• 제목/요약/키워드: Capillary Electrophoresis

검색결과 175건 처리시간 0.027초

IPA 저온 접합법을 이용한 PMMA Micro CE Chip의 제작 (Fabrication of PMMA Micro CE Chip Using IPA Assisted Low-temperature Bonding)

  • 차남구;박창화;임현우;조민수;박진구
    • 한국재료학회지
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    • 제16권2호
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    • pp.99-105
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    • 2006
  • This paper reports an improved bonding method using the IPA (isopropyl alcohol) assisted low-temperature bonding process for the PMMA (polymethylmethacrylate) micro CE (capillary electrophoresis) chip. There is a problem about channel deformations during the conventional processes such as thermal bonding and solvent bonding methods. The bonding test using an IPA showed good results without channel deformations over 4 inch PMMA wafer at $60^{\circ}C$ and 1.3 bar for 10 minutes. The mechanism of IPA bonding was attributed to the formation of a small amount of vaporized acetone made from the oxidized IPA which allows to solvent bonding. To verify the usefulness of the IPA assisted low-temperature bonding process, the PMMA micro CE chip which had a $45{\mu}m$ channel height was fabricated by hot embossing process. A functional test of the fabricated CE chip was demonstrated by the separation of fluorescein and dichlorofluorescein. Any leakage of liquids was not observed during the test and the electropherogram result was successfully achieved. An IPA assisted low-temperature bonding process could be an easy and effective way to fabricate the PMMA micro CE chip and would help to increase the yield.

Simultaneous Detection of 10 Foodborne Pathogens using Capillary Electrophoresis-Based Single Strand Conformation Polymorphism

  • Oh, Mi-Hwa;Hwang, Hee-Sung;Chung, Bo-Ram;Paik, Hyun-Dong;Han, Sang-Ha;Kang, Sun-Moon;Ham, Jun-Sang;Kim, Hyoun-Wook;Seol, Kuk-Hwan;Jang, Ae-Ra;Jung, Gyoo-Yeol
    • 한국축산식품학회지
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    • 제32권2호
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    • pp.241-246
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    • 2012
  • This report outlines the development of a rapid, simple, and sensitive detection system for pathogenic bacteria using a capillary electrophoresis-based, single strand conformation polymorphism (CE-SSCP) combined with PCR. We demonstrate that this method, used with primers targeting the V4 region of the16S rRNA gene, is capable of the simultaneous detection of 10 microbes that could be associated with foodborne illness, caused by animal-derived foods: Salmonella enterica, Listeria monocytogenes, Escherichia coli O157:H7, Campylobacter jejuni, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Yersinia enterocolitica, Vibrio parahaemolyticus, and Enterobacter sakazakii. The traditional detection techniques are time-consuming and labor-intensive, due to the necessary task of separate cultivation of each target species. As such, the CE-SSCP-PCR method, that we have developed, has the potential to diagnose pathogens rapidly, unlike the traditional technique, in order to prevent foodborne illness in a much more efficient manner.

모세관 젤 전기영동을 이용한 안드로젠 수용체 유전자 분석 (Analysis of Androgen Receptor Gene by Capillary Gel Eelectrophoresis)

  • 김용성;백승권;강충무;강대천;이수만;최병옥;정기화;최규성
    • 대한화학회지
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    • 제50권1호
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    • pp.37-45
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    • 2006
  • 연구에서는 모세관 젤 전기 영동에서 분자체 역할을 하는 poly(ethylene oxide)를 이용하여 남성불임(Androgen receptor) 유전자의 정확한 CAG 반복 배열의 길이를 측정하였다. 표준물질인 X174 RF/Hae III, 100bp step ladder를 이용하여 non-linear regression analysis를 통한 81명의 한국의 불임 남성과 89명의 정상적인 한국 남성을 대조군으로 하여, 남성 불임과 관련된 AR gene의 exon 1의 CAG 반복 수를 측정하였다. 불임 남성의 CAG 반복 길이는 24.97±2.6 range, 17-29)이고, 정상인 남성은 23.99±2.4(range, 18-29)이다. P값은 0.018로써(<0.05) 두 그룹의 결과는 통계학적으로 주목 할 수 있는 차이를 나타내었다.

