• 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.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.4 no.2
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• pp.189-197
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• 1994

Analytic methods for Cr(VI) level in industrial hygienic field were suggested by the National Institute for Occupational Safety and Health(NIOSH method 7600, 7604). There were growing needs for measurement of Cr(III) and Cr(VI) levels simultaneously. Two analytical methods were suggested to determine Cr(III) and Cr(VI) levels simultaneously. The one is method by using reversed phase high peformance liquid chromatography(HPLC) and the other is by using ion exchange HPLC. The purpose of this work was to evaluate the usefulness of these two analytic methods. For the difference of ionic charges of Cr(III)-ethylendiamine tetraacetic acid(EDTA) chelate and $CrO_4{^-2}$, we could detect them simultaneously by ion exchange HPLC. Also, we attempted to determine the levels of Cr(III) and Cr(VI) chelated with sodium diethyldithiocarbamate(NaDDTC) by using reversed phase HPLC. The confirmation of Cr(III) and Cr(VI) were checked by fraction collector and nameless atomic absorption spectrometer. The optimal conditions for the formation of Cr(III)-EDTA chelate were two hours incubation period with pH 5. Cr(III)-EDTA and Cr(VI) in EDTA solution were successfully separated by anion exchange column using $Na_2CO_3/NaOH$ mixture as mobile phase. Peaks of Cr(III)-EDTA and Cr(VI) in EDTA were identified at 5 minutes and 7 minutes of retention time respectively by the ion exchange HPLC. The formation of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were twelve hours incubation period. Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were separated by reversed phase column using methanol and water mixture as mobile phase. Peaks of Cr(VI)NaDDTC and Cr(III)-NaDDTC chelates were identified at 13 minutes and 26 minutes of retention time respectively by the reversed phase HPLC. Due to reduction of Cr(VI) to Cr(III), it seems to be not suitable for simultaneous determination of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates by reversed phase HPLS. Simultaneos determination of Cr(III) and Cr(VI) by ion exchange HPLC was more accurate and simple method.

• Korean Chemical Engineering Research
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• v.50 no.4
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• pp.659-665
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• 2012

IgY (Immunoglobulin Yolk) in egg yolk corresponds to IgG (Immunoglobulin G) in animal serum and plays an important role as immunological proteins in intestines. Carrageenan and Arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE (Diethylaminoethyl) Sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtration High Performance Liquid Chromatography) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatography) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at 0.5 M NaCl and pH=5.

• KSBB Journal
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• v.15 no.1
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• pp.76-79
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• 2000

Complex, ill-defined mixtures of natural origin are often used as nutrients in the production of biological products through microbial fermentation. Product yields are affected by variation in these natural products. Yeast extract is a typical example of these natural products. Since it is a mixture of amino acids, peptides and nucleic acids, its composition is not well characterized. In this study, we investigated the properties of thiamine hydrochloride, riboflavin and pyridoxine hydrochlride in yeast extract by using a gel filtration chromatography, ion exchange chromatography and high performance liquid chromatography. Yeast extract solution was fractionated by gel filtration chromatography and ion exchange chromatography, and then, each fraction was analyzed by using a high performance liquid chromatography.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.81-85
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• 2002

A marine bacterium producing superoxide dismutase, strain number B446, was screened with nitrite quantitation method using hydroxy amine and photochemically generated superoxide ion, and the superoxide dismutase was purified through 35-75% ammonium sulfate precipitation, DEAE-Sephadex A-25 ion exchange chromatography, Sephadex G-200 gel filtration chromatography, and High-Q anion exchange chromatography to a yield of 6% and purification fold of 32.3.

• KSBB Journal
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• v.23 no.1
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• pp.31-36
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• 2008

The use of L-carbohydrates and their corresponding nucleosides in medicinal application has greatly increased. For example L-ribose has been much in demand as the starting material for curing hepatitis B. High performance liquid chromatography (HPLC) method was studied for the analysis of ribose and arabinose fractions from ion exchange chromatography (IEC). Dowex Monosphere 99 Ca/320 resin was packed in IEC to separate ribose and arabinose under various operating conditions. $NH_{2}$ and sugar HPLC columns were then used to analyze the fractions from the IEC column. Pulse input method (PIM) was also used to measure adsorption isotherms of ribose and arabinose in the Dowex column and HPLC columns. Experimental results and simulations by ASPEN chromatography were compared with fair agreement.

