• Title/Summary/Keyword: luminol chemiluminescence assay

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Chemiluminescence immunochromatographic analysis for the quantitative determination of algal toxins

  • Pyo, Dongjin;Kim, Taehoon
    • ALGAE
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    • v.28 no.3
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    • pp.289-296
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    • 2013
  • For the quantitative detection of algal toxin, microcystin, a chemiluminescence immunochromatographic assay method was developed. The developed system consists of four parts, chemiluminescence assay strip (nitrocellulose membrane), horse radish peroxidase labeled microcystin monoclonal antibodies, chemiluminescence substrate (luminol and hydrogen peroxide), and luminometer. The performance of the chemiluminescence immunochromatographic assay system was compared with high performance liquid chromatography (HPLC) detection. The detection limit of chemiluminescence immunochromatographic assay system is several orders of magnitude lower than with HPLC. The chemiluminescence immunochromatography and HPLC results correlated very well with the correlation coefficient ($r^2$) of 0.979.

Development of Novel Method for the Detection of Microcystin Using Chemiluminescence Immunochromatography

  • Pyo, Dong-Jin;Yoo, Ji-Sun
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.149-152
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    • 2011
  • A new chemiluminescence immunochromatographic analysis system with high sensitivity and high reproducibility was developed for the determination of microcystins (MCs) in water. Horse radish peroxidase (HRP) labeled microcystin monoclonal antibody was used for the sensitive chemiluminescence detection. The chemiluminescence immunochromatographic analysis system was composed of microcystin LR (MCLR)-monoclonal antibody (mAb)-Horse Radish Peroxidase (HRP) conjugate, MCLR-BSA conjugate, luminol, hydrogen peroxide mixture solution, an immunochromatographic assay strip and luminometer. To detect the concentration of microcystins in water, we utilized one spot analysis of the strip instead of flow type analysis. We could detect the microcystins in water at a concentration as low as 9.45 pg/mL with the chemiluminescence (CL) detection.

Comparison of Spectrophotometric, HPLC and Chemilumines­cence Methods for 3-Nitrotyrosine and Peroxynitrite Interaction

  • Turan Nilufer Nermin;Ark Mustafa;Demiryurek Abdullah Tuncay
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.358-363
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    • 2005
  • We have studied the interaction of 3-nitrotyrosine with peroxynitrite using three different methods; chemiluminescence, spectrophotometry and HPLC. Peroxynitrite-induced luminol or lucigenin chemiluminescence were significantly decreased by 3-nitrotyrosine, in concentration­dependent manners. The intensity of the peroxynitrite spectrum was also markedly reduced in the presence of 3-nitrotyrosine in the spectrophometric assay. However, there was no attenuation of the 3-nitrotyrosine signal in the HPLC assay after mixing with peroxynitrite. The interaction of 3-nitrotyrosine and hypochlorous acid (HOCI) was also studied via the chemilumines-cence assay, where the HOCI-induced responses were markedly inhibited by 3-nitrotyrosine. These results suggest that caution should be taken when studying the levels or interactions of 3-nitrotyrosine.

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 aromatic amino acids by chemiluminometric assay with Luminol-H2O2-Cu(II) system (Luminol-H2O2-Cu(II) 시스템을 이용한 방향족 아미노산의 화학발광법적 정량)

  • Kim, Kyung-Min;Kim, Young-Ho;Lee, Sang-Hak
    • Analytical Science and Technology
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    • v.25 no.3
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    • pp.171-177
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    • 2012
  • A determination method of aromatic amino acids such as trytophan (Trp), tyrosine (Tyr), and phenylalanine (Phe) using luminol-$H_2O_2$-Cu(II) system has been presented. In the presence of an aromatic amino acid, the enhanced chemiluminescence (CL) intensity of luminol-$H_2O_2$-Cu(II) system was obtained by forming a complex between Cu(II) and the amino acid. Based on the above phenomenon, a sensitive and fast determination of three aromatic amino acids was performed using the CL method in batch-type detection system. To optimize determination conditions, the kinetic influence of an aromatic amino acid on the luminol-$H_2O_2$-Cu(II) system and the effects of $H_2O_2$ and Cu(II) concentration, pH, and buffers were investigated. Under the optimized conditions, the calibration curve was linear over the range from $1.0{\times}10^{-6}$ to $2.0{\times}10^{-5}\;M$ for Trp, $1.0{\times}10^{-6}$ to $2.0{\times}10^{-5}\;M$ for Try, and $2.0{\times}10^{-6}$ to $2.0{\times}10^{-5}\;M$ for Phe, respectively. In this range, reproducibility (RSD, n = 4) of Trp, Try, and Phe were 3.21%, 2.64%, and 2.48%, respectively. The limit of detection ($3{\sigma}/s$) was calculated to be $6.8{\times}10^{-7}\;M$ for Trp, $5.7{\times}10^{-7}\;M$ for Try, and $9.6{\times}10^{-7}\;M$ for Phe.

