• Title/Summary/Keyword: UV and fluorescence detection

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Applications of Capillary Electrophoresis and Microchip Capillary Electrophoresis for Detection of Genetically Modified Organisms

  • Guo, Longhua;Qiu, Bin;Xiao, Xueyang;Chen, Guonan
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.823-832
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    • 2009
  • In recent years, special concerns have been raised about the safety assessment of foods and food ingredients derived from genetically modified organisms (GMOs). A growing number of countries establish regulations and laws for GMOs in order to allow consumers an informed choice. In this case, a lot of methods have been developed for the detection of GMOs. However, the reproducibility among methods and laboratories is still a problem. Consequently, it is still in great demand for more effective methods. In comparison with the gel electrophoresis, the capillary electrophoresis (CE) technology has some unique advantages, such as high resolution efficiency and less time consumption. Therefore, some CE-based methods have been developed for the detection of GMOs in recent years. All kinds of CE detection methods, such as ultraviolet (UV), laser induced fluorescence (LIF), and chemiluminescence (CL) detection, have been used for GMOs detection. Microchip capillary electrophoresis (MCE) methods have also been used for GMOs detection and they have shown some unique advantages.

Application of Quantum-dot Nanocrystals for Cyanobacterial Toxin-Microcystin Detection (나노크리스탈 Quantum-dot을 적용한 남조류 독소 Microcystin 탐지 연구)

  • Lee, Jinwook;Yu, Hye-Weon;Kim, In S.
    • Journal of Korean Society on Water Environment
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    • v.23 no.5
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    • pp.705-711
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    • 2007
  • Green quantum-dot nanocrystal (QD525) with anti-microcystin monoclonal antibody was applied for detection of microcystin, a monocyclic peptide hepatotoxin, extracted from the culture of Microcystis aeruginosa. The presence of microcystin in the cell lysate was verified by HPLC analysis with UV absorbance at 238 nm. Microcystis cell extract exhibited fluorescence emission spectra, which peak was around 460 nm because of their complex organic substances. When a spherical QD525 antibody conjugates (10~20 nm in diameter) were bound to the microcystins in the Microcystis cell lysate, the fluorescence intensity of the primary peak at 525 nm diminished while the secondary emission peak at 460 nm slightly increased intensities. It is due to energy transfer from the primary (major) to the secondary (minor) peak, resulting from physical deformation of QD525 and different environmental factors. On the other hand, other cell extracts did not show any fluorescence emission change. This study is very available for detecting and monitoring the microcystin because it is one step assay without washing step and portable spectrophotometer makes on-site measurement possible. For health risk assessment of the microcystin, the reliable and rapid system to detect and quantify microcystin is seriously required.

"Turn-on" type colorimetric/fluorimetric probe for selective detection of Cu2+ at neutral pH condition

  • Lee, Hyun Jung;Saleem, Muhammad;Lee, Ki Hwan
    • Rapid Communication in Photoscience
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    • v.4 no.4
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    • pp.88-90
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    • 2015
  • The design and development of fluorescent chemosensors have recently been intensively explored for sensitive and specific detection of environmentally and biologically relevant metal ions in aqueous solution and living cells. Herein, we report the photophysical results of rhodamine B based fluorogenic and chromogenic receptor for selective copper detection in the complete organic or mixed aqueous-organic media at neutral pH under ambient condition. The ligand exhibited the remarkable increment in the fluorescence emission and UV-visible absorption signal intensities at 587 and 547 nm, respectively, on induction of copper ion while the ligand solution remain completely silent on addition of varieties of other metal ions.

