, although its activity was less than acarbose at $0.5\;{\mu}g/mL$. This result suggested that U. davidiana extracts may have antidiabetic activity. Total phenolic compounds and flavonoids were also measured. Phenolic compounds such as tannic acid, p-coumatric acid, and kaempferol were detected by high-performance liquid chromatography (HPLC). These results suggest that U. davidiana extracts may be useful as a potential source of antioxidant and antidiabetic materials.

가미온담탕(加味溫膽湯)이 스트레스성 뇌신경전달물질(腦神經傳達物質) 변화(變化)에 미치는 영향(影響) (Effect of Gami-Ondamtang(GO) on brain neuronal transmitters in immobilized rats)

  • 강탁림
    • 대한예방한의학회지
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    • 제5권2호
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    • pp.114-121
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    • 2001
  • To elucidate the preventive effect of oriental medicine Gami-Ondamtang(GO) on stress, we investigated the physiological change of rats which were applied immobilization stress. For immobilization stress, rats were placed in restrainer for 12 hours a day for 3 days. During application of stress, body weight of rats was measured. After sacrifice, 8 organs were taken for measurement of organ weight. Brain was sectioned into 4 parts that are Frontal Cortex, Corpus Striatum, Hypothalamus and Hippocampus. Each part was homogenated and its catecholamine and serotonin contents were measured with HPLC. In our study, stress mainly induced increase of concentration of neurotransmitters in brain without other significant physical change of rats. GO inhibited stress induced changes of neurotransmitter content in brain.

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Phenolic Acid Changes in Mycelia of Sclerotium rolfsii After Garlic and Onion Supplementation in a Broth Medium

  • Pandey, M.K.;Singh, D.P.;Singh, U.P.
    • Mycobiology
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    • 제33권3호
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    • pp.137-141
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    • 2005
  • High performance liquid chromatographic (HPLC) analysis of mycelia of Sclerotium rolfsii grown in broth medium supplemented with garlic (Allium sativum) and onion (Allium cepa) was carried out to estimate qualitative and quantitative changes in phenolic acids. Several phenolic acids, such as gallic, chlorogenic; ferulic, o-coumaric and cinnamic acids were detected in varied amounts in mycelia grown on such media as compared to control. Phenolic acids represents a wide range of secondary metabolite found in the cells of plants and microbes including fungi. The growth characters of S. rolfsii in various supplements also varied from thin and transparent to thick and opaque.