• Title/Summary/Keyword: HPLC 분리

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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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A Study on the Separation of n-Hexane by Molecular Sieve 5A and the Purification for HPLC use (분자체 5A를 이용한 n-헥산의 분리와 HPLC급으로의 정제에 관한 연구)

  • Choi, Beom Suk;Kim, Young Man;Kim, Sun Tae
    • Analytical Science and Technology
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    • v.6 no.1
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    • pp.21-27
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    • 1993
  • Technical grade n-hexane whose purity is 54% has been purified for HPLC use. Methylcyclopentane, 2-methylpentane, and 3-methylpentane which are hardly isolated by fractional distillation were separated by the liquid-solid chromatography using molecular sieve 5A. UV and fluorescence impurities whose contents are critically regulated for HPLC solvent were removed by the adsorptive separation with alumina and silica gel. The present method also reduced the impurities of color(APHA), acidity, water, residue after evaporation, sulfur, and thiophene content, and the impurity contents were well within the specifications of HPLC solvent.

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Identification of Proteins in Egg White Using Ion Exchange Cartridge and RP-HPLC (이온교환 카트리지와 RP-HPLC를 이용한 난백 단백질의 확인)

  • Kim, Hyun Moon;Kim, Ah Reum;Lee, Chang Soo;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.713-717
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    • 2012
  • Approximately forty proteins in egg white have been widely studied for their functional properties. To develop a procedure of separation for pure and non-altered proteins from egg white, purification study was conducted to isolate lysozyme, ovotransferrin, and ovalbumin. Ion exchange cartridge can selectively separate proteins from egg white, and reversed-phase HPLC (RP-HPLC) could identify separated proteins. Proteins in egg white were purified by HI trap ion exchange cartridge SP and Q with buffers pH 8.0 and 5.2. C18 column (Phenomenex, USA) was used for RP-HPLC analysis and isocratic mobile phase was used with acetonitrile (ACN)/distilled water (DW)/trifluoroacetic acid (TFA) in the ratio of 50/50/0.1. Comparing the retention times of standards in RP-HPLC experiments showed that ovotransferrin, ovalbumin, and lysozyme in egg white were eluted successively in the RP-HPLC column after the pretreatment in SP and Q ion exchange cartridges.

Determination and Ethylene Oxide(EO) Separation of Polysorbates by RP-HPLC (역상 HPLC에 의한 polysorbates의 산화에틸렌(EO) 분리 및 정량분석)

  • Lee, Yong-Hwa
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.4
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    • pp.585-593
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    • 2012
  • The EO separation and quantitative determination of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80 was carried out by reversed phase HPLC. The water/acetonitrile was used for the mobile phase of gradient conditions. An YMC Pack Ph ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and Phenomenex C4 ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and the selected ELSD detector was applied. The analysis results of HPLC showed good linearity with correlation coefficient of $r^2$=0.997 in the rage of $180.2{\sim}980.5{\mu}g/mL$ and detection limit.

Purification of 4-Methylthio-3-butenyl isothiocyanate the Pungent Principle in Radish Roots by RP-HPLC (RP-HPLC법에 의한 무우의 매운 성분인 4-methylthio-3-butenyl isothiocyanate의 분리 및 정제)

  • Kim, Mee-Ree;Rhee, Hei-Soo
    • Korean journal of food and cookery science
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    • v.2 no.2
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    • pp.16-20
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    • 1986
  • The major pungent component from Korean radish roots was purified by reverse phase high performance liquid chromatography (RPHPLC), and characterized as 4-methylthio-3-butenyl isothiocyanate on the basis of the sensory test (pungency), UV spectrum and mass spectrum analysis. The purified isothiocyanate moved as a single peak(retention time, 5.2 min) in RP-HPLC analysis, and as a single spot(Rf, 0.9) in TLC analysis.

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Separation of ${\alpha}-,\;{\gamma}-,\;{\delta}-tocotrienol$ from latex (천연 latex로부터 ${\alpha}-,\;{\gamma}-,\;{\delta}-tocotrienol$의 분리)

  • Lee, Hyung-Ok
    • Applied Biological Chemistry
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    • v.36 no.1
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    • pp.29-32
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    • 1993
  • 3 $Tocotrienol-{\alpha}-,\;{\gamma}-\;and\;{\delta}-tocotrienol-from$from latex(Hevea Brasiliensis) were isolated and the oily tocotrienol concentrates obtained. To isolate tocotrienols, the fractionation by semipreparative HPLC of the unsaponifiable fraction in the raw lipid extract from latex was carried out. By this method, the total content of tocotrienols in latex was ca. 400 ppm, and the purities of ${\alpha}-,\;{\gamma}-\;and\;{\delta}-tocotrienol$ were 98.3, 99.3 and 96.2%, respectively.

