• Title/Summary/Keyword: 이온-크로마토그래피

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Purification and Characterization of $\beta$-Xylosidase B of Bacillus stearothemophilus No.236 Produced by Recombinant Escherichia coli. (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus No.236 $\beta$-Xylosidase B의 정제 및 특성)

  • 장욱진;조쌍구;최용진
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.297-302
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    • 1998
  • $\beta$-Xylosidase B was produced by Escherichia coli HB101/pKMG12 carrying the xylB gene of Bacillus stearothermophilus No.236 on its recombinant plasmid. The $\beta$-xylosidase B produced was purified by ammonium sulfate fractionation, DEAE-Sepharose CL-6B, Sephacryl S-200 and Superdex 200 HR gel filtration. The purified enzyme showed the highest activity at pH 6.5 and 5$0^{\circ}C$. But, the enzyme was observed to be very sensitive to the pH and temperature of the reaction mixture. The enzyme was activated about 35% of its original activity in the presence of 1 mM of $Mn^{2+}$ but it was completely inhibited by $Ag^{+}$, $Cu^{2+}$and $Hg^{2+}$ions. In contrast with the $\beta$-xylosidase A, the B enzyme was found to have $\alpha$-arabinofuranosidase activity though the activity was fairly low compared with the $\alpha$-arabinofuranosidase produced from the arfI gene of the same Bacillus stearothermophilus. Therefore, $\beta$-xylosidase B is considered to be more suitable than $\beta$-xylosidase A at least for the biodegradation of arabinoxylan. The $K_{m}$ and V$_{max}$ values of the $\beta$-xylosidase B for o-nitrophenyl-$\alpha$-D-xylopyranoside were 6.43 mM and 1.45 $\mu$mole/min, respectively. Molecular mass of the enzyme was determind to be about 54 kDa by SDS-PAGE and 160 kDa by Superdex 200HR gel filtration, indicating that the functional $\beta$-xylosidase B was composed of three identical subunits.s.

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An Experimental Study on the Ion Reaction and the Electrochemical Rebar-Corrosion in Aqueous Solution Mixed with Sulfate and Chloride Ion-Reactive Material (황산, 염소이온 반응 소재 혼입 수용액에서의 이온반응성 및 전기화학적 철근 부식에 관한 실험적 연구)

  • Ryu, Hwa-Sung;Shin, Sang-Heon;Kang, Tae-Won;Lim, Chang-Gil;Kim, Hong-Tae
    • Journal of the Korea Institute of Building Construction
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    • v.19 no.1
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    • pp.31-38
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    • 2019
  • In this study, amine derivatives and ion exchange resins were selected to actively control penetration ions ($SO{_4}^{2-}$, $Cl^-$) as the element technology of repair materials for concrete structures in drainage environments. Ions ($SO{_4}^{2-}$, $Cl^-$) adsorption performance and corrosion resistance of calcium hydroxide solution with amine derivative and ion exchange resin were confirmed by ion chromatography and potentiostat analysis. As a result of the experiment, it was confirmed that the amine derivative is excellent in the adsorption of chlorine ion and the ion exchange resin is excellent in the adsorption of sulfate ion. It has been confirmed that corrosion resistance can be increased by proper combination of two materials in the calcium hydroxide solution containing sulfate ion and chloride ion simulating sewage environment.

Evaluation of the Giggenbach Bottle Method with Artificial Fumarolic Gases (인공 분기공 가스를 이용한 Giggenbach bottle 법의 평가)

