• Title/Summary/Keyword: column chromatography technique

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High-performance liquid chromatographic assay and oral pharmacokinetics of new anti-HIV uracil derivatives, KR-V analogues, in rats

  • Lee, Youngmi;Heeok Shim;Seoungryong Yu;Hochul Shin;Moonkoo Chung;Jungkoo Roh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.123-123
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    • 1998
  • A number of uracil derivatives have been developed as anti-AIDS drugs having a mechanism of inhibiting cellular reverse transcriptase. A simple and rapid assay technique for recently synthesized KR-V analogues was developed using a high-performance liquid chromatography, and oral pharmacokinetics was examined for assessing their oral bioavailabilites. Plasma samples were analyzed by reversed-phase HPLC using an ODS column with an ultraviolet detection system. All the analogues were eluted within 12 min and the LOQ was 15-30 ng/$m\ell$. The extraction recoveries were higher than 85%, except KR-V1039, 1068 and 1720 having ester group. This chromatic method was well applied to the kinetic studies for KR-V analogues. Among 16 analogues tested in the present work, the 6 compounds including KR-V1123, 1122, 1784, 1783, 1736 and 1700 were found to be bioavailable for oral administration to rats.

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Purification and Enzymatic Properties of Cyclodextrin Glucanotransferase from Bacillus macerans Cultivated in Wheat-bran Medium (밀기울배지를 이용한 Bacillus macerans의 Cyclodextrin Glucanotransferase 생산과 효소특성)

  • 선우양일;안태진
    • KSBB Journal
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    • v.9 no.5
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    • pp.499-505
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    • 1994
  • Bacillus macerans cyclodextrin glucanotransferase(EC 2.4.1.19: 1, 4-${\alpha}$-D(1, 4-${\alpha}$-glucano)-transferase, CGTase) was purified by the technique of starch adsorption and DEAE-cellulose column chromatography. The molecular weight of the enzyme was 67,000, consisting of a subunit. The enzyme converted starch into ${\alpha}$-, ${\beta}$-, and ${\gamma}$-CD in the relative amounts of 1:1.68:0.32, respectively. In the early reaction period, maltohexose was formed mainly by the coupling reaction of ${\alpha}$-CD with D-glucose and then other oligosaccharides. Maltotetrose was formed mainly from ${\alpha}$-CD in the initial stage of hydrolysis of the enzyme and then small amount of other oligosaccharides. Maltotriose was a good substrate for the enzyme and maltosyl or D-glucopyranosyl group can be transfered from this sugar. In this work, D-glutosyl transfer was premiered.

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Identification of IY81149 and Its Metabolites in the Rat Plasma Using the On-Line HPLC/ESI Mass Spectrometry

  • Myung, Seung-Woon;Min, Hye-Ki;Jin, Chang-Bae;Kim, Myung-Soo;Lee, Seung-Mok;Chung, Gi-Ju;Park, Seong-Jun;Kim, Dong-Yeon;Cho, Hyun-Woo
    • Archives of Pharmacal Research
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    • v.22 no.2
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    • pp.189-193
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    • 1999
  • Reversed-phase high-performance liquid chromatography/mass spectrometry (HPLC/MS) with an eletcrospray ionization (ESI) interface was applied to the identification of metabolites of IY81149 in the rat plasma. Fragments obtained using collision-induced dissociation (CID) in both positive and negative modes were utilized to elucidate the structure of metabolites. The eluent from the conventional HPLC column was split and directly introduced into an ESI-mass spectrometer for the identification of the structures. the CID technique allowed the sensitive identification of sulfonyl-IY81149 and hydroxy-IY81149 from the rat plasma.

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Immunological Characteristics of the Vitellogenin Induced by $Estradiol-17\beta$ in Male Spotted Flounder, Verasper variegatus (범가자미, Verasper variegatus 수컷에서 $estradiol-17\beta$에 의해 유도된 vitellogenin의 면역학적 특성)

  • KIM Yoon;KIM Woo Jin;BAEK Hea Ja;KIM Kyung Kil;BANG In Chul;HAN Chang Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.480-487
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    • 1997
  • Vitellogenin (Vg) was purified from plasma of $estradiol-17\beta(E_2)-treated$ spotted flounder, verasper variegatus, by preripitation with cold distilled water, followed by fractionation using a Sepharose CL-6B column chromatography. $E_2-induced$ protein was identified as Vg by SDS-PAGE and western blot analysis. The molecular weight of the purified Vg was estimated 175 kD as determined by SDS-PAGE. In order to measure the Vg level, monoclonal antibodies against Vg were produced by hybridoma technique. Purified Vg was immunized into Balb/c mice and then the spleen cells from mice were fused with NS-1 myeloma cells. The hybridoma cells were screened by enzyme-linked immunosorbent assay (WLISA) and subcloned by limiting dilution. The hybridoma clones which secreted antibodies highly reactive to the purified Vg were designated as 4D6. Its specificity was demonstrated by western blot from plasma of untreated, $E_2-treated$ male fish, and purified Vg.

