• 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.

• Analytical Science and Technology
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• v.25 no.6
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• pp.483-491
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• 2012

This study is to investigate the issues on how to secure stability during the purification process for the production of recombinant protein A. The final recombinant protein A is produced by passing through the cation exchange column (SP) and the anion-exchange column (Q) during the production process, for which the samples produced by the step-by-step processes can be exposed to trouble in securing stable storage in case the next process cannot be taken within the proper time period. Accordingly, this study aims to evaluate the proper storage conditions and length of time when storing samples produced in the production process. That is, in this study, how to store fair samples, how long the storage period should be set up, and how to evaluate the security of its quality depending on time are dealt with. The items to be experimented with were enodotoxin, SDS-PAGE, HPLC purity and concentration. Experimental results showed that after passing the cation exchange column, when stored at $4^{\circ}C$ or room temperature, SDS-PAGE showed a major band, endotoxin is 5.0 Eu/mg or less, and concentration is on average of 8.21 to 8.24 mg/mL and RSD% 0.10~0.62%. In addition, HLPC purity showed somewhat stable results; at the HPLC purity 214 nm, the average is 99.24% to 99.37% and RSD% is 0.22~0.29%, while the average is 89.72% to 89.80% and RSD% 0.62~1.26% at 280 nm. On the contrary, after passing the anion exchange column, when stored at $4^{\circ}C$ or room temperature, SDS-PAGE revealed the major band, endotoxin is 0.5 Eu/mg or less, and concentration is on average of 5.59 mg/mL and RSD% 0.03~0.10%. when it comes to HLPC purity, the result showed that at the HPLC purity 214 nm, the average is 99.74% and RSD% is 0.10~0.11%, while the average is 96.16% to 96.85% and RSD% 0.72~1.13%. In conclusion, the stability of fair samples of recombinant protein A during the manufacturing process could be obtained without substance decomposition for 7~8 days at $4^{\circ}C$ or 20~21 days at room temperature.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1028-1032
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• 2003

To assess accurate methods for measuring sulfur dioxide residue in pickles, the acid distillation HPLC-UV method and Monnier Willams modified method were examined. By the acid distillation HPLC-UV method, sulfites released from pickles by acid distillation were absorbed in 1% triethanolamine solution and detected as sulfite ion by HPLC with UV monitoring at 240nm. An anion exchange column was employed with 1.8mM $Ma_2CO_3-1.7mM\;NaHCO_3$ solution as a mobile phase, $84.0{\sim}91.7%$ of sulfite added to pickled radish were recovered. Total sulfite levels from 48 kinds of pickles analyzed by acid distillation HPLC-UV was compared with those analyzed by the Monnier Williams modified method. The Monnier Williams modified method showed higher levels of sulfur dioxide than the acid distillation HPLC-UV method due to the presence of volatile acids in pickles. The concentration of sulfur dioxide was in the range of $N.D{\sim}173.05ppm$ in pickled radish and over 30ppm of sulfur dioxide from 3 samples by the acid distillation-HPLC-UV method.

• Journal of the Korean Chemical Society
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• v.17 no.6
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• pp.428-433
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• 1973

The quantitative separations of a mixture containing equal amount of each anion such as Si(IV), As(V), P(V), S(VI), W(VI) and Cr(VI) are carried out by the elution through 20${\times}3.14cm^2$ column of anion exchange resin, Dowex 1${\times}$8. The eluents are a mixture of 0.07 M hydrochloric acid and 0.03 M sodium chloride (pH = 1.30) for Si(IV), As(V) and P(V) species, a mixture of 0.6 M sodium chloride and 0.3 M sodium hydroxide for S(VI), W(VI) and Cr(VI) species, and 0.1 N sodium sulfite (pH = 3.48) for P(V) and As(V) species. The subsidiary anions in a standard mixture such as Si(IV), As(V), S(VI), P(V) and W(VI) are separated together from large amount of Fe(III) by the elution through 30cm${\times}3.14cm^2$ column of the resin, Dowex${\times}$50w${\times}$12, using a mixture of 0.1 M sodium nitrate and 2 percent dimethylsulfoxide aqueous solution as an eluent. Si(IV), As(V), S(VI), P(V) and W(VI) eluted together are separated quantitatively under the same conditions as in the separations of the anion mixture. By the conditions obtained in the separations of the standard mixture, Fe(III) and all of the subsidiary anions in steel are quantitatively separated.