Determination of Catechin Compounds in Korean Green Tea Infusions under Various Extraction Conditions by High Performance Liquid Chromatography

  • Cheong, Won-Jo;Park, Moon-Hee;Kang, Gyoung-Won;Ko, Joung-Ho;Seo, You-Jin
    • Bulletin of the Korean Chemical Society
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    • v.26 no.5
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    • pp.747-754
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    • 2005
  • Liquid chromatographic methods with UV and fluorescence detection have been used to determine the levels of (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, and (-)-epigallocatechin gallate in Korean green tea infusions. The extracts of Korean green tea leaves or powders in water at various temperatures (100 ${^{\circ}C}$, 80 ${^{\circ}C}$, 60 ${^{\circ}C}$) and time, were washed with chloroform and re-extracted to ethyl acetate. The ethyl acetate phase was dried and re-dissolved in methanol and analyzed. Five catechin compounds were separated by gradient elution. The flavonoids were found decomposed on prolonged extraction, thus exhaustive extraction by a Soxhlet apparatus was found useless for green tea. Some unknown components were found in the extracts at 100 ${^{\circ}C}$. When the green tea was filtered and re-extracted with new fresh water, still some flavonoids were extracted. However, the contents of flavonoids in the third extract were found negligible. The flavonoid extraction rate of green tea powders was higher than that of green tea leaves, but flavonoid decomposition of green tea powders was also faster than that of green tea leaves. The traditional way of drinking green tea was found appropriate in view of flavonoids intake.

Complex Detection Between Transcription Regulator and Promoter DNA by UV Spectroscopic Method

  • Lee, Kyungmin;Gang, Jongback
    • Journal of Integrative Natural Science
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    • v.5 no.3
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    • pp.163-167
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    • 2012
  • UV spectrophotometer was used to detect protein-DNA complex from DNA melting profile under constant temperature increase. Melting temperature (Tm) was $43^{\circ}C$ in copA duplex DNA alone. In the presence of Proteus mirabilis transcription regulator protein (PMTR) protein at 0.2 and 0.4 ${\mu}M$, Tm's were $45{\pm}0.5$ and $47.6{\pm}0.6^{\circ}C$, respectively. According to fluorescence polarization and gel shift assay. PMTR:copA complex was detected by the retarded migration on gel and the dissociation constant ($K_d$) was $(9.2{\pm}2.8){\times}10^{-9}M$.

Optimal Optical Filters of Fluorescence Excitation and Emission for Poultry Fecal Detection

  • Kim, Tae-Min;Lee, Hoon-Soo;Kim, Moon-S.;Lee, Wang-Hee;Cho, Byoung-Kwan
    • Journal of Biosystems Engineering
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    • v.37 no.4
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    • pp.265-270
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    • 2012
  • Purpose: An analytic method to design excitation and emission filters of a multispectral fluorescence imaging system is proposed and was demonstrated in an application to poultry fecal inspection Methods: A mathematical model of a multispectral imaging system is proposed and its system parameters, such as excitation and emission filters, were optimally determined by linear discriminant analysis (LDA). An alternating scheme was proposed for numerical implementation. Fluorescence characteristics of organic materials and feces of poultry carcasses are analyzed by LDA to design the optimal excitation and emission filters for poultry fecal inspection. Results: The most appropriate excitation filter was UV-A (about 360 nm) and blue light source (about 460 nm) and band-pass filter was 660-670 nm. The classification accuracy and false positive are 98.4% and 2.5%, respectively. Conclusions: The proposed method is applicable to other agricultural products which are distinguishable by their spectral properties.

Microfluidic Fabrication of Conjugated Polymer Sensor Fibers (미세유동을 이용한 공액 고분자 센서 섬유 제작)

  • Yoo, Imsung;Song, Simon
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.38 no.10
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    • pp.853-858
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    • 2014
  • We propose a fabrication method for polydiacetylene (PDA)-embedded hydrogel microfibers on a microfluidic chip. These fibers can be applied to the detection of cyclodextrines (CDs), which are a family of sugar and aluminum ions. PDA, a family of conjugated polymers, has unique characteristics when used for a sensor, because it undergoes a blue-to-red color transition and nonfluorescence-to-fluorescence transition in response to environmental stimulation. PDAs have different sensing characteristics depending on the head group of PCDA. By taking advantage of ionic crosslinking-induced hydrogel formation and the 3D hydrodynamic focusing effect on a microfluidic chip, PCDA-EDEA-derived diacetylene (DA) monomer-embedded microfibers were successfully fabricated. UV irradiation of the fibers afforded blue-colored PDA, and the resulting blue PDA fibers underwent a phase transition to red and emitted red fluorescence upon exposure to CDs and aluminum ions. Their fluorescence intensity varied depending on the CDs and aluminum ion concentrations. This phase transition was also observed when the fibers were dried.

