• 한국식품위생안전성학회지
• /
• 제32권5호
• /
• pp.404-410
• /
• 2017

HPLC-size-exclusion chromatography에 의해 가공식품에서 알긴산프로필렌글리콜의 함량을 분석하는 방법이 개발되었다. 알긴산프로필렌글리콜을 분석하기 위해 GF-7M HQ column과 LT-ELSD detector가 선정되었다. 알긴산프로필렌글리콜 분석을 위한 전처리 조건으로는 $20^{\circ}C$에서 150 rpm으로 3시간 동안 추출하는 방법이 선정되었다. 알긴산프로필렌글리콜을 5 농도(300, 500, 700, 1,000, and 1,500 mg/kg) 범위에서 검량선을 작성한 결과 직선성($R^2$)은 0.9873으로 측정되었다. HPLC system에 의한 알긴산프로필렌글리콜 분석시 검출한계(LOD) 및 정량한계(LOQ)는 각각 171.43 mg/kg 및 519.50 mg/kg이었다. Size-exclusion chromatography에 의해 얻은 회수율 및 변동 계수(coefficient of variation)는 각각 86.1~110.4% 및 4.1~13.5%이었다. 본 연구에서 개발된 HPLC-size-exclusion chromatography system을 적용하여 134 품목의 가공식품에서 알긴산프로필렌글리콜 함량을 분석하였다. 이 결과들은 이 방법이 가공식품에서 알긴산프로필렌글리콜 함량을 분석하는데 적용할 수 있는 방법이라는 것을 나타낸다.

• Journal of Microbiology and Biotechnology
• /
• 제19권4호
• /
• pp.416-423
• /
• 2009

Hepatitis B core antigen(HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus(HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the "hold-up" period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.

• 분석과학
• /
• 제8권4호
• /
• pp.405-410
• /
• 1995

A study on elemental speciation of trace metals in lake water (Lake Biwa in Japan) has been carried out by a size exclusion chromatography (SEC) / inductively coupled plasma mass spectrometry (ICP-MS) system. Before analysis, the water sample was preconcentrated with a ultrafiltration technique, where the large molecules with molecular weight larger than 10,000 were concentrated. Then the preconcentrated water samples (500-1000 fold) were analyzed by a SEC/ICP-MS system. Most trace metals were found at the UV absorption peaks corresponding to the molecular weights of ca. 300,000 and 10,000-50,000, where trace metals were on-line detected by ICP-MS. The results suggest that many of trace metals exist as the large organic molecules-metal complexes in natural water.

• 한국생명과학회:학술대회논문집
• /
• 한국생명과학회 2002년도 제37회 국제학술심포지움 및 추계학술대회
• /
• pp.9-17
• /
• 2002

Renaturation of Lysozyme by size exclusion chromatography(SEC) to improve yield as well as the initial and final protein concentration has been studied in detail, Although urea decreases the rate of proteins refolding, it can suppress protein aggregation to sustain pathway of correct refolding at high protein concentration, and there existed an optimum urea concentration in renaturation buffer. Lysozyme was successfully refolded from initial protein concentration of up to 100mg/m1 by SEC, the yield was more than 40%. And the refolding of Interferon-${\gamma}$ was further investigated.

• Molecular & Cellular Toxicology
• /
• 제1권2호
• /
• pp.87-91
• /
• 2005

Although 2D-PAGE has been widely used as the primary method for protein separation, difficulties in displaying proteins with an extreme values of isoelectric paint (pI), molecular size and hydrophobicity limit the technique. In addition, time consuming steps involving protein transfer and extraction from the gel-pieces can result in sample loss. Here, we describe a novel protein separation technique with capillary size-exclusion chromatography (CSEC) for rapid protein identification from human plasma. The method includes protein fractionation along with molecular size followed by in-solution tryptic digestion and peptide analysis through reversed phase liquid chromatography (RPLC) coupled to nanoflow electrospray-tandem mass spectrometry (ESI-MS/MS). Tryptic peptides are applied an a $100\;{\mu}m\;i.d.{\times}10mm$ length pre-column and then separated on a $75\;{\mu}m{\times}200mm$ analytical column at -100 nL/min flaw rate. Proteins were identified over the wide ranges of pI (3.7-12.3) when this technique was applied to the analysis of $1-2\;{\mu}L$ of human plasma. This gel-free system provides fast fractionation and may be considered a complementary technique to SDS-PAGE in proteomics.

