• Title/Summary/Keyword: Ion-chromatography

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Mutation Analysis in β2-Adrenergic Receptor Gene by Denaturing High Performance Liquid Chromatography (DHPLC) (DHPLC를 이용한 β2-교감신경수용체 유전자에서의 돌연변이 분석)

  • Park, Sang-Bum;Oh, Chung-Hun;Kim, Jong-Wan;Jang, Won-Cheoul
    • Analytical Science and Technology
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    • v.15 no.3
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    • pp.190-195
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    • 2002
  • We analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed-phase high performance liquid chromatography (IP-RP-HPLC). We extracted genomic DNA from 50 asthma patients, amplified DNA using PCR, and analysed PCR product by DHPLC. As a result, we obtained that mutation frequency was 15 (30%) among 50 cases. Consequently DHPLC mutation detection was confirmed that the result of direct sequencing was coincide exactly.

Purification of Bacillus sp. β-Mannanase and Separation of Xanthan Gum Hydrolysate by Chromatography Methods (Bacillus sp. 유래 β-Mannanase의 정제 및 Chromatography에 의한 Xanthan Gum 가수분해물의 분리)

  • 박귀근
    • 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.

Gas Chromatography-High Resolution Tandem Mass Spectrometry Using a GC-APPI-LIT Orbitrap for Complex Volatile Compounds Analysis

  • Lee, Young-Jin;Smith, Erica A.;Jun, Ji-Hyun
    • Mass Spectrometry Letters
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    • v.3 no.2
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    • pp.29-38
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    • 2012
  • A new approach of volatile compounds analysis is proposed using a linear ion trap Orbitrap mass spectrometer coupled with gas chromatography through an atmospheric pressure photoionization interface. In the proposed GC-HRMS/MS approach, direct chemical composition analysis is made for the precursor ions in high resolution MS spectra and the structural identifications were made through the database search of high quality MS/MS spectra. Successful analysis of a complex perfume sample was demonstrated and compared with GC-EI-Q and GC-EI-TOF. The current approach is complementary to conventional GC-EI-MS analysis and can identify low abundance co-eluting compounds. Toluene co-sprayed as a dopant through API probe significantly enhanced ionization of certain compounds and reduced oxidation during the ionization.

Novel Purification Method of Kv 4.2 Potassium Channel from Rat Brain Membrane

  • Park, Sung-Soo
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.96-103
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    • 2012
  • Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.

High Performance Liquid Chromatography (HPLC) Detection of Malonaldehydethiobarbituric Acid (MA-TBA) Complex in Ground Pork

  • Whang, Key
    • Preventive Nutrition and Food Science
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    • v.4 no.3
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    • pp.171-174
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    • 1999
  • For monitoring lipid oxidation development in cooked ground pork during refrigerationm, malonaldehydethiobarbituric acid(MA-TBA) contents were measured using high performance liquid chromatography(HPLC). As the oxidation proceeded during refergeration, TBA-reaction substances(TBARS) absorbances increased and the corresponding HPLC peak areas also increased proportationately. The correlation coefficient between the HPLC peak areas and MA-TBA absorbance were 0.9979. The treatemtn of cetrimide, an ion pairing agent, gave a complete resolution of the MA-TBA complex and the butanol extraction of the complex increased its recovery by 37.8%. Both cetrimide treatment and butanol extraction are essential steps for analyzing MA-TBA complex in ground pork wiht HPLC. A reliable and specific measurement of NA-TBA in ground pork was successfully performed using HPLC.

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Determination of Iodide in Sulfuric Acid Aqueous Solution by the Ion Chromatography with UV Detection (이온 크로마토그래피와 자외선 검출을 이용한 황산수용액 중의 요오드 음이온 정량)

  • Park, Yang-Soon
    • Analytical Science and Technology
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    • v.14 no.4
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    • pp.306-310
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    • 2001
  • Ion chromatography was applied to determine iodide remained in sulfuric acid aqueous solution after adsorption procedure. Iodide was determined in 0.25 M, 0.5 M and 1 M sulfuric acid solution on time variation. Because sulfuric acid in solution plays as an oxidizer, the concentration of iodide decreased with increasing concentration or sulfuric acid. Thereafter, sulfuric acid was neutralized with sodium hydroxide. Good linearity(r=0.99999) was obtained at the range of 0-20 mg/L 1 in 0.5 M sodium sulfate matrix.

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Development of Conductivity Cell and Suppressor for Capillary Column Ion Chromatography (모세관 컬럼 이온 크로마토그래피를 위한 Conductivity Cell과 Suppressor의 개발)

  • Pyo, Dongjin;Kim, Hohyun
    • Analytical Science and Technology
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    • v.12 no.2
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    • pp.89-93
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    • 1999
  • In this study, conductivity cell and suppressor for micro-column ion chromatography were developed to analyze ions in small columns of samples. With a capillary column, the flow rate of the mobile phase is so small (usually $5{\sim}20{\mu}L/min$) that the usual conductivity cell can not be used. Therefore, we developed a new type of conductivity cell and suppressor which have small inner volumes. The conductivity cell was made with two Pt hypodermic needles (i.d. 0.010 mm) which are slightly separated (about $2{\mu}m$), and the suppressor was made of Nafion tubings. When several anions(fluoride, nitrite, nitrate, chlorate) were analyzed using developed conductivity cell and suppressor, a good chromatogram was obtained.

