• Title/Summary/Keyword: Elution chromatography

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Metabolite profiles of ginsenosides Rk1 and Rg5 in zebrafish using ultraperformance liquid chromatography/quadrupole-time-of-flight MS

  • Shen, Wenwen;Wei, Yingjie;Tang, Daoquan;Jia, Xiaobin;Chen, Bin
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.78-84
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    • 2017
  • Background: In the present study, metabolite profiles of ginsenosides Rk1 and Rg5 from red ginseng or red notoginseng in zebrafish were qualitatively analyzed with ultraperformance liquid chromatography/quadrupole-time-of-flight MS, and the possible metabolic were pathways proposed. Methods: After exposing to zebrafish for 24 h, we determined the metabolites of ginsenosides Rk1 and Rg5. The chromatography was accomplished on UPLC BEH C18 column using a binary gradient elution of 0.1% formic acetonitrile-0.1% formic acid water. The quasimolecular ions of compounds were analyzed in the negative mode. With reference to quasimolecular ions and MS2 spectra, by comparing with reference standards and matching the empirical molecular formula with that of known published compounds, and then the potential structures of metabolites of ginsenosides Rk1 and Rg5 were acquired. Results: Four and seven metabolites of ginsenoside Rk1 and ginsenoside Rg5, respectively, were identified in zebrafish. The mechanisms involved were further deduced to be desugarization, glucuronidation, sulfation, and dehydroxymethylation pathways. Dehydroxylation and loss of C-17 residue were also metabolic pathways of ginsenoside Rg5 in zebrafish. Conclusion: Loss of glucose at position C-3 and glucuronidation at position C-12 in zebrafish were regarded as the primary physiological processes of ginsenosides Rk1 and Rg5.

Antioxidative Activity of Hot Water and Ethanol Extracts of Lespedeza cuneata Seeds (야관문(Lespedeza cuneata) 종자의 열수 및 Ethanol 추출물의 항산화 효과)

  • Kim, Seon-Jae;Kim, Du-Woon
    • Food Science and Preservation
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    • v.14 no.3
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    • pp.332-335
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    • 2007
  • Hot water and 95%(v/v) ethanol extracts were prepared from dried Lespedeza cuneata seeds and antioxidant compounds were isolated by solvent fractionation, silica gel adslorption chromatography, Sephadex LH-20 column chromatography, and preparative HPLC. Antioxidant activity was measured using DPPH radical scavenging activity. The 80%(v/v)ethanol and ethylacetate fraction of Lespedeza cuneata seed extracts had stronger antioxidant effects than did the n-hexane fraction. The active antioxidant compounds obtained from hot water and 95%(v/v) ethanol extracts may be identical, based on analysis by Sephadex LH-20 column chromatography and preparative HPLC.

Rational and efficient approach to the preparation of the active fractions of Scutellaria baicalensis (황금(Scutellaria baicalensis) 유효분획물 제조의 합리적이고 효율적인 접근방법)

  • Kim, Doo-Young;Kim, Won Jun;Kim, Jung-Hee;Oh, Sei-Ryang;Ryu, Hyung Won
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.31-38
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    • 2019
  • Scutellaria baicalensis Georgi (Scutellariae Radix) has been widely used as a dietary ingredient and traditional herbal medicine such as diuretic, hyperlipidemia, antibacterial, anti-allergy, anti-inflammatory and anticancer properties. In this study, the isolation of biomarkers or bioactive compounds from complex S. baicalensis extracts represents an essential step for de novo identification and bioactivity assessment. The bioactive fraction consisted of eight compounds which was chromatographed on an analytical high performance liquid chromatography column using two different gradient runs. A simulative replacement of the analytical column with a medium pressure liquid chromatography and open column allowed the determination of gradient profile to allow sufficient separation in the preparative scale. From the optimized method, eight standard compounds have been identified in the fractions. In addition, MS, UV, HRMS detection was provided by ultraperformance liquid chromatographyequadrupole time-of-flight mass spectrometry (UPLC-QTof-MS) of all fractions. Therefore, this scale up procedure was successfully applied to a S. baicalensis extract.