  • Lee, Sangchul;Kang, Jungchun;Yun, Sung Hyo;Jeong, Hoon Young
    • Journal of the Korean earth science society
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    • v.34 no.7
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    • pp.681-692
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    • 2013
  • We aimed to evaluate the effectiveness of the Giggenbach bottle method and develop the related pretreatment and analytical methods using artificial fumarolic gases. The artificial fumarolic gases were generated by mixing $CO_2$, CO, $H_2S$, $SO_2$, $H_2$, and $CH_4$ gas streams with a $N_2$ stream sparged through an acidic medium containing HCl and HF, with their compositions varied by adjusting the gas flow rates. The resultant fumarolic gases were collected into an evacuated bottle partially filled with a NaOH absorption solution. While non-condensible gases such as CO, $H_2S$, and $CH_4$ accumulated in the headspace of the bottle, acidic components including $CO_2$, $SO_2$, HCl, and HF that were dissolved into the alkaline solution. Like other acidic components, $H_2S$ also dissolved into the solution, but it reacted with dissolved $Cd^{2+}$ to precipitate as CdS when $Cd(CH_3COO)_2$ was added. The non-condensible gases were analyzed on a gas chromatography. Then, CdS precipitates were separated from the alkaline solution by filtration, and they were pretreated with $H_2O_2$ to oxidize CdS-bound sulfide into sulfate. In addition, a portion of the solution was also pretreated with $H_2O_2$ to oxidize sulfite to sulfate. Following the pretreatment, the resultant samples were analyzed for $SO_4^{2-}$, $Cl^-$ and $F^-$ on an ion chromatography. In the meanwhile, dissolved $CO_2$ was analyzed on a total organic carbon-inorganic carbon analyzer without such pretreatment. According to our experimental results, the measured concentrations of the fumarolic gases were shown to be proportional to the gas flow rates, indicating that the Giggenbach bottle method is adequate for monitoring volcanic gas. The pretreatment and analytical methods employed in this study may also enhance the accuracy and reproducibility of the Giggenbach bottle method.

A Study on the Analysis of Methylprednisolone Acetate and its Metabolites in Rat Urine by LC/MS (LC/MS를 이용한 뇨중에서의 Methylprednisolone Acetate 및 그 대사물질 분석에 관한 연구)

  • Park, Song-Ja;Pyo, Hee Soo;Kim, Yun Je;Park, Seong Soo;Park, Jongsei
    • Analytical Science and Technology
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    • v.8 no.2
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    • pp.139-159
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    • 1995
  • Positive ion mass spectra of some corticosteroids were obtained by using liquid chromatography-mass spectrometry(LC-MS). The base peak of each compound showed the protonated molecular ion [$MH^+$], ammonium adduct ion [${MNH_4}^+$] or [$MH^+-60$] ion according to its chemical structure and other characteristic mass ions were [$MH^+-18$], [${MNH_4}^+-18$] and so on. Several rat urinary metabolites of methylprednisolone acetate after the oral administration were detected by the thermospray LC-MS. The identified major metabolites were 20-hydroxymethylprednisolone(20-HMP), methylprednisolone(MP) and methylprednisone(11-KMP), which were supposed to be formed by deacetylation at the position of C-21, reduction at C-20, oxidation at C-11, or due to the bond cleavage between C-17 and C-20.

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Structural determination of triterpenic acids in Prunellae Spica by fast atom bombardment tandem mass spectrometry (하고초의 생리활성 성분 Triterpenic Acids의 FAB-MS를 이용한 구조 규명)

  • Ahn, Young Min;Lee, Kang Ro;Hong, Jongki
    • Analytical Science and Technology
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    • v.21 no.4
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    • pp.245-258
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    • 2008
  • Five triterpenic acids as marker compounds were extracted and isolated from Prunellae Spica by column chromatography and reversed-phase high-performance liquid chromatography (HPLC), and their purity was determinated by HPLC (purity ${\geq}90%$). Molecular weight and elemental compositions of the five marker compounds were determined by fast atom bombardment high-resolution mass spectrometry (FAB-HRMS). The structural determination of the five marker compounds was carried out fast atom bombardment collision-induced dissociation tandem mass spectrometry (FAB-CID-MS/MS). The collision-induced dissociation (CID) of protonated molecules $[M+H]^+$ and deprotonated molecules $[M-H]^-$ produced diverse product ions due mainly to retro Diels-Alder reaction (RDA), dehydration and decarboxylation. Moreover, the CID-MS/MS spectra of the $[M-H]^-$ ions were observed charge-remote fragmentation (CRF) patterns. On the basis of interpretation of CID-MS/MS spectra, structural elucidation of triterpenic acids isolated from Prunellae Spica was clearly performed.