• 한국식품과학회지
• /
• 제22권1호
• /
• pp.33-37
• /
• 1990

튀김유의 품질평가 방법으로서 high performance size exclusion chromatography를 이용하여 이중체, 다중체 등의 중합체를 보다 신속하게 분리할 수 있었다. 단일체 triglyceride는 가열시간이 경과함에 따라 감소하는 경향을 나타났으나, 복합처리제를 처리한 처리구의 경우에는 감소속도가 현저히 늦어지는 경향을 나타냈다. 단일체 triglyceride와 극성지질 함량사이에는 좋은 상관관계를 나타냈으며, 회기직선의 기울기에 있어서는 처리구와 대조구 모두 거의 같은 경향을 나타났다. 이러한 방법에 의하면, 단일체 중성지질 함량이 71% 이하로 저하된 튀김유는 폐기되어야 하며, 이 값은 극성지질 함량 27%에 대응하는 측정값이다.

• Bulletin of the Korean Chemical Society
• /
• 제16권7호
• /
• pp.640-643
• /
• 1995

The internal standard method, which is widely used in chromatographic techniques, is applied to the size exclusion chromatography for the on-line determination of dn/dc value of the sample. This method is found to provide the dn/dc value with suitable accuracy in determining the absolute molecular weight and the molecular weight distribution of the polymers when a light scattering detector is used.

• Macromolecular Research
• /
• 제14권3호
• /
• pp.383-386
• /
• 2006

High temperature size exclusion chromatography (SEC) has been used widely for the characterization of crystalline polymers, for which high temperature operation above the polymer melting temperature is required to dissolve the polymers. However, this high temperature operation has many advantages in SEC separation in addition to merely increasing polymer solubility. At high temperature the eluent viscosity decreases, which in turn decreases the column backpressure and increases the diffusivity of the analytes. Therefore, many reports on the high temperature operation of high performance liquid chromatography (HPLC) have focused on shortening the analysis time and enhancing the resolution. However, the application of high temperature SEC analysis to exploit the merits of high temperature operation is scarce. In this article, therefore, we report on a new apparatus design for high temperature SEC.

• Bulletin of the Korean Chemical Society
• /
• 제32권4호
• /
• pp.1315-1320
• /
• 2011

Whey protein (WP) is a mixture of proteins, and is of high nutritional values. WP has become an important source of functional ingredients in various health-promoting foods. In this study, size-exclusion chromatography (SEC) and asymmetrical flow field-flow fractionation (AsFlFFF) were used for separation and analysis of whey proteins. It was found that a lab-prepared WP from raw milk is mostly of ${\beta}$-lactoglobulin with small amount of higher molecular weight components, while a commercial whey protein isolate (WPI) powder contains relatively larger amount of components other than ${\beta}$-lactoglobulin, including IgG and protein aggregates. Results suggest that AsFlFFF provides higher resolution for the major whey proteins than SEC in their normal operation conditions. AsFlFFF could differentiate the BSA and Albumin, despite a small difference in their molecular weights, and also was able to separate much smaller amount of aggregates from monomers. It is noted that SEC was able to show the presence of low molecular weight components other than the major whey proteins in the WP samples, which AsFlFFF could not show, probably due to the partial loss of those low molecular weight species through the membrane.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제10권2호
• /
• pp.122-127
• /
• 2005

A size exclusion chromatography (SEC) process, in the presence of denaturant in the refolding buffer was developed to refold recombinant human $interferon-\gamma$ ($rhIFN-\gamma$) at a high concentration. The $rhlFN-\gamma$ was overexpressed in E. coli resulting in the formation of inactive inclusion bodies (IBs). The IBs were first solubilized in 8 M urea as the denaturant, and then the refolding process performed by decreasing the urea concentration on the SEC column to suppress protein aggregation. The effects of the urea concentration, protein loading mode and column height during the refolding step were investigated. The combination of the buffer-exchange effect of SEC and a moderate urea concentration in the refolding buffer resulted in an efficient route for producing correctly folded $rhIFN-\gamma$, with protein recovery of $67.1\%$ and specific activity up to $1.2\times10^7\;IU/mg$.