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The Study for Identification of waterborne Spilled Oil by Fast Gas Chromatography (Fast GC를 이용한 해상유출유 감식ㆍ분석 기법 연구)

  • Chung J. W.;Lee W.S.;Yoon J. Y.;Kim H. G.
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.7 no.3
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    • pp.122-130
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    • 2004
  • Crude oil is complex mixture of thousands of different organic compound formed from a variety of organic materials that are chemically converted under differing geological conditions over long periods of time. Also oil composition varies according to crude source, refining, processing, handling and storage. The oil fingerprint method is application of specific knowledge of petrochemicals and use of sophisticated analytical equipment and techniques to identify the source(s) of oil pollution. KNMPA currently utilizes three primary analytical techniques: Gas Chromatography (GC), Fluorescence Spectroscopy(FL) and Infrared Spectroscopy(IR). Of all these techniques, GC technique are most widely used. Gas Chromatography is used as a primary analytical method because high reliableness, high separating efficiency and repeatability, but it is timeconsumable. The study results of identification of waterborne spilled oil by Fast Gas Chromatograph method showed that analytical time is cut down to 30minutes in comparison with packed column method and chromatograms represent high resolution and high repeatability.

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Analysis of selenoaminoacids and selenoproteins in blood serum of sows fed by selenium fortified feed (셀레늄강화 사료를 먹인 모돈 혈청에서의 셀레노아미노산 및 셀레노단백질 분석)

  • Park, Myungsoon;Lee, Sung Hoon;Pak, Yong-Nam
    • Analytical Science and Technology
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    • v.28 no.3
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    • pp.196-203
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    • 2015
  • Selenium species (inorganic selenium, selenoaminoacids, and selenoproteins) were analyzed using anion exchange and affinity chromatography, which were connected to ICP/MS for the blood serum of sows fed by seleniumfortified feed. The Anion Exchange PRP X-100 column was used for the analysis of inorganic selenium (Se4+ and Se6+) and selenoaminoacids. The HEP column was used to separate SelP from GPx+SeAlb in selenoproteins. A quantitative analysis was performed using the post-column isotope dilution technique. The lactating sows were divided into three groups and fed by selenium fortified feed (organic 0.3 mg/kg, 0.6 mg/kg and inorganic 0.6 mg/kg) for four weeks. The test groups showed increases in selenoaminoacids compared with the control group, except the inorganic feed group. There was no significant difference between the organic feed groups. All test groups showed increases in selenoproteins. In particular, SelP showed a large increase that was 1.5 times higher than the other proteins.

Determination of 8-iso-PGF as Oxidative Stress Marker in Human Urine by High Performance Liquid Chromatography with Tandem Mass Spectrometry (LC/MS/MS를 이용한 산화성 스트레스 지표로써 소변 중 8-iso-PGF 분석)

  • Kho, Young-Lim;Lee, Eun-Hee;Chae, Hong-Jae;Choi, Kyung-Ho;Paik, Do-Myung
    • Journal of Environmental Health Sciences
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    • v.36 no.1
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    • pp.44-51
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    • 2010
  • This study aimed to develop analytical method for 8-isoprostanes as biomarkers for oxidative stress with LC/MS/MS technique and to apply the method for human urine samples. Analyzed compounds for urinary oxidative stress markers were 7 stereo-isomers of prostaglandins and the internal standard (iso-$PGF_{2{\alpha}}-d_4$) was used to adjust the recovery rate. The method for determining urinary iso-$PGF_{2{\alpha}}$ consisted of solid phase extraction and LC/MS/MS detection. Separation of isomers of prostaglandins completed by porous graphitic carbon column and buffer solution. Detection limits for urinary markers of oxidative stress, iso-$PGF_{2{\alpha}}$ with LC/MS/MS were 0.01 ng/ml by S/N ratio 3 and 0.028 ng/ml by calculated as to FDA method. The recovery (92.8~101.9%) and precision (8.8~20.7%) of analysis were feasible for detecting iso-$PGF_{2{\alpha}}$ in real human urine samples. We detected 4 isomers of prostaglandins in human urine samples. Mean (standard deviation) of urinary iso-$PGF_{2{\alpha}}$ concentration were 0.231 (0.117) ng/mg creatinine for smoking group and 0.154 (0.082) ng/mg creatinine for non-smoking group.

