• Title/Summary/Keyword: HPLC Chromatography

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Isolation of Herbicidal Compounds from Pulsatilla koreana Roots (백두옹(Pulsatilia koreana Nakai) 뿌리로부터 제초활성물질의 분리)

  • 정형진
    • Korean Journal of Plant Resources
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    • v.9 no.1
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    • pp.47-54
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    • 1996
  • To search herbicidal compounds in Pulsatilla koreana Nakai, methanol extract of P. koreana roots was purified by sequences of XAD-7 column chromatography, silica gel adsorption column chromatography, silica gel flash column chromatography, preparative layer chromatography and preparative high performance liquid chromatography(Prep, HPLC).The final Prep. HPLC gave two herbicidally-active fractions. These fractions treatment at 100ppm inhibited the root length of Echinochloa crus-galli seedlings by 48% and 60% as compared with the control, respectively. Components in the two active fractions were analyzed by GC-MS Spectrometry. These compounds, which were isolated from P. koreana roots, were identified as several fatty esters, hydrocarbons, squalene, evidonol, and a diazepin analogue.

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Mutation Analysis in β2-Adrenergic Receptor Gene by Denaturing High Performance Liquid Chromatography (DHPLC) (DHPLC를 이용한 β2-교감신경수용체 유전자에서의 돌연변이 분석)

  • Park, Sang-Bum;Oh, Chung-Hun;Kim, Jong-Wan;Jang, Won-Cheoul
    • Analytical Science and Technology
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    • v.15 no.3
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    • pp.190-195
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    • 2002
  • We analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed-phase high performance liquid chromatography (IP-RP-HPLC). We extracted genomic DNA from 50 asthma patients, amplified DNA using PCR, and analysed PCR product by DHPLC. As a result, we obtained that mutation frequency was 15 (30%) among 50 cases. Consequently DHPLC mutation detection was confirmed that the result of direct sequencing was coincide exactly.

Recent Developments in High-performance Liquid Chromatography of Lipids

  • Christie, William W.
    • Journal of the Korean Applied Science and Technology
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    • v.10 no.1
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    • pp.1-8
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    • 1993
  • The possibilities for HPLC analysis of lipids have been revolutionised by the availability of evaporative light-scattering detectors, with which the response is independent of the nature of the mobile phase and does not depend On the presence of specific chromophores in the lipids. It was thus possible to develop an HPLC procedure, involving ternary gradient elution, for separating all the lipid classes in animal tissues in a single step. Although reversed-phase HPLC has been widely used for the analysis of molecular species of lipids, sliver ion chromatography can be a valuable alternative. For example, a stable silver ion column for HPLC was developed which permitted resolution of molecular species of triacylglycerols, even from such complex samples as fish oils, again With light-scattering detection and gradient elution. The capacity for HPLC resolution of diastereomeric diacyl-sn-glycerol derivatives, prepared from triacylglycerols. has lead to a new simple method for stereospecific analysis of the latter.

Study on the molecular weights of radioprotective ginseng proteins by HPLC method

  • Kim, Choon-Mi;Kim, Hyeon-Lim
    • Archives of Pharmacal Research
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    • v.9 no.1
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    • pp.5-9
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    • 1986
  • Partialy purified ginseng proteins were either treated with sodium dodecyl sulfate (SDS) and .betha.-mercaptoethanol to denature the proteins or not, and subjected to Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC) to compare the components of each fraction. Standard proteins of known molecular weights (MW) were also either treated with SDS and .betha.-mercaptoethanol or not, and subjected to HPLC to obtain regression lines for MW determination. From the retention times obtained from samples in eiether case by HPLC, the MW were estimated as following in SDS treated condition, Gl fraction showed three peaks each with MW of above 100, 000, 51, 000 and 19, 000. Gll showed one original peak with MW of 21, 000 and Gll, two peaks each with MW of 19, 000 and 14, 000. On the other hand, in non-SDS treated condition, GI fraction showed two peaks each with MW of above 200, 000 NA 52, 000. Gll showed one original peak with MW of 41, 000 and Gill, three peaks each with MW of 28, 000, 19, 000 and 14, 000.