Metabolomic approach to key metabolites characterizing postmortem aged loin muscle of Japanese Black (Wagyu) cattle

  • Muroya, Susumu;Oe, Mika;Ojima, Koichi;Watanabe, Akira
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권8호
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    • pp.1172-1185
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    • 2019
  • Objective: Meat quality attributes in postmortem muscle tissues depend on skeletal muscle metabolites. The objective of this study was to determine the key metabolic compounds and pathways that are associated with postmortem aging and beef quality in Japanese Black cattle (JB; a Japanese Wagyu breed with highly marbled beef). Methods: Lean portions of Longissimus thoracis (LT: loin) muscle in 3 JB steers were collected at 0, 1, and 14 days after slaughter. The metabolomic profiles of the samples were analyzed by capillary electrophoresis time-of-flight mass spectrometry, followed by statistical and multivariate analyses with bioinformatics resources. Results: Among the total 171 annotated compounds, the contents of gluconic acid, gluconolactone, spermidine, and the nutritionally vital substances (choline, thiamine, and nicotinamide) were elevated through the course of postmortem aging. The contents of glycolytic compounds increased along with the generation of lactic acid as the beef aging progressed. Moreover, the contents of several dipeptides and 16 amino acids, including glutamate and aromatic and branched-chain amino acids, were elevated over time, suggesting postmortem protein degradation in the muscle. Adenosine triphosphate degradation also progressed, resulting in the generation of inosine, xanthine, and hypoxanthine via the temporal increase in inosine 5'-monophosphate. Cysteine-glutathione disulfide, thiamine, and choline increased over time during the postmortem muscle aging. In the Kyoto encyclopedia of genes and genomes database, a bioinformatics resource, the postmortem metabolomic changes in LT muscle were characterized as pathways mainly related to protein digestion, glycolysis, citric acid cycle, pyruvate metabolism, pentose phosphate metabolism, nicotinamide metabolism, glycerophospholipid metabolism, purine metabolism, and glutathione metabolism. Conclusion: The compounds accumulating in aged beef were shown to be nutritionally vital substances and flavor components, as well as potential useful biomarkers of aging. The present metabolomic data during postmortem aging contribute to further understanding of the beef quality of JB and other breeds.

Metabolomic profiling of postmortem aged muscle in Japanese Brown beef cattle revealed an interbreed difference from Japanese Black beef

  • Susumu Muroya;Riko Nomura;Hirotaka Nagai;Koichi Ojima;Kazutsugu Matsukawa
    • Animal Bioscience
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    • 제36권3호
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    • pp.506-520
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    • 2023
  • Objective: Japanese Brown (JBR) cattle, especially the Kochi (Tosa) pedigree (JBRT), is a local breed of moderately marbled beef. Despite the increasing demand, the interbreed differences in muscle metabolites from the highly marbled Japanese Black (JBL) beef remain poorly understood. We aimed to determine flavor-related metabolites and postmortem metabolisms characteristic to JBRT beef in comparison with JBL beef. Methods: Lean portions of the longissimus thoracis (loin) muscle from four JBRT cattle were collected at 0, 1, and 14 d postmortem. The muscle metabolomic profiles were analyzed using capillary electrophoresis time-of-flight mass spectrometry. The difference in post-mortem metabolisms and aged muscle metabolites were analyzed by statistical and bioinformatic analyses between JBRT (n = 12) and JBL cattle (n = 6). Results: A total of 240 metabolite annotations were obtained from the detected signals of the JBRT muscle samples. Principal component analysis separated the beef samples into three different aging point groups. According to metabolite set enrichment analysis, post-mortem metabolic changes were associated with the metabolism of pyrimidine, nicotinate and nicotinamide, purine, pyruvate, thiamine, amino sugar, and fatty acid; citric acid cycle; and pentose phosphate pathway as well as various amino acids and mitochondrial fatty acid metabolism. The aged JBRT beef showed higher ultimate pH and lower lactate content than aged JBL beef, suggesting the lower glycolytic activity in postmortem JBRT muscle. JBRT beef was distinguished from JBL beef by significantly different compounds, including choline, amino acids, uridine monophosphate, inosine 5'-monophosphate, fructose 1,6-diphosphate, and betaine, suggesting interbreed differences in the accumulation of nucleotide monophosphate, glutathione metabolism, and phospholipid metabolism. Conclusion: Glycolysis, purine metabolism, fatty acid catabolism, and protein degradation were the most common pathways in beef during postmortem aging. The differentially expressed metabolites and the relevant metabolisms in JBRT beef may contribute to the development of a characteristic flavor.