Comparison of anti-oxidant activities of seventy herbs that have been used in Korean traditional medicine

  • Ko, Seong-Hee;Choi, Seong-Won;Ye, Sang-Kyu;Yoo, Sang-Ho;Kim, Hyun-Sook;Chung, Myung-Hee
    • Nutrition Research and Practice
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    • v.2 no.3
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    • pp.143-151
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    • 2008
  • Many herbs have been used as therapeutics in Korean traditional medicine. In view of their clinical indications, anti-oxidant activity may contribute to their pharmacological effects. However, anti-oxidant information on these plants has not been available. In this study, seventy herbs which have been used in Korean traditional medicine were selected and screened for anti-oxidant activity using their water extracts. The anti-oxidant activity was assessed by their ability to inhibit three oxidation reactions; luminol/Fenton reagent, 2, 7-dichlorodihydrofluorescein(DCHF)/Fenton reagent and DCHF/peroxynitrite. In each assay, 70 herbs were divided into two groups; anti-oxidant group which inhibited the respective oxidation reaction and was majority(about 60 herbs), and pro-oxidant group which enhanced the oxidation reaction but was minority(more or less 10 herbs). When the herbs were listed in the order of their anti-oxidant strength, the orders obtained from each assay were found to be quite similar. The upper top rankers(more or less 10 herbs) in each assay showed strong activity compared to the others. The uppermost rankers in each assay were Rubus coreanus Miquel/Rubus schizostylus(覆盆子), Schisandra chinensis Baillon/Schizandra chinensis(五味子) and Terminalia chebula Retziusl Terminalia chebula(訶子). Of the pro-oxidant herbs, about 4-5 herbs were strongly pro-oxidant, which enhanced the control oxidation reactions to 150-300%. But the meaning of this observation is not known since few of them in one assay were also anti-oxidant in other assays. The results obtained in the present study may serve as information for understanding pharmacological effects of these herbs and developing new drugs from them.

Screening of Effective Extraction Conditions for Increasing Antioxidant Activities of Licorice Extracts from Various Countries of Origin (원산지별 감초추출물의 항산화활성 증가를 위한 효율적인 추출조건 탐색)

  • Ha, Ji Hoon;Lee, Hye Mi;Kwon, Soon Sik;Kim, Hae Soo;Kim, Moon Jin;Jeon, So Ha;Jeong, Yoo Min;Hwang, Jun Pil;Park, Jong-Ho;Choi, Yung-Key;Park, Jino;Park, Soo Nam;Park, Dong-Sik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.4
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    • pp.259-269
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    • 2013
  • In this work, licorice extracts were prepared using various extraction conditions such as extraction solvent, temperature, and time from Glycyrrhiza uralensis (G. uralensis) produced in Korea and China and Glycyrrhiza glabra (G. glabra) in Uzbekistan. The optimum extraction condition was selected from the extraction yields and antioxidative activities of extracts. Korea licorice extracts showed the highest free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity (46.05%) under the extraction condition of 85% ethanol at $60^{\circ}C$ for 6 hours. The prominent ROS (reactive oxygen species) scavenging activity using luminol-dependent chemiluminescence assay and the cellular protective effect against $^1O_2$ induced cellular membrane damage were also shown from the extracts obtained from the same condition. Especially, Korea G. uralensis extracts exhibited the higher prominent protective effect (${\tau}_{50}$ = 116.4 min) than (+)-(+)-${\alpha}$-tocopherol (${\tau}_{50}$ = 28.5 min) and the extraction yield of Korea licorice extract was 18.75%, which is 1.2 times and 2.5 times higher than that of Uzbekistan and China, respectively. These results indicate that the extraction condition of 85% ethanol at $60^{\circ}C$ for 6 hours is optimal to prepare licorice extracts, which can be applicable as anti-oxidative cosmetic materials.

