• 제목/요약/키워드: purification method

검색결과 853건 처리시간 0.02초

Novel enzymatic elimination method for the chromatographic purification of ginsenoside Rb3 in an isomeric mixture

  • Cui, Chang-Hao;Fu, Yaoyao;Jeon, Byeong-Min;Kim, Sun-Chang;Im, Wan-Taek
    • Journal of Ginseng Research
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    • 제44권6호
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    • pp.784-789
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    • 2020
  • Background: The separation of isomeric compounds from a mixture is a recurring problem in chemistry and phytochemistry research. The purification of pharmacologically active ginsenoside Rb3 from ginseng extracts is limited by the co-existence of its isomer Rb2. The aim of the present study was to develop an enzymatic elimination-combined purification method to obtain pure Rb3 from a mixture of isomers. Methods: To isolate Rb3 from the isomeric mixture, a simple enzymatic selective elimination method was used. A ginsenoside-transforming glycoside hydrolase (Bgp2) was employed to selectively hydrolyze Rb2 into ginsenoside Rd. Ginsenoside Rb3 was then efficiently separated from the mixture using a traditional chromatographic method. Results: Chromatographic purification of Rb3 was achieved using this novel enzymatic elimination-combined method, with 58.6-times higher yield and 13.1% less time than those of the traditional chromatographic method, with a lower minimum column length for purification. The novelty of this study was the use of a recombinant glycosidase for the selective elimination of the isomer. The isolated ginsenoside Rb3 can be used in further pharmaceutical studies. Conclusions: Herein, we demonstrated a novel enzymatic elimination-combined purification method for the chromatographic purification of ginsenoside Rb3. This method can also be applied to purify other isomeric glycoconjugates in mixtures.

Edwardsiella tarda에 대한 계란난황항체의 분리와 정제 (Isolation and purification of chicken egg yolk immunoglobulin against Edwardsiella tarda)

  • 김영대;오명주;정태성;정성주
    • 한국어병학회지
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    • 제17권1호
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    • pp.11-20
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    • 2004
  • Edwardsiella tarda로 면역한 닭의 난황항체 (IgY) 정제 방법을 비교하였다. Anti-E. tarda IgY의 정제는 PEG법, chloroform-PEG법, ammonium sulfate법과 정제 kit를 사용한 4가지 다른 방법으로 실시하였다. 정제된 IgY는 64 kDa의 heavy chain과 27 kDa의 light chain을 나타내었다. E. tarda로 면역된 IgY는 면역되지 않은 대조 IgY 보다 높은 ELISA가와 응집항체가를 나타내었으며, 정제된 IgY는 western blotting에서 anti-E. tarda 토끼혈청과 유사한 E.tarda 단백질을 인식하였다. PEG법과 ammonium sulfate법에 의해 정제된 IgY는 응집항체가가 1:512, chloroform-PEG법과 정제 kit에 의해 정제된 IgY는 1:128을 나타내었으며, PEG법이 IgY를 정제하기 위한 가장 빠른 방법이었다. 이 연구의 결과로 PEG법이 IgY의 생물학적 활성을 유지함과 더불어 신속하고 효과적인 정제방법임을 알 수 있었다.

저콜레스테롤 난황 제조시 생성되는 부산물로부터 콜레스테롤의 분리 정제 (Separation and Purification of Cholesterol from By-product of Low Cholesterol Egg Yolk)

  • 유익종;조혜연;박우문;전기홍;최성유
    • 한국축산식품학회지
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    • 제20권1호
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    • pp.36-43
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    • 2000
  • $\beta$-cyclodextrin adsorption and saponification methods were applied to isolate and purify cholesterol from the by-product of the low-cholesterol egg yolk product. They by-product was prepared from processing low-cholesterol egg yolk followed by extracting with chloroform to remove $\beta$-cyclodextrin and concentrated to 3,069 mg% cholesterol. When $\beta$-cyclodextrin method between two purification methods was applied, 50% ethanol as a solvent showed higher cholesterol concentration of 5.82% rather than the other solvents. Repeated purification of 3 times could not improve the cholesterol concentration significantly(p<0.05). In case of purification using saponification method, hexane as a solvent for extraction of unsaponificated materials was more efficient to increase cholesterol concentration than chloroform and ether. 60 times(v/w) saponification solution (95% ethanol:33% KOH = 94:6) of sample weight was most effective to increase the cholesterol concentration of 35.7%. Repeated purification process by saponification method could increase cholesterol concentration to 95.7% by 4 times repetition.

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Novel Purification Method of Kv 4.2 Potassium Channel from Rat Brain Membrane

  • Park, Sung-Soo
    • 대한의생명과학회지
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    • 제18권2호
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    • pp.96-103
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    • 2012
  • Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.

냉각체 회전법에 의한 고순도 알루미늄 및 규소의 응고 및 정련에 관한 연구 (A Study on the Solidification and Purification of High Purity Aluminium and Silicon by Stirring Method)

  • 김욱;이종기;백홍구;윤우영
    • 한국주조공학회지
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    • 제11권4호
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    • pp.303-313
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    • 1991
  • The Purification mechanism of high purity aluminum was studied through the variation of stirring speed and coolant flow rate in the stirring method. In the stirring method the degree of purification was changed as the following factors;the variation of diffusion boundary layer thickness the variation of growth rate and the solute concentration of the residual melt. The concentration of Fe and Si was decreased as the stirring speed and the radial distance increased. In a high stirring speed of 2000rpm with unidirectional stirring mode, the uniformity of solutes was obtained. On the other hand, the purification of Si was done by the combinations of stirring method, fractional melting and acid leaching. In the case of Si purification, the centrifugal force developed in the melt acted as the significant purification factor. It was possible to obtain the purified 3N grade Si crystal after the complete elimination of residual aluminum by fractional melting and acid leaching.