• Korean Journal of Agricultural and Forest Meteorology
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• v.2 no.4
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• pp.190-197
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• 2000

Anions such as C $l^{[-10]}$ , N $O_3$$^{[-10]}$ , S $O_4$$^{2-}$, P $O_4$$^{3-}$, and organic anions, that do not become a part of the clay mineral crystal lattice, are of considerable interest in soils which are a potential sink caused by acid rain. In this paper, elution of native sulfate and breakthrough curves (BTC) were obtained from miscible displacement of non-specifically or specifically adsorbed anions through non-saturated or saturated Bt soil of Chungwon series. The shape and position of the BTC's could be affected by adsoprtion and ion exchange onto the soil particle surfaces. Measured BTC's for oxalic acid under unsaturated and saturated conditions showed that less pore volumes were required to displace the native S $O_4$$^{2-}$S from the soil column, and that maximum detection limit of oxalic acid reached earlier than under unsaturated. The retarded BTC's to the righthand side could be attributed by different adsorption behavior of each anion, although BTC's may be influenced by the smaller order of velocity change. The alternate breakthrough and elution curves show the rapid approach to the maximum detection limit of C/Co = 1, compared to progressive tailing of elution curve to reach to C/Co = 0. The probable explanation for asymmetric elution patterns for both anion is that the anion was selectively adsorbed on the positively charged soil surface from the solution passing in the soil column. On the other hand, the variations of pH in effluent showed that pH was increased to 7 in the first 6 pore volume and then gradually decreased to pH 4.

• Analytical Science and Technology
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• v.10 no.1
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• pp.53-59
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• 1997

Chloride form anion exchange resin was used to separate one of the elements from the rare earth mixture using respectable Ln-EDTA eluent. $Sm^{3+}$, $La^{3+}$ or $Ce^{4+}$ complexed with EDTA was passed through the resin column and eluted with a Sm-EDTA solution as an eluent. Here all the rare earth element ions except $Sm^{3+}$ were passed. Adsorbed $Sm^{3+}$ in resin was eluted with 1.0 M HCl solution. If La-EDTA solution as an eluent was used to separate lanthanum ions, lanthanum ions were eluted together with other rare earth elements. When Ce-EDTA solution was also used for separation of $Ce^{4+}$, it was eluted in the region of other rare earth elements. In the case of Sm-EDTA elution, the elution mechanism was as follows : Absorption : $RCl+Ln-Y^-{\leftrightarrows}RLnY+Cl^-$, Sm-EDTA elution : $RLnY+Sm-Y^-{\leftrightarrows}RSmY+Ln-Y^-$, HCl elution : $RSmY+HCl{\leftrightarrows}RCl+Sm-Y^-$.

• Korean Journal of Pharmacognosy
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• v.32 no.1 s.124
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• pp.31-38
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• 2001

A lectin was purified from Allomyrina dichotoma (ADL) by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. Several biochemical properties of ADL were characterized as follows: ADL from gel filtration column chromatography showed single band on SDS-PAGE. ADL agglutinated the erythrocytes of rabbit and human A, B, O, AB. Agglutinability was relatively stable at basic pH, and was stable at temperature below $40^{\circ}C$. Agglutinability was not affected by metal ions and EDTA. This lectin was proved to be a glycoprotein which contains 0.47% of sugars. The molecular weight of ADL was estimated to be 97,000 dalton by SDS-PAGE. By amino acid analysis, ADL exhibited high amounts of aspartic acid. The lectin's immunomodulating effect was measured as cytokine production. The productions of 5 cytokines $(IL-1{\alpha},\;IL-2,\;IL-6,\;IFN{\gamma}\;and\;TNF{\alpha})$ from peripheral blood mononuclear cells were measured by ELISA. The lectin induced the highest secretion of IL-2 at 8 hr, $TNF{\alpha}$ at 4 hr, and $IFN{\gamma}$ at 24hr, respectively. These results suggest that ADL can elicit the production of detectable cytokines from PBMC.

