• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.4
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• pp.562-566
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• 2003

A $\beta$-mannanase of Bacillus sp. was purified by DEAE Sephacel ion exchange column chromatography. The specific activity of the purified enzyme was 17.41 units/mg protein, representing an 84.74-folds purification of the original crude extract. For the separation of two types of hydrolysates by the action of purified $\beta$-mannanase, carbon column chromatography, sephadex G-25 column chromatography and thin layer chromatography were accomplished. Main hydrolysates were D.P value 5 and 7 containing of low D.P values. By the method of FACE (Fluorophore Assisted Carbohydrate Electrophoresis), two types of hydrolysates were identified to homo type.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.4
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• pp.285-291
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• 2004

A simulated moving bed (SMB) chromatography system is a powerful tool for preparative scale separation, which can be applied to the separation of chiral compound. We have de-signed our own lab-scale SMB chromatography using 5 HPLC pumps, 6 stainless steel columns and 4 multi-position valves, to separate a racemic mixture of ketoprofen in to its enantiomers. Our design has the characteristics of the low cost for assembly for the SMB chromatography and easy repair of the unit, which differs from the designs suggested by other investigators. It is possible for the flow path through each column to be independently changed by computer control, using 4 multi-position rotary valves and 5 HPLC solvent delivery pumps. In order to prove the operability of our SMB system, attempts were made to separate the (S)-ketoprofen enantiomer from a ketoprofen racemic mixture. The operating parameters of the SMB chromatography were calculated for ketoprofen separation from a batch chromatography experiment as well as by the triangle theory. With a feed concentration of 1 mg/mL, (S)-ketoprofen was obtained with a purity of 96% under the calculated operating conditions.

• Korean Journal of Food Science and Technology
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• v.31 no.5
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• pp.1386-1391
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• 1999

The methanol extract of Gardenia jasminoides Ellis showed antimicrobial activity against bacteria and yeasts. The extract was successively purified with solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, octadecylsilane column chromatography. The purified active substance was isolated by high performance liquid chromatography. The isolated compound was 3,4-dihydroxybenzoic acid which was determined by mass spectrometer, gas chromatograph-mass spectrometer, $^{1}H-nuclear$ magnetic resonance, $^{13}C-nuclear$ magnetic resonance and two-dimensional nuclear magnetic resonance. The content of 3,4-dihydroxybenzoic acid was $32.7\;{\mu}g/g$ in dried fruits of Gardenia jasminoides Ellis.

• Korean Journal of Veterinary Research
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• v.13 no.1
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• pp.23-30
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• 1973

Thin layer, paper and column chromatography were compared for the separation, detection and quantitation of three kinds of estrogen in urine of dairy cows. While thin layer chromatography utilizing silica gel was better for the detection of estrogens, column chromatography using celite 545 was preferable. Spectrophotometry was compared with fluorometry for determination of estrone, estradiol-17 ${\beta}$ and estriol eluted by paper chromatography and column chromatography. Optical density of three standard estrogens showed almost same curve at maximum absorption wave length of 230 and $282m{\mu}$. However, the former showed a higher peak. In fluorometry, the fluorescence intensity of estrone and estradiol-17 ${\beta}$ were rather strong, when the estrogens were dissolved in sulfuric acid, and showed higher sensitivity than that of the spectrophotometry. However, in the case of estriol was exceptional.

• FOCUS: LIFE SCIENCE
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• no.1
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• pp.6.1-6.9
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• 2024