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Isolation and Purification of Decursin and Decursinol Angelate in Angelica gigas Nakai (참당귀(Angelica gigas Nakai) 중 Decursin 및 Decursinol Angelate의 분리 및 정제)

  • Kim, Kang-Min;Jung, Jae-Yeon;Hwang, Sung-Woo;Kim, Myo-Jeong;Kang, Jae-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.653-656
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    • 2009
  • This paper is intended as an investigation of the method of extraction and the analysis by high-performance liquid chromatography mass spectroscopy of decursin and decursinol angelate in the dried root of Angelica gigas Nakai. The extracted decursin and decursinol angelate were the purity of >95% using 60% ethanol at $-20^{\circ}C$ for 12 hours by HPLC analysis. Decursin and decursinol angelate were efficiently isolated using recycling HPLC. The purity of isolated decursin and decursinol angelate was identified as 99.97 and 99.40% by HPLC analysis, respectively. The molecular weights of Decursin and decursinol angelate were also identified as m/z=329 ($[M+H]^+$) and m/z=351 ($[M+Na]^+$) by mass spectroscopy.

Bioactivity Analysis of Curcuminoids from Turmeric using On-line Screening HPLC-ABTS (On-line Screening HPLC-ABTS를 이용한 강황으로부터 Curcuminoids의 생물활성 분석)

  • Choi, Sun Do
    • Journal of Applied Biological Chemistry
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    • v.56 no.3
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    • pp.137-139
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    • 2013
  • Free radical scavengers in the bisdemethoxycurcumin (BDMC), demethoxycurcumin (DMC) and curcumin of turmeric (Curcuma longa) were screened, identified, quantified and isolation using coupled off-line-2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) and on-line screening high-performance liquid chromatography (HPLC)-ABTS assay. There was a very small margin of error between the off-line-ABTS method and the on-line screening HPLC-ABTS method.

Analyses of Additives Applied in a Polycarbonate (폴리카보네이트에 사용된 첨가제의 분석)

  • Kim, Seog-Jun
    • Analytical Science and Technology
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    • v.13 no.3
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    • pp.282-290
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    • 2000
  • In this study, polymer additives were extracted and separated by Soxhlet extraction method and the dissolution-precipitation method from a polycarbonate (optical grade) which completely absorbed UV light below 390 nm. Analytical techniques such as UV-Vis spectroscopy, FT-IR, and HPLC were applied to analyze additives in polycarbonate. Separated materials from the polycarbonate may be a complex mixture containing additives such as UV stabilizer, antioxidants (primary and secondary), monomers, and oligomers. Several compounds such as bisphenol A, Irganox 1010, and Cyasorb UV-5411 were identified by chromatograms and UV spectra obtained from RP HPLC analysis using Bondapak $C_{18}$ column, methanol mobile phase, and a photodiode array (PDA) detector. Also, the content of UV-5411 in the polycarbonate was about 0.12 wt% by a quantitative analysis through UV spectroscopy.

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Cobra (Ophiphagus hannah) 독으로부터 세포 독성 단백질의 분리

  • 김영식;이병무;안미영;박재갑;현진원;임경화;박호군
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.205-205
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    • 1994
  • 동물, 곤충, 해조류 등으로 부터 얻어진 독소는 해로운 것으로 알려져 있지만 의학적으로 유용한 화합물을 내포하고 있다. 본 연구는 이러한 점에 착안하여 독사, 또는 곤충의 독 30여종을 대상으로 위암 세포인 SNU-1에 대해서 MTT assay에 의한 세포 독성 시험을 실시하였다. 이 중 세포 독성 활성이 가장 놓은 코브라 계통인 Ophiphagus hannah의 독으로 부터 세포 독성 물질의 정제를 시도하였다. HPLC-GPC와 HPLC-lEX 또는 RP-HPLC를 이용하여 정제하였고 각 분획들을 암세포주에 대해서 시험하였다. TSK-2000SW로 부터 2개의 분획을 얻어 분획 I에서 $IC_{50}$/ 의 값이 0.5 - 3 $\mu\textrm{g}$/ml의 범위 내에서 확인되었다. 분자량이 작은 분획 II에서는 세포독성이 작게 나타났지만 농축하여 성분 분석을 시도하였다. 분획 I을 RP-HPLC를 이용하여 TFA와 acetonitrile의 linear gradient에 의해 더욱 분리를 하였고, phopholipase $A_2$ 활성의 가능성도 측정하였다. 분획 II는 Mono S 칼람을 이용하여 ammonium acetate농도에 따른 pH gradient에 의한 분리를 시도하여 순도를 SDS-PAGE에 의해서 확인하였다. 각각의 크로마토그리피로 부터 얻은 분획에 대해서 세포 독성을 실시중에 있으며 성질도 동정중에 있다.

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