• Title/Summary/Keyword: Chemiluminescence method

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A Membrane-Array Method to Detect Specific Human Intestinal Bacteria in Fecal Samples Using Reverse Transcriptase-PCR and Chemiluminescence

  • KIM PYOUNG IL;ERICKSON BRUCE D;CERNIGLIA CARL E.
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.310-320
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    • 2005
  • A membrane-based oligonucleotide array was used to detect predominant bacterial species in human fecal samples. Digoxygenin-labeled 16S rDNA probes were generated by PCR from DNA that had been extracted from fecal samples or slurries. These probes were hybridized to an array of 120 oligonucleotides with sequences specific for 40 different bacterial species commonly found in human feces, followed by color development using an alkaline phosphatase-conjugated antibody and NBT /BCIP. Twenty of the species were detected by this method, but E. coli, which was present at $\~$1 $\times 10$^5$ CFU per gram feces, was not detected. To improve the sensitivity of this assay, reverse transcriptase-PCR was used to generate probes from RNA extracted from fecal cultures. Coupled with a chemiluminescence detection method, this approach lowered the detection limit for E. coli from $\~1$ $\times 10$^6$ to ${\leq}$ 1 $\times 10$^5$ These results indicate that the membrane-array method with reverse transcriptase-PCR and chemiluminescence detection can simultaneously identify bacterial species present in fecal samples at cell concentrations as low as${\leq}$ 1 $\times 10$^5$ CFU per gram.

A Metal Enhanced Flow-Injection Chemiluminescence Method for the Rapid Determination of Norfloxacin in Pharmaceutical Formulations and Serum Sample

  • Kamruzzaman, Mohammad;Ferdous, Taslima;Alam, Al-Mahmnur;Lee, Sang-Hak;Kim, So-Yeun;Kim, Young-Ho;Kim, Sung-Hong
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.639-644
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    • 2011
  • A simple and highly sensitive chemiluminescence method to determine norfloxacin (NFLX) has been proposed by measuring the chemiluminescence (CL) intensities using a flow injection (FI) system. The CL intensity of the luminol-$H_2O_2$ system is strongly enhanced by the addition of Cu (II) in alkaline condition. The CL intensity is substantially increased after the injection of NFLX into the luminol-$H_2O_2$-Cu (II) system. The enhancement effect is attributed to a catalytic effect of Cu (II) due to the interaction with NFLX which forms a complex with the catalyst. Under the optimal conditions, the sensitizing effect of the CL intensity is proportional to the concentration of NFLX in the range of $1.5{\times}10^{-9}-5.9{\times}10^{-7}molL^{-1}$ (r = 0.9994) with a detection limit ($3{\sigma}$) of $2.98{\times}10^{-10}molL^{-1}$. The proposed method had good reproducibility with the relative standard deviation (RSD, n = 5) of 1.6% for $1{\times}10^{-7}molL^{-1}$ of NFLX. The possible reaction mechanism of the CL reaction is also discussed. This method has been successfully applied for the determination of trace amount of NFLX in pharmaceutical preparations and serum samples.

A Chemiluminescence Method for the Determination of Ascorbic Acid based upon Its Reaction with Cerium(IV) in the Presence of $Ru(bipy)_3^{2+}$

  • Lee, Sang-Hak;Bae, Hyun-Sook;karim, Mohammad-Mainul;Kim, Chang-Jin;Kim, Young-Sun;Lee, Hyun-Sook;Hong, Sang-Beom
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.149-153
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    • 2005
  • A chemiluminescence (CL) method for the determination of ascorbic acid was proposed, which is based on the enhancement of the CL intensity of $Ru(bipy)_3^{2+}$ - Ce (IV) system with the addition of ascorbic acid. The CL intensity is proportional to the concentration of ascorbic acid in the range of $1{\times}10^{-10}\;to\;1{\times}10^{-5}M$ with a correlation coefficient of 0.9985. The detection limit is $3{\times}10^{-11}M$. The relative standard deviation (R.S.D.) of 10 determinations of 0.1 mM ascorbic acid was 1.75%. The proposed method has been successfully applied to the determination of ascorbic acid in the commercia] drinks. The possible mechanism of the CL reaction was studied.

