• Journal of Food Hygiene and Safety
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• v.32 no.5
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• pp.404-410
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• 2017

An analytical method for determination of propylene glycol alginate (PGA) in food products was developed by HPLC-size-exclusion chromatography. The GF-7M HQ column and LT-ELSD detector were determined by considering the instrumental analysis conditions for PGA analysis. The pretreatment method for the analysis of PGA was suitable for 3 hr extraction at $20^{\circ}C$ and 150 rpm according to the extraction temperature. Linearity ($R^2$) for the analysis of PGA was 0.9873 at calibration curve range of 300, 500, 700, 1,000, and 1,500 mg/kg (5 points). The limit of detection and limit of quantification of PGA on HPLC system was 171.43 and 519.50 mg/kg, respectively. The accuracy and coefficient of variation obtained by size-exclusion chromatography were 86.1~110.4% and 4.1~13.5%, respectively. By applying the HPLC-size-exclusion chromatography system, it was possible to analyze the contents of PGA in 134 different types of food products.

• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.416-423
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• 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.

• Analytical Science and Technology
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• v.8 no.4
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• pp.405-410
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• 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.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.09a
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• pp.9-17
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• 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
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• v.1 no.2
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• pp.87-91
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• 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.

• Korean Journal of Food Science and Technology
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• v.22 no.1
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• pp.33-37
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• 1990

A simple and rapid method, based on separation of a flying oil into monomeric, dimeric and oligomeric triglyceride by means of high performance size exclusion chromatography on Ultrastyragel column (500 A), is proposed for evaluation of the quality of used frying oil. The relative area of the monomeric triglyceride was decreased with frying time increase, however, the decrease rate was significantly reduced by treatment of a composited powder (porous rhyorite/citric acid 40/56/4). This measurement showed good linear relationship with change in polar component measurement. There were no significant differences between the slopes of regression lines in both treated and non-treated frying oil system for relationship between monomeric triglyceride and polar component. By this method, it was found that a frying oil habe to be discarded if the content monomeric triglyceride decreased to 71%, which was corresponed to 27% polar component.

• Bulletin of the Korean Chemical Society
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• v.16 no.7
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• pp.640-643
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• 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
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• v.14 no.3
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• pp.383-386
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• 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
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• v.32 no.4
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• pp.1315-1320
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• 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
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• v.10 no.2
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• pp.122-127
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• 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 $1.2\times10^7\;IU/mg$.