• Journal of Microbiology and Biotechnology
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• 제12권2호
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• pp.196-203
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• 2002

A novel process for urokinase purification was studied using p-aminobenzamidine as the ligand and sepharose 4B as the matrix. The adsorption, washing, and elution conditions were optimized by an unusual method. An adsorption buffer containing 2.5 M NaCl and $1\%$ Tween 80 facilitated the adsorption of urokinase on the affinity media and prevented contaminants from binding to the p-aminobenzamidine affinity gel. It was found that $5\%$ Tween 80 removed most of the contaminants from the affinity column. A 0.2 M glycine elution buffer containing 0.5 M NaCl (pH 3.0) was found to have a strong elution ability with a high recovery and purity of urokinase. A crude urokinase material of231 IU/mg protein from human urine was purified to 124,300 IU/mg protein with a purification factor of 538 and yield of $86.7\%$. As a result, a high purity urokinase was obtained with only a single affinity chromatography step. The purification process was successfully scaled-up to a 2-1 chromatography column. The resulting urokinase eluate could be directly lyophilized, thereby complying with Chinese pharmacopoeia (1995 version) standards.

• 분석과학
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• 제30권1호
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• pp.20-25
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• 2017

An analytical method for determining potential endocrine disruptors (bisphenol A, 2-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, pentachlorophenol, p-t-butylphenol, p-pentylphenol, p-hexylphenol, p-t-octylphenol, p-heptylphenol, nonylphenol) by solid-phase extraction (SPE) and High Perfomance Liquid Chromatography(HPLC) equipped with fluorescence and variable wavelength detector has been developed. The SPE process for sample concentration was performed on a commercially available Oasis HLB cartridge packed with polymeric sorbents. The effect of elution solvent and elution volume on the recoveries of the analytes were investigated with HPLC. Average recovery of >85% was achieved with 60mg sorbents using 5mL of methanol as elution solvent. Phenolic compounds in canned drinks, beverages and water samples were surveyed by this proposed method.

• Bulletin of the Korean Chemical Society
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• 제22권6호
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• pp.570-574
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• 2001

Magnesium isotope separation was investigated by chemical ion exchange with the 1-aza-12-crown-4 bonded Merrifield peptide resin using an elution chromatographic technique. The capacity of the novel azacrown ion exchanger was 1.0 meq/g dry resin. The heavier isotopes of magnesium were enriched in the resin phase, while the lighter isotopes were enriched in the solution phase. The single stage separation factor was determined according to the method of Glueckauf from the elution curve and isotopic assys. The separation factors of $^{24}Mg^{2+}$-$^{25}Mg^{2+}$, $^{24}Mg^{2+}$-$^{26}Mg^{2+}$, and $^{25}Mg^{2+}$-$^{26}Mg^{2+}$ were 1.008, 1.019, and 1.006, respectively.

• 한국동물위생학회지
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• 제36권4호
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• pp.273-281
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• 2013

This study investigated the effects of the alkaline detergent and acid rinse used for cleaning milking machines on the eight commercially available teatcup liner materials. The sample liners prepared for use in the clean-in-place process were analyzed by ultraviolet spectrophotometer, ion chromatography and liquid chromatography. Among the eight liner materials, the ultraviolet spectra of 3 sample liners were shown to have a similar peak shape after cleaning, but the ultraviolet spectra peak shape of 5 sample liners was noticeably changed. No products were detected by ion chromatography in any of the liner materials used in this study. When the liner materials were only treated with alkaline detergent, some additional peaks were observed using liquid chromatography which indicate the creation of molecular substance and elution from liner materials, however, these peaks disappeared when the liner materials were cleaned with the acid rinse. Therefore, we propose that an acid rinse should be applied, after cleaning the milking machine with the alkaline detergent.

• Mass Spectrometry Letters
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• 제12권4호
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• pp.186-191
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• 2021

A new method for chemical separation of light rare-earth elements (LREEs) using gas-pressurized extraction chromatography (GPEC) is described. GPEC is a microscale column chromatography system that features a constant flow of solvents, which is created by pressurized nitrogen gas. The separation column with a Teflon tubing was packed with LN resin. The proposed GPEC method facilitates production of lesser chemical wastes and faster separation owing to the use of low solvent volume compared to traditional column chromatography. We evaluated the separation of Ba, La, Ce, and Nd using various elution solvents. The column reproducibility of the proposed GPEC system ranged from 2.4% to 4.9% with RSDs of recoveries, and the column-to-column reproducibility ranged from 3.1% to 6.3% with RSDs of recoveries. The proposed technique is robust, and it can be useful for the fast separation of LREEs.