Chiral Purity Test of Bevantolol by Capillaryelectrophoresis and High Performance Liquid Chromatography

  • Long, Pham Hai;Trung, Tran Quoc;Oh, Joung-Won;Kim, Kyeong-Ho
    • Archives of Pharmacal Research
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    • 제29권9호
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    • pp.808-813
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    • 2006
  • Two methods for the chiral purity determination of bevantolol were developed, namely capillary electrophoresis (CE) using carboxymethyl-${\beta}$-cyclodextrin (CM-${\beta}$-CD) as a chiral selector and high-perfomance liquid chromatography (HPLC) using a chiral stationary phase. In the HPLC method, the separation of bevantolol enantiomers was performed on a Chiralpak AD-H column by isocratic elution with n-hexane-ethanol-diethylamine (10:90:0.1, v/v/v) as mobile phase. In the CE method, bevantolol enantiomers were separated on an uncoated fused silica capillary with 50 mM amonium phosphate dibasic adjusted to a pH 6.5 with phosphoric acid containing 15 mM CM-${\beta}$-CD as running buffer. Validation data such as linearity, recovery, detection limit, and precision of the two methods are presented. The detection limits of S-(-)-bevantolol were 0.1% and 0.05% for CE and HPLC method, respectively and R-(+)-bevantolol were 0.15% and 0.05% for CE and HPLC method, respectively. There was generally good agreement between the HPLC and CE results.

NEAR INFRARED TRANSFLECTANCE SPECTROSCOPY (NIRS) IN PHYTOCHEMISTRY

  • Huck, C.W.;W.Guggenbichler;Bonn, G.K.
    • 한국근적외분광분석학회:학술대회논문집
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    • 한국근적외분광분석학회 2001년도 NIR-2001
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    • pp.3114-3114
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    • 2001
  • During the last years phytochemistry and phytopharmaceutical applications have developed rapidly and so there exists a high demand for faster and more efficient analysis techniques. Therefore we have established a near infrared transflectance spectroscopy (NIRS) method that allows a qualitative and quantitative determination of new polyphenolic pharmacological active leading compounds within a few seconds. As the NIR spectrometer has to be calibrated the compound of interest has at first to be characterized by using one or other a combination of chromatographic or electrophoretic separation techniques such as thin layer chromatography (TLC), high performance liquid chromatography (HPLC), capillary electrophoresis (CE), gas chromatography (GC) and capillary electrochromatography (CEC). Both structural elucidation and quantitative analysis of the phenolic compound is possible by direct coupling of the mentioned separation methods with a mass spectrometer (GC-MS, LC-MS/MS, CE-MS, CEC-MS) and a NMR spectrometer (LC-NMR). Furthermore the compound has to be isolated (NPLC, MPLC, prep. TLC, prep. HPLC) and its structure elucidated by spectroscopic techniques (UV, IR, HR-MS, NMR) and chemical synthesis. After that HPLC can be used to provide the reference data for the calibration step of the near infrared spectrometer. The NIRS calibration step is time consuming, which is compensated by short analysis times. After validation of the established NIRS method it is possible to determine the polyphenolic compound within seconds which allows to raise the efficiency in quality control and to reduce costs especially in the phytopharmaceutical industry.