A study on the derivatization technique for tamoxifen metabolites in human urine by gas chromatography/mass spectrometry (기체크로마토그래피/질량분석기를 이용한 인체 내 뇨시료에서의 Tamoxifen 대사체 검출을 위한 유도체화 연구)

  • Kim, Yunje;Lee, Yoonjung
    • Analytical Science and Technology
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    • v.17 no.4
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    • pp.322-336
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    • 2004
  • The improved derivatization technique of tamoxifen metabolite in human urine is described for the acylation method that they are substituted by derivatization reagent like acyl anhydride for use of gas chromatography/mass spectrometry. The hydroxyl group of tamoxifen metabolite was derivatized by trifluoroacetic anhydride (TFAA), pentafluoroacetic anhydride (PFPA) and heptaflorobutylic anhydride (HFBA). It was investigated to the gas chromatography/mass spectrometry (GC/MS) technique use negative ion chemical ionization (NCI), positive ion chemical ionization (PCI) and electron impact (EI). In acylation of the metabolites of tamoxifen, the effective reaction temperature and time were shown to be at $50^{\circ}C$ for 30 min. The 4-hydroxytamoxifen, which is known to major metabolite of tamoxifen, was not detected in human urine, whileas the hydroxymethoxytamoxifen was detected. We thought that this result was from the single dose of tamoxifen.

Improved Procedure for Purification of Clostridium botulinum type B Toxin (Clostridium botulinum Type B 독소의 정제방법에 관한 연구)

  • 박문국;양규환
    • Korean Journal of Microbiology
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    • v.20 no.4
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    • pp.183-188
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    • 1982
  • The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163, 000. Subunits of 106, 000 and 56, 000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

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세포능식촉진인자(MPF)의 특이적 억제제 생산 토양균주의 검색 및 억제제의 분리와 특성 규명에 관한 연구

  • 박희동;박상곤;이승기
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.258-258
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    • 1994
  • 본 연구에서는 토양균이 생성하는 2차 대사산물로부터 세포증식 촉진인자 (Maturation Promoting Factor. MPF)인 cdc2/cdk2-cyclin의 복합체를 특이적으로 억제하는 물질을 분리하고 그 물질의 생리활성을 조사하였다. 토양으로부터 순수분리된 300여개의 토양균에서 생성된 배양액을 취하여 MPF의 특이적인 기질인 합성 peptide (CSH103 :HATPPKKKRK)를 사용하여 인산화 활성을 측정하였다. 그중 MPF 활성 억제능이 90% 이상인 19개의 균주를 1차적으로 선정한 후 각각에 대하여 열/pH에 대한 안정성, 각종 용매에 대한 추출성 등 이화학적 성질을 규명하였으며 이중 균주 LPL 931로부터 MPF 활성 억제제를 분리하고자 하였다. 예비실험의 결과로부터 토양균 LPL 931을 대량배양하여 열처리하고, 비이온성 수지인Amberlite XAD-2에 결합시키고 70% acetone으로 용출시켰다. 이 추출물로부터 ethylacetate와 n-butanol을 사용하여 MPF 억제 활성물질을 추출하였다. 이 추출액을 실리카겔 관 크로마토그래피, 분취 TLC, 분취 HPL를 하여 MPF 활성 억제 분획을 분리하였다.

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Quantitative Analysis of Nitrate and Nitrite in Rain, Milk and Infant Formula using Ion Chromatography (이온 크로마토그래피를 이용한 빗물과 분유, 우유중의 nitrite와 nitrate의 정량분석)

  • Kang, Mi-Jeong;Lee, Sueg-Geun
    • Analytical Science and Technology
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    • v.12 no.6
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    • pp.583-586
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    • 1999
  • The quantitative analysis of nitrate and nitrite in rain, milk and infant formula was done by Ion Chromatography. The nitrite was not detected(<0.1 mg/L) in all the samples. However, the nitrate was detected in the range of 0.1~4.9 mg/L in rain, 9.8~19.8 mg/L in milk, and 80~300 mg/L in infant formula, respectively. Some content of nitrate is close to the maximum contaminant level(MCL) which is 10 mg/L as $NO_3-N$, 44.3 mg/L as $NO_3$.

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Determination of Adsorption Isotherms and Separation of L-arabinose and D-ribose in Cation Exchange Chromatography and HPLC (양이온 교환 크로마토그래피와 HPLC에서의 L-arabinose와 D-ribose의 분리 및 등온 흡착곡선 결정)

  • Jeon, Young-Ju;Kim, In-Ho
    • 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.