• KSBB Journal
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• 제14권3호
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• pp.371-376
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• 1999

제조용 액체 크로마토그래피를 전산모사하는 것은 최적의 분리와 조건을 위해 필수적이다. 제조용 액체 크로마토그래피에서의 단일 성분과 2성분 용출 피크의 거동을 설명하는 수학적 모델을 equilibrium-dispersive model을 이용해 계산을 수행하였다. 모델의 전산모사를 통해 주입되는 시료부피, 시료속의 용질의 농도, 유속, 컬럼길이가 용출되는 피크에 미치는 영향을 확인해 보았다. Equilibrium-dispersive model로 전산모사된 data를 이용하여 단일 성분 시료 주입에 대해 주입 농도와 유속에 대한 영향을 알아본 결과 주입 농도를 증가시킬수록 용출이 일어나는 시점이 앞당겨져 평균 체류 시간이 감소하고 날카로운 피크를 얻을 수 있다. 유속을 증가시키면 시료의 용출 시점이 앞당겨지고, 고정상과의 충분한 상호 작용을 일으키기 전에 용출되기 때문에 피크가 날카로와 진다. 2성분 시료 주입에 대해 분리도값들을 계산한 결과 두번째로 용출되는 시료의 농도를 증가시킬수록 이 시료성분이 더 빨리 용출되므로 분리도값이 감소함을 알 수 있다. 칼럼길이가 분리도에 미치는 영향을 보면 칼럼길이를 길게 할수록 시료성분이 고정상과의 완전한 상호작용으로 분리가 잘되고, 시료의 주입량을 증가시키면 두 성분간의 용출 피크의 간격이 좁아져 분리도 값이 감소함을 알 수 있다. 이 내용을 바탕으로 다성분 분리에 대한 simulation 연구로 확장할 수 있다.

• 환경위생공학
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• 제12권2호
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• pp.59-64
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• 1997

For purification of a 70kDa toxic protein of mosquitocidal delta-endotoxin from B. thuringiensis strain H9B, immuno-affinity chromatography was performed. After separation of 70kDa toxic proteins from the delta-endotoxin of the strain H9B on SDS-PAGE, the 70kDa toxic protein was subcutaneously injected into rabbit for making a polyclonal antibody. A anti-70kDa toxic protein was purified by a column chromatography packed with protein A-sepharose 4B gels. The 70kDa toxic protein from delta-endotoxin of the strain H9B was also purified by an immuno-affinity chromatography packed with CNBr-activated sepharose 4B gels conjugated anti-70kDa toxic protein after elution with 1/10M citric acid-1/5M Na$_{2}$HPO$_{4}$ buffer(pH3.2) containing 0.5M NaCl. The 70kDa toxic protein was purified through only one step-separation system, was demonstrated by SDS-PAGE and immunoblot.

• KSBB Journal
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• 제13권2호
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• pp.125-132
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• 1998

Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70${\mu}g/cm^2$ determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20 ${\mu}g/cm^2$ for flow rates above 6 mL/min.

• 한국식품과학회지
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• 제20권6호
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• pp.856-861
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• 1988

본 실험은 Staphylococcus aureus의 한 균주로부터 A와 C, 두 종류의 toxin이 동시에 생성될 때 이들의 동시분리를 위하여 각종 분리방법을 연구하였다. 혼합된 toxin A와 C는 CM-column chromatography를 이용하여 pH-gradient법으로 용출했을 때 2개의 분획이 나타났으나 서로 완전히 분리되지 않아 다량의 서로 다른 toxin이 함유되어 있었고 Sephadex G-75, Sephacryl S-300, 그리고 ultro gel을 사용한 gel filtration에서는 하나의 분획을 나타내 상호분리가 불가능 하였으며 정제도와 분리도에서 가장 뛰어난 gel column을 이용한 FPLC도 toxin A와 C를 상호분리할 수 없었다. 그러나 CM-column을 이용한 FPLC에서는 enterotoxin A는 pH 6.8에서 그리고 enterotoxin C는 pH 8.6에서 각각 분리되었으며, immunodiffusion test 결과 enterotoxin A의 분획에서는 toxin C가 전혀 검출되지 않았고 enterotoxin C의 분획에서도 toxin A가 검출되지 않았다. 용출방법에 있어서는 CM-column을 이용한 FPLC에서 pH-stepwise법이나 pH-gradient법으로 enterotoxin A와 C type을 쉽게 동시에 분리할 수 있었다.

• 분석과학
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• 제7권3호
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• pp.379-386
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• 1994

Ni(II), Cu(II), Co(III) 및 Cr(III) 이온과 2-hydroxy-arylazopyrazolone 유도체들 사이에 형성하는 킬레이트들에 대한 역상 액체 크로마토그래피에서의 용리 메카니즘을 열역학적인 접근방법으로 고찰하였다. van't Hoff plot에 의해 온도와 용량인자(k')와의 상관관계를 조사한 결과 온도와 용량인자와의 관계는 양호한 직선성을 나타냈으며, 온도 변화와 무관한 동일한 메카니즘에 의해 분리됨을 알 수 있었다. van't Hoff plot으로부터 엔탈피$({\Delta}H)^{\circ}$, 엔트로피$({\Delta}S)^{\circ}$를 구하고, 엔탈피와 용량인자와의 상관관계를 조사한 결과 [pm(2-OH)(5-Cl)PaPz](r=0.787)을 제외한 대부분이 킬레이트들은 비교적 좋은 직선성(r=0.980~0.999)을 보였고, 보정온도(${\beta}$) 역시 일정한 값을 나타내었다. 이것은 금속-2-hydroxy-arylazopyrazolone 킬레이트와 정지상($C_{18}$)과의 상호작용이 등평형 거동을 하고 있음을 의미한다. 또한 엔탈피와 용량인자와의 상관관계로부터 구한 보정온도는 374.63~806.9k 범위의 값을 나타냈고, 이들 결과로부터 역상 액체 크로마토그래피에서 금속 킬레이트들의 머무름 메카니즘은 소수성 효과에 기인하고 있음을 확인할 수 있었다.