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Purification of the Candida utilis Extracellular Invertase using Affinity Chromatography

  • Ginalska, G.;Belcarz, A.;Lobarzewski, J.;Leonowicz, A.;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • 제30권3호
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    • pp.12-17
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    • 2002
  • The extracellular invertase (EC 3.2.1.26) (Candida utilis) preparation was obtained from the liquid medium after desalting and freeze drying. This prepared enzyme was used for the comparative purification on 4 activated matrices by liquid column affinity chromatography method. In this method there were used controlled porous glass (CPG) silanized covalently activated by keratin, silanized silica gel and silica gel covalently covered by keratin. It was found that the invertase purification process was better using both CPG matrices (silanized CPG and keratin activated CPG) than these with two silica gel supports. Also the elution coefficient of the invertase from the two CPG columns was about 93 to 94%. Two silica gel supports found to be superior in terms of purification efficiency. The invertase purification process was confirmed by PAGE electrophoresis.

식물세포 배양으로부터 Peroxidase 대량 정제를 위한 전처리 공정 개발 (A New large-scale Pre-purification for Peroxidase from Plant Cell Cultures)

  • 표상현
    • KSBB Journal
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    • 제15권4호
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    • pp.342-345
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    • 2000
  • 식물세포배양으로부터 peroxidase를 대량 생산하기 위한 분리/정제 공정 으로서 세포를 파쇄하고 크로마토그래피 전처리로서 활성백토를 적용하였다. 활성백토는 미세한 세 포 조각 뿐만 아니라 여려가지 불순불을 선택적으로 흡착 하는 성질을 나타내었으며, 이를 통하여 효과적으로 정제 공정을 진행할 수 있었다 흡착을 통한 전처리 후 한외여 과장치를 이용하여 농축을 실시 하였으며, DEAE-Sepharose F FF를 이용한 크로마토그래피를 통해 정제가 이루어졌다. 활성을 나타내는 용액을 탈염, 농축 후 동결 건조하였다. 이 공정은 상업화에 필요한 대량 정제를 가능하게 하였으며 수율과 정제비용 측면에서 상당히 경제적인 공정임을 입증하였다, 활성백토를 이용한 흡착공정은 다른 효소 및 단백질의 경제적인 대량정제에 적용될 수 있으리라 기대된다.

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Ginsenoside의 새로운 분리.정제 방법 (New Efficient Method for Isolation and Purification of Ginsenosides)

  • 김세원;황석연
    • Journal of Ginseng Research
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    • 제22권4호
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    • pp.284-288
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    • 1998
  • This study was carried out to establish a new efficient method for isolation and purification of ginsenosides. Silica gel column chromatography, having been used for the isolation of ginsenosides, is advantageous to obtain a large amount of ginsenosides. However, it has a disadvantage to isolate ginsenosides to their highest purity. In addition, normal-or reverse-phase HPLC method thus far reported is confined to quantitative analysis. Especially, it has not been possible to isolate racemic 20(S)- and 20(R)-ginsenoside Rg2. In this experiment, isolation and purification of ginsenosides were accomplished by Diaion HP-20 adsorption chromatography, silica gel column chromatography, recrystalization and Prep. HPLC with or without Prep. TLC. From this study, we could establish a new efficient method for isolation and purification of 9 major and/or minor ginsenosides.

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TiO2 광촉매 나노구조에 따른 광유체 미세반응기 정수 효과 (Effects of the TiO2 nanostructures for water purification in optofluidic microreactor)

  • 권현아;김혜정
    • 한국가시화정보학회지
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    • 제20권3호
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    • pp.42-48
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    • 2022
  • The shortage of available freshwater is a major global issue worldwide and an increasing demand for clean water requires efficient water purification strategies. Here we describe a method to drastically increase the efficiency of a microreactor for photocatalytic water purification. To find out how the shape of the catalyst affects water purification, nanostructured catalysts of different structures, such as dense film, nanorod, and nanohelix, are prepared and their water purification characteristics are analyzed. Compared to the flat catalyst, the nanostructured catalyst showed a distinct ability in its pollutant degradation, but the detailed structural variation does not significantly affect the water purification. To further increase efficiency, we apply a micromixer to nanorod-based microreactor, which allows even enhanced mass transfer. This enables the solution of the water purification problem and greatly contributes to the industries where the efficiency of photocatalytic activity has attracted extensive interest.

Development of High Performance Liquid Chromatography for Paclitaxel Purification from Plant Cell Cultures

  • Kim, Jin-Hyun;Choi, Hyung-Kyoon;Hong, Seung-Suh;Lee, Hyun-Soo
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.204-210
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    • 2001
  • Paclitaxel can be produced in high yield and with a high degree of purify from plant cell cultures of Taxus chinensis. The complete purification method was systematically established and described. This method was an efficient procedure for the purification of paclitaxel from crude paclitaxel, consisting or reverse-phase chromatography, followed by a normal-phase chromatography. The two-stage HPLC purification scheme serves as an effective and economical approach for resolving paclitaxel from complex mixtures of taxoids, with high purify (>99%) and low impurities (<0.1%). The process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced. The process has been optimized to minimize solvent usage, complexity, and operating costs.

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