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Determination of Cyanogenic Compounds in Edible Plants by Ion Chromatography

  • Cho, Hye-Jeon;Do, Byung-Kyung;Shim, Soon-Mi;Kwon, Hoonjeong;Lee, Dong-Ha;Nah, Ahn-Hee;Choi, Youn-Ju;Lee, Sook-Yeon
    • Toxicological Research
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    • v.29 no.2
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    • pp.143-147
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    • 2013
  • Cyanogenic glycosides are HCN-producing phytotoxins; HCN is a powerful and a rapidly acting poison. It is not difficult to find plants containing these compounds in the food supply and/or in medicinal herb collections. The objective of this study was to investigate the distribution of total cyanide in nine genera (Dolichos, Ginkgo, Hordeum, Linum, Phaseolus, Prunus, Phyllostachys, Phytolacca, and Portulaca) of edible plants and the effect of the processing on cyanide concentration. Total cyanide content was measured by ion chromatography following acid hydrolysis and distillation. Kernels of Prunus genus are used medicinally, but they possess the highest level of total cyanide of up to 2259.81 $CN^-$/g dry weight. Trace amounts of cyanogenic compounds were detected in foodstuffs such as mungbeans and bamboo shoots. Currently, except for the WHO guideline for cassava, there is no global standard for the allowed amount of cyanogenic compounds in foodstuffs. However, our data emphasize the need for the guidelines if plants containing cyanogenic glycosidesare to be developed as dietary supplements.

Determination of 11 Phenolic Endocrine Disruptors using Gas Chromatography/Mass Spectrometry-Selected Ion Monitoring in Five Selected Wastewater Influents

  • Kim, Hyub
    • Environmental Engineering Research
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    • v.13 no.4
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    • pp.216-223
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    • 2008
  • An efficient method for the simultaneous determination of eleven phenolic endocrine-disrupting chemicals (EDCs) present in wastewater influent samples was described. The 11 phenolic EDCs including alkylphenols, chlorophenols, and bisphenol A were determined by gas chromatography/mass spectrometry-selected ion monitoring (GC/MS-SIM) following two work-up methods for comparison; isobutoxycarbonyl (isoBOC) derivatization and tert-butyldimethylsilyl (TBDMS) derivatization. The wastewater influent samples containing the 11 EDCs were adjusted to pH 2 with $H_2SO_4$ and then cleaned up with n-hexane. Next, they were subjected to solid-phase extraction (SPE) with XAD-4 resin and subsequently converted to isoBOC or TBDMS derivatives for sensitivity analysis with gas chromatography/mass spectrometry-selected ion monitoring (GC/MSSIM). Following isoBOC derivatization and TBDMS derivatization, the recoveries were 86.6-105.2% and 97.6-142.7%, the limits of quantitation (LOQ) for the 11 phenolic EDCs for SIM was 0.001-0.050 ng/mL and 0.003-0.050 ng/mL, and the SIM responses were linear with the correlation coefficient varying by 0.9717-0.9995 and 0.9842-0.9980, respectively. When these methods were applied to five selected wastewater influent samples, for isoBOC derivatization and TBDMS derivatization the ranges of concentration detected were 0.2-99.6 ng/mL and 0.4-147.4 ng/mL, respectively.

Determination of Clotiazepam in the Plasma Using Gas Chromatography/Mass Spectrometry with an Ion-Trap Detector and its Application to Pharmacokinetics in Healthy Volunteers

  • Kwon, Oh-Seung;Kim, Seung-Yong;Chung, Youn-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.2
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    • pp.123-129
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    • 2006
  • A method determining the plasma concentration of clotiazepam was developed by using gas chromatography/mass spectrometry with an ion-trap detector and was validated for applying pharmacokinetics to human volunteers orally taken 5 mg dose of clotiazepam. The detection limit was 1 ng/ml and the limit of quantitation was 5 ng/mt. Intraday reproducibility and accuracy bias % were less than 8.2 and 10.2% with inter-day variations for those being within 7.0 and 13.8%, respectively. The recovery of clotiazepam was higher than 87%. The principal pharmacokinetic parameters were determined from the plasma concentration-time plot by non-compartmental or two-compartmental analysis. In non-compartmental analysis, the elimination half-life of 10.4 hr and the area under the curve of 651.3 ng hr/ml were determined, and the maximal concentration (158.6 ng/ml) in the plasma was obtained at 0.56 hr post-dose. The developed method can be appropriate to apply pharmacokinetics and bioequivalence of clotiazepam.