Antioxidant activity of ethanol extract and methanol fractions via column chromatography from Psidium guajava Leaf (구아바 잎 추출물 및 컬럼크로마토그래피를 이용한 메탄올 분획물의 항산화 활성)

  • Byeoung-Kyu Choi
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.266-271
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    • 2023
  • The antioxidant capacity of the Psidium guajava leaf extracted with EtOH and their MeOH fractions using column chromatography were evaluated by 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging assays, total phenolic and flavonoid content, and Superoxide dismutase (SOD) assay. To determine its utility as a functional material, the crude extract was fractionated by flash column chromatography on ODS using a stepwise elution with combinations of MeOH/H2O and then all the fractions were also investigated. In the results of antioxidant activities, the 40% and 60% MeOH fractions show the meaningful values, and then the two fractions were selected to examine the isolation and identification of the major constituents via HPLC and nuclear magnetic resonance. Further purification led to isolation of two quercetin derivatives; quercitrin (1) and isoquercetin (2). Through SOD assay, some methanol fractions via column chromatography and isolated compounds showed improved antioxidant activities compared to the extract.

Induced Circular Dichroism Spectra and Chromatography by Interaction of Some Racemic Cobalt (III) Complexes and d-Tartrate$^2$- in Aqueous Solution (d-주석산이온과 라세미코발트(III) 착물과의 상호작용에 따른 유발원편광이색성 Spectra와 크로마토그래피)

  • Chang Eon Oh;Dae Ho Kang;Gab Choul Shin
    • Journal of the Korean Chemical Society
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    • v.25 no.5
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    • pp.306-310
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    • 1981
  • Induced circular dichroism spectra of racemic cobalt(III) complexes for $[Co(en)_3]^{3+},\;[Co(tn_)3]^{3+},\;cis-[Co(NH_3)(en)_2]^{3+},\;[Co({\beta}-ala)(en)2_]^{2+},\;[Co(gly)(en)_2]^{2+}\;and\;[Co(acac)(en)_2]^{2+}$ were measured when they were dissolved in aqueous d-tartrate2- solution at room temperature. Only a single negative CD spectrum was observed for all the complexes above in visible region(400∼500nm). It was interpreted that these CD bands were attributed to the difference in interaction between ${\Lambda}$-and ${\Delta}$-enantiomers with d-tartrate$^{2-}$. Namely, when d-tartrate$^{2-}$ was added to ${\Lambda}$-enantiomer and ${\Delta}$-enantiomer, it caused ${\Lambda}$-enantiomer to change greatly and ${\Delta}$-enantiomer to change only slightly; combined the results proved induced circular dichroism. The enantiomer for which the eluent has a stronger affinity should be eluted faster in ion-exchange column chromatography. It is possible to predict the elution order of chromatography from the sign of the induced CD if stronger interaction of chiral anion with the complex leads to greater change in the natural CD spectrum of the complex. The elution order was in complete agreement with the prediction from the sign of the induced CD spectrum for all the measured complexes.

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Optimization of solid phase extraction and simultaneous determination of trace anions in concentrated hydrofluoric acid by ion chromatography (불산 중 극미량 음이온 분석을 위한 고상 추출법 및 이온크로마토그래프를 이용한 동시분석법 확립)

  • Yoon, Suk-Hwan;Jo, Dong-ho;Kim, Hyun-Ji;Shin, Ho-Sang
    • Analytical Science and Technology
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    • v.29 no.5
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    • pp.219-224
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    • 2016
  • 불산 중 극미량 음이온의 고상추출과 이온크로마토그래프를 이용한 고감도 분석법이 개발되었다. 불산 중 불소이온이 고상에 의해 제거하였고 이어서 음이온 (F, CH3COO, Cl, Br, NO3, PO43−, SO42−)들이 이온크로마토그래프를 이용하여 연속적으로 분리하였다. 고상 추출법에 영향을 주는 각 인자들 (흡착제의 선택, 시료의 부피 및 pH, 용출 용액과 용출용액의 부피)을 결정하였으며 그 결과 흡착제로서 Oasis WAX 컬럼이 가장 우수하였고 1.0 mL의 시료부피, 용출용액으로 50 mM 초산암모늄염 5 mL가 분리능에서 가장 우수하였다. 개발한 방법에 의한 음이온 (Cl, Br, NO3, PO43−, SO42−)들의 방법검출한계는 25 % 불산용액 (w/w) 중에 0.04~0.30 µg/L의 범위를 보였고 정밀도는 20.0와 40.0 µg/L의 농도에서 5 % 이내를 보였다. 한 제조회사에 의한 25 % 불산 중 음이온의 4.2에서 47.5 µg/L의 범위로 모두 검출되었다. 이 방법은 시험절차가 간단하고, 재현성 및 감도가 좋아서 반도체회사에서 불산 중 음이온 불순물을 정도 관리하는데 매우 유용한 방법이 될 것으로 판단된다.