The document is a white paper on Hydrophilic Interaction Liquid Chromatography (HILIC) analysis method development. HILIC is a type of chromatography that uses an organic/aqueous mobile phase and a polar stationary phase. In HILIC, water is a strong solvent, and unlike in Reversed Phase Liquid Chromatography (RPLC), increasing the proportion of water in the mobile phase reduces the retention time of the analyte. The paper discusses when to consider HILIC analysis methods, the advantages of HILIC, and the challenges often encountered due to the lack of understanding of HILIC mechanisms compared to RPLC. It also provides a systematic flowchart for intelligent solutions for HILIC analysis method development, which includes a three-step approach for chromatography analysis method development. The first step involves gathering as much information as possible about the analyte (e.g., pKa, log P, log D). The second step involves analyzing the sample under different pH conditions using three HILIC columns in either isocratic or gradient mode to identify the suitable column/pH combination for the analyte. The third step involves optimizing the separation by investigating other parameters such as temperature and ionic strength, and assessing the robustness of the method. The paper emphasizes that the selection of the appropriate stationary/mobile phase combination, based on the differences between the HILIC stationary phases and the mobile phase pH, can provide high selectivity in the analysis. This step-by-step approach can help users develop an efficient analysis method.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.1025-1032
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• 2005

This study was carried out to separate ovotransferrin in chicken egg white by gel chromatography and heparin affinity chromatography. In gel filtration which was performed with 50mM Phosphate buffer (pH 7.2, 0.15M salt) at a flow rate of 2.0 ml/min, ovotransferrin and ovalbumin were eluted together in fraction number 11-16. In order to separate pure ovotransferrin, fraction No. 12-14 of them which have high concentration of ovotransferrin were concentrated and rechromatographed. However, the ovotransferrin did not separated clearly. In heparin affinity chromatography, the separation was performed with 50mM ethylaminetetraacetic acid (EDTA, pH7.2) and 50mM Phosphate buffer (pH 7.2, 0.15M salt contained) on ferrous and ferric ion saturated column at as same flow rate as gel filtration system's. Ovotransferrin and albumin were eluted together at 10-15min (fraction No.3) and 15-20min (fraction No.4), respectively. However, purified ovotransferrin was eluted at 156-165min and 165-175min (tube No.32-33) with 50 mM phosphate buffer (pH 7.2, 0.15M salt free), respectively. Heparin affinity chromatography with ferric ion saturated column was resulted in the best separation of ovotransferrin rather than separation by gel chromatography and ferrous ion saturated heparin affinity chromatography.

• Journal of Ginseng Research
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• v.13 no.2
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• pp.254-259
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• 1989

A radioprotective ginseng protein fraction was obtained from Korean white ginseng powder by the following isolation and purification procedures: Tris-HCI buffer extraction, 70% ammonium sulfate fractionation, CM-rellulosr column chromatography, heat inactivation and Sephadex G-75 column chromatography. This fraction was further purified by Sepharose 4B and Sephadex G-150 column chromatographies. Three fractions obtained were subjected to Native-PAGE and SDS-PAGE using gradient gels and the silver staining method. Molecular weights of the native proteins and their subunits were estimated.

• Korean Journal of Microbiology
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• v.29 no.2
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• pp.92-96
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• 1991

The soluble cytochrome c-551 of photosynthetic bacterium, Rhodopseudomonas gelatinosa ATCC 17013 was purified through a sequene of four step chromatography including CM-cellulose ion-exchange chromatography, DEAE-Sephacel chromatography, Sephacryl s-200 gel permeation chromatography, and HPLC (SP-5PW). The molecular weight of the purified cytochrome c-551 was 14, 600 Da, and this protein shows the absorption peak at 551 nm, 522 nm, and 417 nm as the reduced form, and at 412 nm as the oxidized form. The cytochrome c-551 seems to be a substrate for the terminal oxidase in the electron transport chain.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.390-393
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• 1990

A cellobiohydrolase was purified from the culture broth of Trichoderma uiride by using immunoaffinity chromatography. A single protein band in polyacylamide gel electrophoresis and isoelectrofocusing after immunoaffinity purification corresponded to cellobiohydrolase activity. A immunoaffinity purified cellobiohydrolase is more effective in the hydrolysis of highly crystalline cellulose than amorphous cellulose.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.147-150
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• 2000

Bioprocess chromatography is probably the most widely used unit operation in biopharmaceutics manufacturing. To obtain a good quality product reproducibly, the chromatography process validation should be performed carefully. We developed a miniaturized automatic chromatography system, MiniValChrom, for column performance validation.