• Bulletin of the Korean Chemical Society
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• v.34 no.12
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• pp.3817-3824
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• 2013

Various separation modes in HPLC, such as anion exchange (AE), reversed-phase (RP), and affinity (AF) chromatography were examined for the separation of selenium species in human blood serum and urine. While RP- and AE-HPLC were mainly used for the separation of small molecular selenium species, double column AF-HPLC achieved the separation of selenoproteins in blood serum efficiently. Further, the effluent of AF-HPLC was enzymatically hydrolyzed and then analyzed with RP HPLC for selenoamino acid study. The versatility of the hybrid technique makes the in-depth study of selenium species possible. For quantification, post column isotope dilution (ID) with $^{78}Se$ spike was performed. ORC ICP/MS (octapole reaction cell inductively coupled plasma/mass spectrometry) was used with 4 mL $min^{-1}$ Hydrogen as reaction gas. In urine sample, inorganic selenium and SeCys were identified. In blood serum, selenoproteins GPx, SelP and SeAlb were detected and quantified. The concentration for GPx, SelP and SeAlb was $22.8{\pm}3.4\;ng\;g^{-1}$, $45.2{\pm}1.7\;ng\;g^{-1}$, and $16.1{\pm}2.2\;ng\;g^{-1}$, respectively when $^{80}Se/^{78}Se$ was used. The sum of these selenoproteins ($84.1{\pm}4.4\;ng\;g^{-1}$) agrees well with the total selenium concentration measured with the ID method of $87.0{\pm}3.0\;ng\;g^{-1}$. Enzymatic hydrolysis of each selenium proteins revealed that SeCys is the major amino acid for all three proteins and SeMet is contained in SeAlb only.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.5
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• pp.1321-1326
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• 2006

Total ginsenosides were purified and their antioxidant, antibacterial and anticancer activities were measured. The crude extracts of ginseng, which were extracted with 75% ethanol by ultrasonification method, were firstly purified on AB-8 macroporous adsorption column to remove water soluble impurities, and decolored on Amberlite IRA 900 Cl anion-exchange column. Then, they were purified on Amberlite XAD16 adsorption column to delete the non-polar impurities. Total ginsenosides contents of the purified extracts were 79.4%, 71.7% and 72.5% in cultured wild ginseng, red ginseng and white ginseng, which were significantly increased than those of crude extracts. All of the three extracts showed concentration-dependant scavenging activities against DPPH radicals, among which white ginseng showed the most powerful activity. Cultured wild ginseng roots showed strongest effect against both B. subtilis PM 125(Gram-positive) and E. coli D31 (Gram-negative) bacteria, while red ginseng and white ginseng only showed the activity against B. subtilis. According to the result of the MTT assay, ail of the three extracts inhibited the growth of U-937 human hohistiocytic lympma cell, which were significantly different (p < 0.05) when compared to the control.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.136-140
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• 2000

A sensitive and quantiative method for the structural analysis of oligosaccharide was established for the glycoform analysis of glconproproteins. Inthis study, n-linked oligosaccharides of human IgG and bovine transferin were analyzed for the evaluation of the methydrate moiety ofthe method. Chrbohydrate moiety of glycoprotein was relased by hydrazinolysis and purified by paper chromatography. The oligosaccharides were labeled with a fluorescent bye, 2-aminobenzamide, for the enhancement of detection sensitivity. sialylated (acidic) oligosaccharides were separated from neutral oligosaccharide by employing a strong anion-exchange column(MonoQ) followed by the treatment with sialidase. Enzymatically desiayated fractions and neutral fractions of oligosaccharides were applied to normal-phase HPLC to resolve the peaks according to glucose unit (GU). The structure of separated molecules was further determined by sequential digestion with exoglycosidases. As a result, disialylated biantennary complextype oligosaccharide was found to be a major sugar chain in bovine transferrin (63%). In human IgG, core fucosylated asialobiantennary complex oligosaccharides were dominant. These results coincided well with reported results.