• Korean Chemical Engineering Research
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• v.52 no.1
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• pp.98-105
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• 2014

Lysozyme amounts to 0.3% in egg white and functions as an agent of cell lysis and activator of tissue reconstruction. Ion exchange chromatography is the most useful method of separation among affinity chromatography, ion exchange chromatography, and ultra-filtration. The aim of present study is to find the optimum pH and temperature for the separation of lysozyme in egg white within cation exchange gel filled glass column. And we compared results of experiments with those of simulations. Phosphate buffer was used, and pH and temperature were varied as 5~7 and $25{\sim}40^{\circ}C$ respectively. RP-HPLC was the tool for the retention time identification and quantitative analysis of lysozyme. OriginPro 8 measured the peak area of lysozyme chromatogram and quantified the eluted lysozyme. Largest amount of lysozyme was separated under the conditions of pH 5 and T $25^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.22 no.1
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• pp.52-58
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• 1994

The induced antibiotic peptides were isolatde from the white-spotted folwer chafer, Protaetia brevitarsis by injection of E. coli suspension to the larvae of the insect. The antibacterial activity of the peptides were assayed by the plate growth ingibition method, and were purified by ion-exchange chromatography, reversed-phase HPLC, ion-exchange HPLC and SDS-PAGE etc. The peptides were estimated as 9 kDa in molecular weight and named Protaecin I and Protaecin II, respectively. Protaecin I and II have strong antibacterial activities against Gram-positivie and/or Gram-negative bacteria, and they are stable in the heat treatment and in the range of pH 2-12.

• The Korean Journal of Physiology
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• v.24 no.2
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• pp.363-375
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• 1990

The present investigation was performed to purify bombesin-like immunoreactivity (BBS-LI) from the skin of frogs, B. orientalis inhabiting Korea. For extraction of BBS-LI, the fresh skin of 360 g from frogs was immersed in 1,800 ml of 100% methanol and then kept at $4^{\circ}C$ for 5 days. BBS-LI was partially purified by liquid chromatography using an alkaline alumina column followed by a Sephadex G-10 column. BBS-LI was further purified by using sequential HPLC of reversed phase C18 preparation, gel permeation, SP-ion exchange and reversed phase C18 analysis. BBS-LI in fractions of each step was monitored by radioimmunoassay for which bombesin antiserum with a titer of 1 : 188,800 was raised in a guinea pig. Eventually, two different BBS-LI were successfully purified and each BBS-LI showed the following character. 1) BBS-LI was well separated into two peaks in SP-ion exchange HPLC. One (BBS-LI-K1) bound to the column while the other (BBS-LI-K2) did not. 2) BBS-LI-K1, 73.8% of total BBS-LI, was not differentiated from synthetic bombesin in reversed phase C18 analytical and gel permeation HPLC. 3) BBS-LI-K2, 26.2% of total BBS-LI, eluted later than synthetic bombesin in reversed phase C18 analytical HPLC, but it eluted with a retention time identical to that of synthetic bombesin in gel permeation HPLC. 4) The two forms of BBS-LI and synthetic bombesin identically stimulated gastrin release and pancreatic exocrine secretion including volume, protein output and amylase output in anesthetized rats. It is concluded from the above results that the skin of B. orientalis contains two different forms of BBS-LI which are very identical to bombesin immunologically and biologically. In comparison with synthetic bombesin containing 14 amino acid residues, the major form shows quite similar pattern in all HPLC used in the present study, but the minor form exhibits quite different pattern in SP-ion exchange and reversed phase C18 analytical HPLG.

• Journal of Pharmaceutical Investigation
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• v.27 no.4
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• pp.313-321
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• 1997

Ion exchange resin complexes of famotidine have been prepared by the reaction of famotidine solution with activated ion exchange resins. Complex formation efficiency between famotidine and ion exchange resin was about $80{\sim}90%$ in average, calculated by HPLC determination. Drug release characteristics from the resin complexes were evaluated by the modified percolation method. Famotidine release was dependent on the type of ion exchange resins. In the case of weakly acidic resin complexes, the cumulative released amount of famotidine was more than 90% for 1hr in pH 1.2 buffer solution. However, in the case of strongly acidic resin complexes, it was less than 5% for 3hr in the same medium. Strongly acidic resins revealed some advantages over weakly, acidic resins for overcoming instability of famotidine in gastric juice. In addition, strongly acidic resin complexes showed controlled release of famotidine in pH 6.8 buffer solution, showing the result of about 60 to 70% of drug release for 5hr. After oral administrations of famotidine-resin complexes to rats as dose of 40 mg equivalent/kg, the pharmacokinetic parameters of famotidine were obtained by model independent analysis and compared with those of famotidine solution or suspension. $C_{max}$ of famotidine-resin complex was lower than that of famotidine solution or suspension. MRT, MAT, and MDT of the complexes were greater than those of famotidine solution or suspension. From these results, it was expected that famotidine was released slowly from the complexes and absorbed continuously into systemic circulation. It was recognized that drug release from the complexes was the rate-limiting step in drug absorption, since there were close correlations between in vitro drug release and in vivo pharmacokinetic parameters.