Optimization of Anion-exchange Chromatography for the Separation of Agarase from Culture Broth of Pseudoalteromonas sp. (Pseudoalteromonas sp. 배양액으로부터 아가레이즈 분리를 위한 음이온교환 크로마토그래피 최적화)

  • Kim, Yu-Na;Lee, Jae-Ran;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Hong, Soon-Kwang;Kim, Chang-Joon
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.840-845
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    • 2011
  • Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.

ELUTION OF RESIDUAL MONOMER ACCORDING TO VARIOUS LIGHT SOURCES AND CURING TIME ON THE POLYMERIZATION OF PHOTOACTIVATED PIT AND FISSURE SEALANTS (광중합 광원의 종류와 조사시간에 따른 치면열구전색제의 미반응 모노머 용출)

  • Oh, You-Hyang;Park, Yoon-Kyung;Lee, Nan-Young;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.3
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    • pp.421-430
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    • 2004
  • The purpose of this study was to measure and compare the amount of unreacted TEGDMA from pit and fissure sealants cured with three different light sources; conventional halogen light curing unit, plasma arc light curing unit and argon laser. The specimens were eluted in distilled water for different time intervals. The time-related release of TEGDMA were analyzed by reverse-phase high performance liquid chromatography(HPLC). The result of present study can be summarized as follows: 1. The time-related release of TEGDMA decreased with increasing curing time in conventional halogen light, however, that not statistically significant difference(p>0.05). 2. The elution from the specimens cured for 6 and 9 seconds with plasma arc light was similar results corresponding with the time-related TEGBMA release, and was significantly lower than that cured for 3 seconds(p<0.05). 3. The elution of TEGDMA from the specimens cured with argon laser was significantly higher than that cured with halogen and plasma arc light(p<0.05). 4. The elution of TEGDMA from under recommended time of three different light sources were showed to be no statistically significant difference(p>0.05). 5. In time-related release of TEGDMA from recommended time of each light sources, the results correspond to 40 seconds of halogen light and 6 seconds of plasma arc light were similar(p>0.05). 6. The elution of TEGDMA, from over recommended time of three different light sources were showed to be no statistically significant difference(p>0.05). In this study, I suggest that curing time of plasma arc light is 6 and/or 9 seconds in the field of clinical pediatric dentistry claiming its effectiveness in optimal polymerization and reduced chair time.

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Determination of Ginsenosides Content in Korean Ginseng Seeds and Roots by High Performance Liquid Chromatography

  • Hu, Jiang Ning;Lee, Jeung-Hee;Shin, Jung-Ah;Choi, Jae-Eul;Lee, Ki-Teak
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.430-433
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    • 2008
  • A high performance liquid chromatography (HPLC) method has been successfully developed to identify and quantify major ginsenosides in Korean ginseng seeds and roots. Using gradient elution of acetonitrile and water without buffer, the 6 major ginsenosides ($Rb_1,\;Rb_2$, Rc, Rd, Re, and $Rg_1$) were identified. Compared with ginseng roots, the amount of ginsenoside Re and Rd in ginseng seeds were significantly higher than those in ginseng roots (p<0.05). In ginseng seeds, the content of protopanaxtriol (PPT) was higher than that of protopanaxdiol (PPD) and the ratio of PPT and PPD was approximately 2.2 : 1. However, the content of PPT was lower than that of PPD in ginseng roots. It should be mentioned that both content of PPT and PPD in ginseng seeds were much higher than those in ginseng roots.

Screening of Antioxidative Compounds toward Human Low Density Lipoprotein (LDL) from Useful Plants (유용식물로부터 Human Low Density Lipoprotein(LDL)에 대한 항산화제의 탐색)

  • 임복규;류병호
    • The Korean Journal of Food And Nutrition
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    • v.17 no.2
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    • pp.138-146
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    • 2004
  • This study was designed to investigate antioxidative activity of substances isolated from 25 kinds of useful plants resources toward free radical and human low density lipoprotein(LDL). Methanol extracts of Oenothers odorate had the highest antioxidative activity similar with ${\alpha}$-tocopherol. Methanol extracts of Oenothers odorate was extracted again by the ethylacetate. The ethylacetate soluble acidic fraction obtained from methanol extract of Oenothers odorate showed highest activity toward human LDL. Each fraction was purified through Sepadex LH-20 chromatography by elution of chloroform-methanol mixture (90:10 v/v). Fraction, F-2 obtained from Oenothers odorate showed at highest levels of electron donating activity. Fraction, F-2 was identified as 3,4-dihydroxybenzoic acid and 3-hydroxycinnamic acid.