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Large Scale Purification of KRF-001 on the Preparative HPLC (Preparative HPLC를 이용한 KRF-001의 대량분리정제)

  • 이항우;김무경정태숙복성해
    • KSBB Journal
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    • v.9 no.4
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    • pp.385-394
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    • 1994
  • Large scale purification to get antifungal antibiotic KRF-001 of 90% purity, was investigated using preparative HPLC. Crude KRF-001 was purified by XAD-7 adsorption chromatography, acid precipitation and microfiltration. Microfiltration was the most effective isolation method of crude KRF-001. The purification methods using C18 chromatography was convenient compared with the conventional methods. Delta PAK C18 column and Bonda PAK C18 column were adapted large scale purification of KRF-001. Gradient system of prep HPLC using Delta PAK C18 column was more effective. With these conditions, final recovery of KRF-001 yielded 77%.

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Determination of Monocrotophos Residues by HPLC

  • Lee, Young-Deuk;Kwon, Chan-Hyeok
    • Korean Journal of Environmental Agriculture
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    • v.23 no.4
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    • pp.245-250
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    • 2004
  • An analytical method was developed to determine monocrotophos residues in apple, citrus, and soil using high-performance liquid chromatography (HPLC) with ultraviolet absorption detection. Monocrotophos was extracted with acetone from apple, citrus and moist soil samples. The extract was concentrated, added with saline water, and subjected to n-hexane washing to remove nonpolar co-extractives. Dichloromethane partition was then followed to recover monocrotophos from the aqueous phase. Silica gel column chromatography was employed to further purify the extract prior to HPLC determination. Reverse-phase HPLC using an oct-adecylsilyl column was successfully applied to separate and quantitate the monocrotophos residue in sample extracts at the wavelength of 230 nm. Overall recoveries of monocrotophos from fortified samples averaged $95.3{\pm}2.1%$ (n=6), $970{\pm}0.7%$ (n=6), and $92.8{\pm}4.3%$ (n=12) for apple, citrus, and soil, respectively. The proposed method was quite reproducible and sensitive enough to replace the troublesome gas-liquid chromatographic analysis for monocrotophos residues.

Analytical method for analyzing formaldehyde using 2, 4-DNPH and gas chromatography/FID, NPD (2,4-DNPH와 가스크로마토그래프를 이용한 포름알데히드 분석방법)

  • Jeong, Jee Yeon;Park, Seung Hyun;Yi, Gwang Yong;Oh, Se Min
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.10 no.1
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    • pp.126-146
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    • 2000
  • To develop and evaluate formaldehyde measurement method using 2,4-dinitro-phenylhydrazine (2,4-DNPH) coated sampler and gas chromatography, laboratory test and field test were conducted. Results of this study are as follows. Limit of detection(LOD) of measurement methods, HPLC-UVD, GC-NPD and GC-FID, is $0.008{\mu}g/m{\ell}$ $0.060{\mu}g/m{\ell}$, $0.472{\mu}g/m{\ell}$ respectively. Coefficiency of measurement methods, HPLC-UVD, GC-NPD and GC-FID, is 0.008, 0.009, 0.020 respectively. Desorption efficiency of sep-pak xposure aldehyde sampler and sorbent sample tube is 1.05(range : 0.99 - 1.12), 1.02(range : 0.99 - 1.06) respectively. Samples of sorbent sample tube and sep-pak xposure aldehyde sampler turned out to be stored at refrigerator, according to storage test results. Measurement methods of HPLC-UVD, GC-NPD, GC-FID, according to results of precision for the combined sampling and analytical procedure, became acceptable to OSHA evaluation standard. Field test using exposure chamber met the NIOSH overall uncertainty recommendation(less than 25%). Overall uncertainty of Sepak-HPLC(UVD), Tube-GC(NPD), Tube-GC(FID) is 11.0% - 17.0%. Consequently gas chromatography(GC-NPD, GC-FID) and high performance liquid chromatography(EPA TO-11) using 2,4-DNPH coated sampler for formaldehyde measurement turned out to be suitable to measure personal formaldehyde exposure at workplaces.