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Chemiluminescence Determination of Balofloxacin Based on Europium (III)-Sensitized KBrO3-Na2S2O4 Reaction in Micellar Medium

  • Zhao, Fang;Qi, Yu;Xiong, Wei
    • Bulletin of the Korean Chemical Society
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    • v.33 no.1
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    • pp.204-208
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    • 2012
  • A novel chemiluminescence (CL) flow injection method for the determination of balofloxacin is described. The method is based on the weak CL signal arising from the reaction of $KBrO_3$ with $Na_2S_2O_4$ in acidic medium being significantly enhanced by balofloxacin in the presence of europium (III) ion and sodium dodecyl benzene sulfonate (SDBS). The experimental conditions that affected CL intensity were carefully optimized and the CL reaction mechanism was briefly discussed. Under the optimum conditions, the relative CL intensity was proportional to the concentration of balofloxacin in the range of $7.0{\times}10^{-11}$ to $3.0{\times}10^{-7}g\;mL^{-1}$. The detection limit was $2.7{\times}10^{-11}g\;mL^{-1}$ and the relative standard deviation was 2.1% for $7.0{\times}10^{-10}g\;mL^{-1}$ balofloxacin (n = 13). The proposed method was successfully applied to the determination of balofloxacin in pharmaceutical formulations and biological fluids.

Experimental Study on Heat Release in a Lean Premixed Dump Combustor using OH Chemiluminescence Images (희박 예혼합 덤프 연소기에서 OH 자발광을 이용한 열 방출에 관한 실험적 연구)

  • Moon, Gun-Feel;Lee, Jong-Ho;Jeon, Chung-Hwan;Chang, Young-June
    • Proceedings of the KSME Conference
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    • 2004.11a
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    • pp.1146-1151
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    • 2004
  • Measurements of OH chemiluminescence in an atmospheric pressure, laboratory-scale dump combustor at equivalence ratios ranging from 0.63 to 0.89 were reported. The signal from the first electronically excited state of OH to ground state was detected through a band-pass filter with an ICCD. The objectives of this study are two: One is to see the effects of equivalence ratio on global heat release rate and local Rayleigh index distribution. To get the local Rayleigh index distribution, the line-of-sight images were inverted by tomographic method, such as Abel de-convolution. Another aim is to investigate the validity of using OH chemiluminescence acquired with an ICCD as a qualitative measure of local heat release. For constant inlet velocity and temperature, the overall intensities of OH emission acquired at different equivalence ratio showed periodic and higher value at high equivalence ratio. OH intensity averaged over one period of pressure increased exponentially with equivalence ratio. Local Rayleigh index distribution clearly showed the region of amplifying or damping the combustion instability as equivalence ratio increased. It could provide an information/insights on active control such as secondary fuel injection. Finally, local heat release rate derived from reconstructed OH images were presented for typical locations.

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Determination of Horseradish Peroxidase (HRP) using an Enhanced Chemiluminescence Assay (증강 화학발광 기법을 이용한 horseradish peroxidase(HRP)의 검량)

  • Kim, Wongee;Kim, Keunhan;Lee, Seungmok
    • Journal of Korean Society on Water Environment
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    • v.25 no.1
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    • pp.84-89
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    • 2009
  • Our nation's water resources remain susceptible to contamination by phenolic agrichemicals. These compounds can be toxic to a variety of organisms including humans. Their disposal is restricted in many countries with strict limits for acceptable concentrations in drinking water. Enzyme-mediated in situ stabilization has been advocated as an approach for the treatment of phenolic compounds in soils and groundwater. This study reports the development of a new approach to quantify the activity of the HRP enzyme in aqueous systems. The method is based on the coupled processes of energy transfer and enhanced chemiluminescence using a luminol-$H_2O_2$-HRP system. In this study, the effects of solution pH, ionic strength and aqueous concentrations of HRP, $H_2O_2$ and enhancer were evaluated on the p-iodophenol-enhanced, HRP-catalyzed chemiluminescence reaction intensity in Tris-HCl buffer. All assay components were found to affect the maximum chemiluminescene intensity. The calibration curve for HRP showed the linear relationship with maximum light intensity.

Determination of $Cu^{2+}$ by Lophine Chemiluminescence

  • Kim Young-Sun;Karim Mohammad Mainul;Lee Sang-Hak;Choi Kyoung-Hye;Choi Jong-Ha;Lee Sung-Ho
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.137-141
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    • 2005
  • The chemiluminescence reaction of lophine with $H_2O_2$ in alkaline solution has been investigated for use in determination of $Cu^{2+}$ ions. The observed chemiluminescence intensity is found to be a function of the concentration of $Cu^{2+}$. Under the optimum reagent concentrations such as $4{\times}10^{-4}M$ lophine, 0.8 M KOH, 0.2M $H_2O_2,{\lambda}_{em}$, 533nm, the linear range and the detection limit were found to be 0.048ug/ml-48.32ug/ml (R=0.99897) and 0.005ulg/ml respectively. Relative standard deviation for five determinations of 24.16ug/ml $Cu^{2+}$ is 2.35%. The interference from other species was investigated. The proposed method was applied to the determination of $Cu^{2+}$ in different water samples.