Anti-inflammatory Activities of Lupane-triterpenoids In Vitro and Their Phytochemical Fingerprinting from Leaves of Acanthopanax gracilistylus

  • Li, Xiao Jun;Dai, Ling;Li, Zhi;Zhang, Xiao Dan;Liu, Xiang Qian;Zou, Qin Peng;Xie, Xia
    • Natural Product Sciences
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    • v.21 no.2
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    • pp.104-110
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    • 2015
  • The activities on the inhibition of NO on LPS-induced RAW 264.7 macrophages were investigated in this work. A simple and sensitive method has been developed and validated for fingerprinting analysis of leaves of Acanthopanax gracilistylus W.W. Smith (AGS). The cytotoxicity and inhibition of NO on LPS-induced RAW 264.7 cells of the extract and triterpenoids were determined. Optimal conditions of HPLC analysis were established as follows. The separation was performed with an ODS-C18 column at $30^{\circ}C$, the detected wavelength was 210 nm, the flow rate was 1 mL/min, and the mobile phase consisted of acetonitrile (0.05% phosphoric acid)-0.05% phosphoric acid solution with gradient elution. Our results showed that impressic acid and acankoreaogenin was more effective on the inhibition of NO than the methanol extract and other compounds. There were seventeen peaks coexisted with similarities above 0.95 and nine lupane-triterpenoids including acankoreaogenin and impressic acid detected and identified. The result of anti-inflammatory activities provides a potential explanation for the use of AGS leaves as a herbal medicine in the treatment of inflammatory diseases. Our results also show that acankoreanogenin and impressic acid may be potentially useful in developing new anti-inflammatory agents. In addition, the fingerprint chromatography clearly illustrated and confirmed the material basis for the anti-inflammatory activities of this plant.

Study on analytical method of residual benzimidazole anthelmintics in meat by LC/MS (LC/MS를 이용한 식육중 잔류 벤지미다졸계 구충제 분석법 연구)

  • Choi Eun-Young;Seo Heyng-Seok;Baek Kui-Jeong;Hur Boo-Hong;Seo Lee-Won;Joung Dong-Suk
    • Korean Journal of Veterinary Service
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    • v.28 no.1
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    • pp.81-89
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    • 2005
  • Recently, mass spectrometry coupled with liquid chromatography (LC/MS) has been a preferred technique for determination of organic compounds in complex matrixes. LC/MS provides a high degree sensitivity and specificity of the compounds of interest. The purpose of this study was to confirm analytical method of residual 6 benzimidazoles (thiabendazole, oxfendazole, mebendazole, albendazole, flubendazole and fenbendazole) in meat by LC/MS. Benzimidazoles were analyzed by LC/MS on XTerra $C_{18}$ column with 0.01% trifluoroacetic acid-acetonitrile (TFA) in a gradient mode as mobile phase, and that were identified by electrospray ionization with selected ion recording mode at 150-350 amu mass range. Residual benzimidazoles were extracted from tissue with ethylacetate, and elute benzimidazoles with $50\%$ acetonitrile. In the LC/MS analysis of benzimidazoles, signal to noise ratio was showed relatively high in the positive mode and special ion in the quality analysis was determined via $[M+H]^+$ and Fragment ions. A spectrum of benzimidazoles was showed from all 6 benzimidazoles

A Study on the Analysis of 89Sr and 90Sr with Cerenkov Radiation and Liquid Scintillation Counting Method (첼렌코프광과 액체섬광계수법을 이용한 89Sr 및 90Sr 분석에 대한 연구)

  • Lee, Myung-Ho;Chung, Geun-Ho;Cho, Young-Hyun;Choi, Geun-Sik;Lee, Chang-Woo
    • Analytical Science and Technology
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    • v.15 no.1
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    • pp.20-25
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    • 2002
  • An accurate and simple analytical technique for $^{89}Sr$ and $^{90}Sr$, overcoming the demerits of the conventional method, has been developed with extraction chromatography and liquid scintillation counting. The Sr fraction was separated from hindrance elements with oxalate coprecipitation or cation exchange resin and purified with Sr-Spec column. With liquid scintillation counter, $^{89}Sr$ was measured by Cerenkov radiation method, and $^{90}Sr$ was measured by spectrum unfolding method. The developed radioactive strontium separation method was validated by application to the IAEA-reference material (IAEA-375, Soil) and radioactive waste samples.