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Isolation of 3,4-Dihydroxycinnamic Acid with Antimicrobial Activity from Bark of Aralia elata (두릅수피에서 항미생물 활성을 갖는 3,4-Dihydroxycinnamic Acid의 분리)

  • Ma, Seung-Jin;Kuk, Ju-Hee;Ko, Byoung-Seob;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.28 no.3
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    • pp.600-603
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    • 1996
  • The methanol extract of Aralia elata bark showed antimicrobial activities against bacteria, yeast and fungi. The active components were successively purified with solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, silica gel partition column chromatography and HPLC. The active substances were separated with HPLC where 1% acetic acid-MeOH (60 : 40, v/v) was used as mobile phase. The isolated active substance ($t_r$ 17.1 min) was identified as trans-3,4-dihydroxycinnamic acid by $MS,\;^1H-NMR\;and\;^13C-NMR$.

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Autotoxicity of alfalfa flower extract and its allelopathy to Echinochloa crus-galli (알팔파 꽃 추출물의 Autotoxicity와 돌피에 대한 Allelopathy)

  • Ill Min, Chung;Song Joong, Yun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.6
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    • pp.821-832
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    • 1997
  • The aim of this study was to separate or purify some bioactive compounds from flowers of alfalfa(Medicago sativa L.) and to test of the isolated compounds on alfalfa for their autotoxicity and on Echinochloa crus-galli for their allelopathy for seed germination and seedling weight. Using thin layer chromatography(TLC) of $CHCl_3$ extracts, the most inhibitory band to alfalfa seed germination was determined. Germination inhibition of this extract suggested a complex chemical interaction. Separation and purification of compounds with CHCl$_3$ extract of fresh alfalfa flowers were conducted by a silica gel TLC, and microcrystalline cellulose TLC(MCTLC), followed by droplet countercurrent chromatography(DCCC) bioassay. Preliminary identification was done by high perfomance liquid chromatography(HPLC) on the most inhibitory fractions in DCCC. Ferulic acid, caffeic acid, vanillic acid, rutin, narringin were identified in fraction 5 and ferulic acid, caffeic acid, vanillic acid, rutin, coumarin in fraction 6. The phytotoxicity of their individual compound was tested on alfalfa and Echinochloa crus-galli seed germination and seedling weight. Coumarin and ferulic acid showed the most inhibitory effect on alfalfa seed germination and Echinochloa crus-galli seedling fresh and dry weight. These compounds may be, at least in part, involved in autotoxicity and allelopathy.

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Brassinosteroid substances in immature oryza sativa seeds (벼종자의 brassinosteroid 활성물질)

  • Park, Keun-Hyung;Kim, Seon-Jae;Park, Jong-Dae;Lee, Lan-Sook;Hyun, Kyu-Hawn
    • Applied Biological Chemistry
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    • v.36 no.5
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    • pp.376-380
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    • 1993
  • To investigate the presence of the brassinosteroid substances in immature Oryza sativa L. cv Tongjinbyeo seeds, the methanol extract was purified by the sequential use of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography and charcoal adsorption chromatography. The activity of brassinosteroid was monitored by the rice inclination test and its presence could be confirmed in each purification step. The purified active components were separated by silica gel adsorption chromatography and Sephadex LH-20 chromatography. Brassinosteroid substances in separated active fractions were identified as castasterone, teasterone and 6-deoxocastasterone by HPLC. Our work is probably the first report of endogenous brassinosteroids in Oryza sativa seeds. The content of brassinosteroid in Oryza sativa seeds as converted into brassinolide was $0.5{\sim}1.5\;ng/g$ fresh weight.

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