The Application of a Laser to the Chemical Characterization of Radionuclides

  • Park, Y.J.;Park, K.K.;M/Y. Suh;S.K. Yoon;Park, Y.S.;Kim, D.Y.;Kim, W.H.
    • Nuclear Engineering and Technology
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    • v.32 no.5
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    • pp.446-456
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    • 2000
  • Laser induced photoacoustic, fluorescence, and photon correlation spectroscopies were applied to the chemical characterization of radionuclides in connection with the radiowaste treatment and disposal. Their measuring principles and systems were briefly described together with their advantages over conventional spectroscopies. Also, other applications of lasers are introduced. Laser induced photoacoustic spectra were measured for a P $r^{3+}$ solution with a very low molar absorptivity. The detection sensitivity was 4.3 $\times$10$^{-5}$ c $m^{1}$ and was 100 times better than that of a UV/VIS spectrophotometer. The Eu(III) excitation spectra($^{7}$ $F_{0}$ longrightarrow $^{5}$ $D_{0}$ transition) were measured for Eu(III)-phthalate complexes using laser fluorescence spectroscopy, showing that only two species, 1:1 and 1:2 complexes, are present in the Eu(III)-phthalic acid system. The size and size distribution for colloidal humic acids and Eu(III)-humate colloids was determined using photon correlation spectroscopy. The presence of Eu(III) enhanced the aggregation of humic acids.s.

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Europium-Enoxacin Complex as Fluorescence Probe for the Determination of Folic Acid in Pharmaceutical and Biological Samples

  • Alam, Al-Mahmnur;Kamruzzaman, Mohammad;Lee, Sang-Hak;Kim, Young-Ho;Min, Kyung
    • Bulletin of the Korean Chemical Society
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    • v.33 no.9
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    • pp.3055-3060
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    • 2012
  • A simple, rapid and sensitive spectrofluorometric method was developed for the determination of folic acid (FA), based on its quenching effect on the fluorescence intensity of enoxacin (ENX)-europium ($Eu^{3+}$) complex as a fluorescent probe. Fluorometric interaction between ENX-$Eu^{3+}$ complex and FA was studied using UV-visible and fluorescence spectroscopy. The quenched fluorescence intensity at an emission wavelength of 614 nm was proportional to the concentration of FA. Optimum conditions for the determination of FA were investigated. Under optimal conditions, the reduced fluorescence intensity at 614 nm was responded linearly with the concentration of FA. The linearity was maintained in the range of $1.25{\times}10^{-9}$ to $1.50{\times}10^{-7}$ M (R = 0.9986) with the limit of detection ($3S_b/m$) (where $S_b$ is the standard deviation of blank and m is the slop of linear calibration curve) of $6.94{\times}10^{-10}$ M. The relative standard deviation (RSD) for 9 repeated measurements of $1.0{\times}10^{-9}$ M FA was 1.42%. This method was simple, cost effective, and relatively free of interference from coexisting substances. Successful determinations of FA in pharmaceutical formulation and biological samples with the developed method were demonstrated.

Non-Invasive Environmental Detection using Heat Shock Gene-Green Fluorescent Protein Fusions

  • Cha, Hyeong-Jun
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.355-356
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    • 2000
  • Three 'stress probe' plasmids were constructed and characterized which utilize a green fluorescent protein (CFP) as a non-invasive reporter to elucidate Escherichia coli cellular stress responses in quiescent or 'resting' cells. Facile detection of cellular stress levels was achieved by fusion of three heat shock stress protein promoter elements, those of the heat shock transcription factor ${\sigma}^{32}$, pretense subunit ClpB, and chaperone DnaK, to the reporter gene $gfp_{uv}$. When perturbed by chemical or physical stress (such as heat shock, nutrient (amino acid) limitation, addition of IPTG, acetic acid, ethanol, phenol, antifoam, and salt (osmotic shock), the E. coli cells produced GFPuv which was easily detected from within the cells as emitted green fluorescence. A temporal and amplitudinal mapping of these responses was performed, demonstrating regions where quantitative delineation of cell stress was afforded.

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