A Hyperactive Neutrophil Phenotype in Aggressive Periodontitis

  • Kim, Kap-Yul;Kim, Min-Kyung;Choi, Yun-Sik;Kim, Yong-Cheol;Jo, Ah-Ram;Rhyu, In-Chul;Choi, Young-Nim
    • International Journal of Oral Biology
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    • v.37 no.2
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    • pp.69-75
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    • 2012
  • Although neutrophils function in both defense and tissue destruction, their defensive roles have rarely been studied in association with periodontitis. We hypothesized that peripheral neutrophils are pre-activated in vivo in periodontitis and that hyperactive neutrophils would show enhanced phagocytic ability as well as an increased production of reactive oxygen species (ROS). Peripheral blood neutrophils from patients with aggressive periodontitis and age/gender-matched healthy subjects (10 pairs) were isolated. The levels of CD11b and CD64 expression on the neutrophils and the level of plasma endotoxin were determined by flow cytometry and a limulus amebocyte lysate test, respectively. In addition, neutrophils were subjected to a flow cytometric phagocytosis assay and luminol-enhanced chemiluminescence for non-opsonized Fusobacterium nucleatum in parallel. The neutrophilsfrom most patients expressed increased levels of both CD11b and CD64. In addition, the plasma from these patients tended to contain a higher level of endotoxin than the healthy controls. In contrast, no differences were found between the two groups with regard to phagocytosis or ROS generation by F. nucleatum. The ability to phagocytose F. nucleatum was found to positively correlate with the ability to produce ROS. In conclusion, peripheral neutrophils from patients with aggressive periodontitis are hyperactive but not hyperreactive to F. nucleatum.

Antioxidative and Antiaging Effects of Persicaria hydropiper L. Extracts (여뀌 추출물의 항산화 및 항노화에 관한 연구)

  • Kim, Eun-Hee;Kim, Jung-Eun;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.4
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    • pp.293-300
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    • 2009
  • In this study, we investigated the antioxidative activity and inhibitory effects on elastase and tyrosinase of Persicaria hydropiper L. extracts. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of ethyl acetate fractions of Persicaria hydropiper L. was $5.23\;{\mu}g/mL$. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Persicaria hydropiper L. extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol - dependent chemiluminescence assay. The ROS scavenging activities ($OSC_{50}$) of ethyl acetate fractions of Persicaria hydropiper L. was $0.40\;{\mu}g/mL$. The protective effects of extract / fractions of Persicaria hydropiper L. on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Persicaria hydropiper L. extracts suppressed photohemolysis in a concentration dependent manner ($1\;{\sim}\;10\;{\mu}g/mL$). Inhibitory effects ($IC_{50}$) on tyrosinase of aglycone fraction of Persicaria hydropiper L. extracts was $8.90\;{\mu}g/mL$. Inhibitory effects ($IC_{50}$) on elastase of aglycone fraction of Persicaria hydropiper L. extracts was $2.37\;{\mu}g/mL$. These results indicate that extract / fractions of Persicaria hydropiper L. can function as antioxidants in biological systems, particularly skin exposed to UV radiation by anti-oxidative activity and protect cellular membranes against ROS. Persicaria hydropiper L. extract / fractions could be used as a new cosmeceutical for whitening and anti-wrinkle products.

Evaluation of Antioxidant Potential of Extract/Fractions of Equisetum arvense (I) (쇠뜨기 추출물의 항산화 작용에 관한 연구(I))

  • Park, Soo-Nam;Yang, Hee-Jung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.2
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    • pp.61-67
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    • 2007
  • In this study, the antioxidative effects of Equisetum arvense extracts were investigated. The free radical (1,1 diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract/fractions of Equisetum arvense was in the order: 50 % ethanol extract ($182.04{\mu}g/mL$) < ethylacetate fraction ($54.50{\mu}g/mL$) < deglycosylated flavonoid aglycone fraction ($14.13{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Equisetum arvense extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol- dependent chemiluminescence assay. The order of ROS scavenging activity was deglycosylated flavonoid aglycone fraction ($OSC_{50}$, $3.54{\mu}g/mL$) < 50 % ethanol extract ($0.80{\mu}g/mL$) < ethylacetate fraction ($0.006{\mu}g/mL$). Ethylacetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Equisetum arvense on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethanol extract (50%) suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated aglycone extract exhibited the most prominent celluar protective effect ($\tau_{50}$, 161.10 min at $10{\mu}g/mL$). These results indicate that extract/fractions of Equisetum arvense can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS.