• Title/Summary/Keyword: capillary electrophoresis

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Forensic STR Analysis of Mixed Chimerism after Allogeneic Bone Marrow Transplantation

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.193-196
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    • 2010
  • Multiplex PCR-based short tandem repeat (STR) analysis is considered as a good tool for monitoring bone marrow engraftment after sex-mismatched allogeneic transplantation and provides a sensitive and accurate assessment of the contribution of both donor and/or recipient cells in post-transplantation specimens. Forensic STR analysis and quantitative real time PCR are used to determine the proportion of donor versus recipient each contained within the total DNA. The STR markers were co-amplified in a single reaction by using commercial $PowerPlex^{(R)}$ 16 system and $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ / $Yfiler^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI $PRIS^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The $GeneMapper^{TM}$ ID software were used for size calling and analysis of STR profiles. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA / Y Human Male DNA Quantification Kit The intent of this study was to analyze the ratio of donor versus recipient cells in the post-transplant peripheral blood, spleen, lung and kidney specimens. Specimens were taken from the traffic accident male victim who had been engrafted from bone marrow female donor. Blood and spleen specimens displayed female donor DNA profile. Kidney specimen showed male recipient DNA profile. Interestingly, lung tissue showed mixed profiles. The findings of this study indicate that the forensic STR analysis using fluorescence labeling PCR combined with capillary electrophoresis is quick and reliable enough to assess the ratio of donor versus recipient cells and to monitor the mixed chimeric patterns.

Development of Photo-diode for LOC fluorescence detector (LOC 형광검출 소자를 위한 광 다이오드의 제작 및 특성 평가)

  • Kim, Ju-Hwan;Shin, Kyeong-Sik;Kim, Yong-Kook;Kim, Sang-Sik;Ju, Byeong-Kwon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2003.04a
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    • pp.100-103
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    • 2003
  • Signal detection technologies such as fluorescence, charge and electrochemical detection used in the monolithic capillary electrophoresis system to convert the biochemical reaction into the electrical signal. The fluorescence detection using photodiodes that measure fluorescence emitted from eluting molecules is widely used for the monolithic capillary electrophoresis system. In this paper, in order to fabricate a photosensor has the increased sensitivity, we investigated on the sensitivity of general type and p-i-n type diode. The p-i-n diode has higher sensitivity than photodiode. Considering these results, we fabricated p-i-n diodes on the high resistive$(4k{\Omega}{\cdot}cm)$ wafer into rectangle and finger pattern and compared internal resistance of each pattern. The internal resistance of p-i-n diode can be decreased by the application of finger pattern has parallel resistance structure from $571{\Omega}$ to $393{\Omega}$.

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Determination of Stability Constants of the Inclusion Complexes of ${\beta}$-Blockers in Heptakis (2,3-Dimethyl-6-Sulfato)- ${\beta}$-Cyclodextrin

  • Phuong, Nuyen Thi;Lee, Kyung-Ah;Kim, Kyung-Ho;Choi, Jung-Kap;Kim, Jong-Moon;Kang , Jong-Seong
    • Archives of Pharmacal Research
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    • v.27 no.12
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    • pp.1290-1294
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    • 2004
  • The ${\beta}$-blockers possess at least one chiral center and the S(-)-enantiomer shows higher affinity for binding to the ${\beta}$-adrenergic receptors than antipode. The stability constants of acebutolol, celiprolol, propranolol and terbutaline in the inclusion complexes with single-isomer heptakis (2,3-dimethyl-6-sulfato)- ${\beta}$-cyclodextrin (HDMS-${\beta}$-CD) were determined by capillary electrophoresis. The approximation and linear double reciprocal methods were adapted with comparable results. Among the ${\beta}$-blockers studied, propranolol had the lowest stability constant but the highest enantioselectivity, indicating that the magnitudes of the stability constants carried little information about enantioseparation. The magnitudes of enantioselectivities between the enantiomer pair were in the order of propranolol > celiprolol > terbutaline > acebutolol.

Effect of Coexisting Ions on Electrokinetic Injection in Capillary Electrophoresis Analysis (모세관 전기영동 분석에서 계면 동전기 주입에 미치는 공존 이온의 영향)

  • Lee, Kwang-Woo;Jeon, Ji-Young;Lee, Kwang-Pill
    • Analytical Science and Technology
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    • v.9 no.1
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    • pp.35-42
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    • 1996
  • A rapid analytical method based on capillary electrophoresis is described for the determination of trace anions in high-purity chemicals which is used to prevent corrosion demage in nuclear power plants. Separations are carried out at 20kV using trimethylsilane-coated fused-silica capillary ($70cm{\times}50$ or $75{\mu}m$ i.d.) with the electrolyte of 5mM Chromate(pH=8). Detection was achieved using on-column indirect photometry at 254nm. The simultaneous analysis of inorganic anions, chloride, nitrate, sulfate, azide and phosphate was performed using methods of hydrodynamic(>1ppm) and / or electrokinetic(<1ppm) injection. The results of studies on the coexisting anions on analyte ions shows that peak responses of analyte in hydrodynamic injection is constant without effect of coexisting anions, but those of analysis in electrokinetic injection is strongly dependant upon the kind of coexisting anions and its ionic mobility. The analyte enrichment rate, hence peak response, is positive relationship with the resistance of the sample solution. Thus, appropriate measures, such as standard addition or internal standard technique, must be used to account for differences in conductance of standard and sample solutions.

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Analysis of the Metabolites of 1,2,4-Trimethylbenzene by Capillary Electrophoresis (모세관 전기영동법을 이용한 1,2,4-트리메틸벤젠 대사체의 분석)

  • Kang, Jong-Seong;Hong, Cheong-Hee;Lim, Jeong-Mi;Lee, Yong-Moon;Jang, Jae-Yeon
    • Analytical Science and Technology
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    • v.12 no.4
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    • pp.326-331
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    • 1999
  • The metabolites of 1,2,4-trimethylbenzene (TMB) were synthesized and determined by capillary electrophoresis (CE). The optimum conditions of CE for the separation and determination of 3,4-, 2,4-, 2,5-dimethylbenzoic acid and 3,4-, 2,4-, 2,5-dimethylhippuric acid from the rat urine were as following: the fused silica capillary($75{\mu}m$ i.d. ${\times}$ 36 cm length, 29 cm to detector) was used and kept at $15^{\circ}C$. The applied voltage was 10㎸ and compounds were detected at UV 210 mnm and 254 nm. The running electrolyte was 0.1 M phosphate buffer (pH 7) containing 15 mM of ${\beta}-CD$ and 3% of 2-propanol. The relative amount of the metabolite of 1,2,4-TMB in the rat urine was 56.7% of 3,4-isomer, 30.5% of 2,4-isomer and 12.8% of 2,5-isomer. This method can be applied to the analysis of TMB-metabolites in human urine.

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Chiral Separation of Quinolone Antibacterial Agent by Capillary Electrophoresis (모세관 전기 영동을 이용한 퀴놀린계 항생제의 광학 이성질체 분석)

  • Gang, Dae Cheon;Jo, Seung Il;Jeong, Du Su;Choe, Gyu Seong;Kim, Yong Seong
    • Journal of the Korean Chemical Society
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    • v.46 no.5
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    • pp.412-429
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    • 2002
  • Chiral separation of gemifloxacin, an quinolone antibacterial agent, using (+)-(18-crown-6)-tetracar-boxylic acid $(18C6H_4)$ as a chiral selector was performed by capillary electrophoresis (CE). Direct analysis of quinolone antibacterial agent in body fluid is beneficial in terms of fast analysis time, multicomponent analysis. However, high con-centration of sodium ion in body fluid can prevent gemifloxacin from interacting with $18C6H_4$ since sodium ion has high affinity with $18C6H_4$ due to the strong charge interaction. Ethylenediaminetetraacetic acid (EDTA), as a chelating ligand, was added in the running buffer in order to reduce the interaction between sodium ion and the chiral selector. Increased separation efficiency and reduced migration time were observed while sodium ion exists in the sample solution at the concentration up to 150 mM.

Enantioselective electrophoretic behavior of lipoic acid in single and dual cyclodextrin systems

  • Le, Thi-Anh-Tuyet;Nguyen, Bao-Tan;Phan, Thanh Dung;Kang, Jong-Seong;Kim, Kyeong Ho
    • Analytical Science and Technology
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    • v.34 no.4
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    • pp.143-152
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    • 2021
  • Capillary electrophoresis (CE) is an effective technique to study chiral recognition because it offers flexibility in adjusting vital factors. Currently, various available cyclodextrins (CDs) can be employed for the chiral separation of numerous analytes. Herein, we investigate the enantioseparation behavior of lipoic acid enantiomers in various types of single and dual CD systems through CE. Additionally, several impacted CE parameters were optimized through the systematic investigation based on the design of experiment (DoE) concept for a single system comprising a heptakis (2,3,6-tri-O-methyl)-β-CD and a dual system containing the combination of the single CD with a sulfated-β-CD. Consequently, absolute enantioresolution was obtained within 15 min on a common standard bare fused-silica capillary (64.5/56 cm in total/effective length, 50/365 ㎛ inner/outer diameter), maintained at 15 ℃ and at an applied voltage of 24 kV. The optimal background electrolyte consisted of 6 mM heptakis (2,3,6-tri-O-methyl)-β-CD dissolved in the solution of 58 mM borate buffer at pH 10. Furthermore, the results of apparent binding constant experiments indicated that the S-enantiomer-heptakis (2,3,6-tri-O-methyl)-β-CD complex exhibited a stronger affinity than its R-enantiomer counterpart. The obtained electrophoretic mobility values could be utilized to interpret the resolution achieved at various CD concentrations and the mobility behavior of the complexes elucidated the migration order of the enantiomers in an electropherogram.