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모세관 전기이동법에 의한 생약제제중 베르베린, 계피산 및 글리시리진의 동시 정량 (Simultaneous Determination of Berberine, Cinnamic Acid and Glycyrrhizin in Pharmaceutical Formulations by Capillary Electrophoresis with Diode-Array Detection)

  • 강성호;정화진;윤형중;정두수
    • 대한화학회지
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    • 제41권2호
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    • pp.98-104
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    • 1997
  • 한국에서 전통적으로 사용되고 있는 생약제제 중 베르베린, 계피산 및 글리시리진의 정량을 위한 간단하고, 정확하고, 재현성 있는 모세관 전기이동법을 개발하였다. 이 성분들의 분리는 25$^{\circ}C$, 20 mM 인산완충액(pH 7.5)에서 실리카 캐필러리($57 cm{\times}75 {\mu}m$ i.d.)를 사용하여 수행하였다. 350 V/cm의 전기장으로 베르베린, 계피산 및 글리시리진의 동시 정량을 13분내에 할 수 있었다. 검정곡선은 베르베린에 대해서는 1~100 ${\mu}g/mL$, 계피산은 0.3~100 ${\mu}g/mL$, 글리시리진은 2.5~100 ${\mu}g/mL$의 농도 범위 안에서 좋은 직선성을 보여 주었다. 이 성분들에 대한 상대 표준편차(n=5)의 범위는 0.96∼2.35%이었다. 베르베린, 계피산 및 글리시리진에 대한 검출한계(S/N=3)는 각각 0.5, 0.1 및 2.0 ${\mu}g/mL$이었다. 이론단수는 181,000(베르베린), 88,000(계피산) 및 169,000(글리시리진)이었다. HPLC에서는 3,100~4,800이었다.

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Microchip상에서 효율적인 DNA 분석을 위한 반복단위 단백질의 생산 (Production of Repetitive Polypeptides for an Efficient DNA Analysis on a Microchip)

  • 이현진;최석진;서태석;원종인
    • KSBB Journal
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    • 제25권2호
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    • pp.199-204
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    • 2010
  • Drag-tag으로 사용될 반복단위 단백질을 생물학적인 방법을 통해 생산함으로써 수용액 내에서 DNA 분리가 가능함을 확인하였다. 서로 다른 크기를 갖는 두 종류의 반복단위 단백질을 디자인하였고, 이를 발현시킨 뒤 정제하였다. 정제된 반복단위 단백질에 형광 dye를 포함하고 있는 100 base의 DNA를 연결하였고, 이 연결 물질을 모세관 내부가 수용액으로 충진된 microchip 상에서 전기영동 하였다. 그 결과 생물학적으로 생산된 반복단위 단백질이 SNP 분석과 같은 빠르고 효율적인 DNA 분석에 적합한 후보물질로 사용될 수 있음을 확인하였다.

아조염료와 디아조 성분의 분리 및 비교에 관한 연구 (Study for the separation and comparison of azo dyes and their diazo components)

  • 정혁
    • 분석과학
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    • 제19권1호
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    • pp.50-57
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    • 2006
  • 계면활성제를 완충용액에 섞어서 사용하는 미셀 모세관 전기영동법(micellar electrokinetic capillary chromatography, MECC)을 이용하여, 아조염료의 합성성분이면서 동시에 아조염료가 분해될 때 생성되는 H-acid modifier 혹은 2-naphthylamine-1,5-disulfonic acid 등의 디아조 성분에 대한 분석을 수행하고, 이 분리 결과를 Ion-Paring 메카니즘을 이용한 고성능 액체 크로마토그래피법과 비교하였다. 이 방법을 이용하여 Direct Blue 2, Direct Blue 6, Direct Blue 15 등의 직접염료 그리고 Reactive Orange 4와 같은 반응성 염료의 혼합용액을 완전히 분리할 수 있었다. 특히 각 염료의 환원용액을 H-acid, J-acid, ${\gamma}$-acid, orthanilic acid, sulfanilic acid 2-naphthylamine-1,5-disulfonic acid 등의 표준물질과 비교 분석한 결과, 사용한 각 염료의 디아조 성분인 H-acid modifier 혹은 2-naphthylamine-1,5-disulfonic acid에 대한 정보를 얻을 수 있었다. 본 연구의 결과를 토대로 Ion-Pair 크로마토그래피법과 모세관 전기영동법은 미지의 염료에 대한 성분확인 및 디아조 혹은 커플링 성분분석에 응용할 수 있음을 알 수 있었다.