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Determination of Uric Acid by Chemiluminescence Measurement Using Tris(2,2'-bipyridine)ruthenium(II)-Octylphenylpolyglycol Ether System

  • Kim, Young-Sang;Park, Jeung-Hee;Choi, Yoon-Seok
    • Bulletin of the Korean Chemical Society
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    • v.25 no.8
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    • pp.1177-1181
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    • 2004
  • The determination of uric acid in urine samples was studied by a chemiluminescence measurement using tris (2,2'-bipyridine)ruthenium(II)-octylphenylpolyglycolether [Ru$(bpy)_3^{2+}$ -OPE] system. The oxidized uric acid by Ce(IV) excited Ru$(bpy)_3^{2+}$ to emit a chemiluminescence in this system so that the intensity was stoichio-metrically dependent upon the concentration of uric acid. In a reaction cell, a luminescent reagent, oxidant, surfactant and sulfuric acid were flowed into and mixed with a taken sample. Experimental conditions were optimized to obtain the maximum intensity of chemiluminescence. Each reactant solution of more than the following concentration gave a good result: $2\;{\times}\;10^{?4}$ M Ru$(bpy)_3^{2+}$ , 0.01 M Ce(IV), 6% OPE, and 0.33 M $H_2SO_4$. Any interferences were not shown in this process by the investigation of concomitant constitutes such as albumin, creatine, lactic acid, glucose, urea, $Cl^?,\; Mg^{2+},\;Ca^{2+}$ and so on. The linearity of a calibration curve was good with r = 0.998, the relative standard deviation of the slope was 3.3% and the detection limit was 5.6ng/mL. The recoveries of 80 to 91% were obtained from the standard spiked samples. The values were little bit low, but this procedure could be considered to be reliable for the determination of trace uric acid in urine samples.

Detection of Superoxide Anion and Singlet Oxygen in the Decomposition of Several Peroxovanadium(V) Complexes

  • Kanamori, Kan;Hata, Kaori;Shimoyama, Toshiyuki;Hayakawa, Shingo;Tajima, Hirotaka;Matsugo, Seiichi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.412-414
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    • 2002
  • Several peroxovanadium(V) complexes with an organic chelate ligand decompose spontaneously, depending on the nature of the chelate ligand. The self-decomposition reactions of the dinuclear peroxovanadium(V) complex with 2-oxo-l,3-diaminopropane-N,N,N',N'-tetraacetate (dpot) and the peroxovanadium(V) complexes with N-carboxymethylhistidinate (cmhist) and histamine-N,N-diacetate (histada) accompany the reduction of vanadium(V) to vanadium(IV). This implies that the peroxide anion acts as a reducing agent and thus the peroxide is oxidized in the decomposition process of the peroxovanadium(V) complexes. The oxidized dioxygen species have been characterized spectrophotometrically. Superoxide anion has been detected in 2-3 % yields using the reduction of cytochrome c method and chemiluminescence method utilized MCLA as a fluorescer. Singlet oxygen has also been detected in higher yields on the basis of chemiluminescence of tryptophan.

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A Sensitive Procedure for the Rapid Determination of Mandelic Acid by Flow Injection Analysis and Chemiluminescence Detection

  • Alam, Seikh Mafiz;Jeon, Chi-Wan;Karim, Mohammad Mainul;Lee, Sang-Hak;Wabaidur, Saikh Mohammad;Suh, Yeoun-Suk;Chung, Hye-Young
    • Bulletin of the Korean Chemical Society
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    • v.30 no.1
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    • pp.102-106
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    • 2009
  • A rapid and sensitive chemiluminescence (CL) method using flow-injection (FI) has been developed for the determination of mandelic acid which is based on the enhancement of mandelic acid to the CL intensity of ${Ru(bipy)_{3}}^{2+}$-Ce(IV) system. The enhancement effect was dependent on the concentration of mandelic acid, based on which, CL system was established for the determination of mandelic acid. The concentrations of ${Ru(bipy)_{3}}^{2+}$+, Ce(IV), and $H_2SO_4$ were optimized. Under the optimum experimental conditions, the linear range and detection limit are 1.46-342.0 ${\mu}g/ml$ and 0.072 ${\mu}g/ml$, respectively. The correlation coefficient (R) was 0.99732. The utility of this method was demonstrated by determining mandelic acid in capsules and human urine sample.