Discrimination of Geographical Origin for Astragalus Root (Astragalus membranaceus) by Capillary Electrophoresis and Near-Infrared Spectroscopy (Capillary electrophoresis 및 근적외선분광분석기를 이용한 황기의 원산지 판별)

  • Kim, Eun-Young;Kim, Jung-Hyun;Lee, Nam-Yun;Kim, Soo-Jeong;Rhyu, Mee-Ra
    • Korean Journal of Food Science and Technology
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    • v.35 no.5
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    • pp.818-824
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    • 2003
  • Capillary electrophoresis (CE) and near-infrared spectroscopy (NIRS) were performed to discriminate astragalus roots (Astragalus membranaceus) according to geographical origin (domestic or foreign). Two-hundred-and-four astragalus roots were extracted with 30% methanol in 0.1 M phosphate buffer (pH 2.5) and separated in a uncoated fused-silica $(50\;{\mu}m{\times}27\;cm)$ capillary. Conditions for optimal analysis included: temperature $-45^{\circ}C$, voltage -14 kV, and pressure injection time -8 sec. The optimal separation buffer was 0.1 M phosphate buffer (pH 2.5) containing 40 mM hexane sulfonic acid with 20% 2-methoxy ethanol. Raw NIR spectra were obtained using NIRS, and modified partial least square regression was used to develop the prediction model. The correlation coefficient and standard error of prediction were 0.915 and 14.3%, respectively. Under the optimal conditions established for CE and NIRS, the geographical origins of the astragalus roots were correctly identified in 80 and 97%, respectively. Astragalus roots that were not discriminated by NIRS were correctly discriminated by CE. Hence, CE and NIRS are potential methods for discriminating the geographical origins of astragalus roots that complement one another.

Fast Microchip Electrophoresis Using Field Strength Gradients for Single Nucleotide Polymorphism Identification of Cattle Breeds

  • Oh, Doo-Ri;Cheong, Il-Cheong;Lee, Hee-Gu;Eo, Seong-Kug;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • v.31 no.7
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    • pp.1902-1906
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    • 2010
  • A microchip electrophoresis (ME) method was developed using a programmed field strength gradients (PFSG) for the single nucleotide polymorphism (SNP) based fast identification of cattle breeds. Four different Korean cattle (Hanwoo) and Holstein SNP markers amplified by allele-specific polymerase chain reaction were separated in a glass microchip filled with 0.5% poly(ethyleneoxide) ($M_r$ = 8 000 000) by PFSG as follows: 750 V/cm for 0 - 14 s, 166.7 V/cm for 14 - 31 s, 83.3 V/cm for 31 - 46 s, and 750 V/cm for 46 - 100 s. The cattle breeds were clearly distinguished within 45 s. The ME-PFSG method was 7 times and 5 times faster than the constant electric field ME method and the capillary electrophoresis- PFSG method, respectively, with a high resolving power ($R_s$ = 5.05 - 9.98). The proposed methodology could be a powerful tool for the fast and simultaneous determination of SNP markers for various cattle breeds with high accuracy.

Capillary Electrophoretic Method for the Determination of (+)-Catechin, (-)-Epicatechin in Grape Seed Ethanol Extract (포도종실 에탄올 추출물에 함유된 (+)-카테킨, (-)-에피카테킨의 모세관 전기영동법에 의한 분석)

  • Choi, One-Kyun;Chung, Yang-Seop;Chung, Ha-Yull
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.513-518
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    • 2005
  • Capillary electrophoresis (CE) method was developed to determine (+)-catechin and (-)-epicatechin contents in grape seed ethanol extract. CE separation was achieved using 100 mM phosphate and borate buffer at pH 6.0 as background electrolyte and fused silica capillary with 50 microns x 375 microns O.D. (effective length 20.0cm) maintained at $25^{\circ}C$. The applied voltage was 10kV, and detection was performed by DAD at 210 nm, Two catechins were well separated within 6 min with repeatability of <0.8% RSD for migration time and <2.0% RSD for peak area, and correlation coefficients higher than 0.994 were obtained from 58.0 to 174.0 mg/L with detection limit of 0.035 mg/L. Separated compounds were successfully determined. CE method was easy to handle and showed good reproducibility. CE method was compared with conventional coloring and HPLC methods, and main advantages of CE method were low amount of sample required, simple pre-sample treatment, good recovery rate, and short analysis time.