• ALGAE
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• 제28권3호
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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.

• Bulletin of the Korean Chemical Society
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• 제32권1호
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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.

• Archives of Pharmacal Research
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• 제28권3호
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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.

• 한국물환경학회지
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• 제25권1호
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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.

• 분석과학
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• 제25권3호
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• pp.171-177
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• 2012

Luminol-$H_2O_2$-Cu(II) 시스템을 이용한 화학발광법을 사용하여 방향족 아미노산인 트립토판, 타이로신 및 페닐알라닌의 정량분석을 하였다. 세 종류의 방향족 아미노산(트립토판, 타이로신, 페닐알라닌)을 luminol-$H_2O_2$-Cu(II) 시스템에 첨가하였을 때 아미노산이 존재하지 않을 때보다 화학발광세기가 더욱 증가하는 현상을 관찰하였으며, 이러한 현상을 이용하여 각 방향족 아미노산을 정량분석 하였다. 방향족 아미노산의 최적분석 조건을 조사하기 위하여, Cu(II) 이온 촉매에 의한 루미놀과 과산화수소의 화학발광반응에 미치는 아미노산의 반응속도론적 영향을 조사하였고 과산화수소와 Cu(II) 이온의 농도 그리고 pH와 완충용액의 영향을 조사하였다. 루미놀 화학발광 시스템의 최적 분석조건 하에서 수용액 중의 방향족 아미노산 정량분석을 위해 얻은 검정곡선에서 직선성이 성립하는 농도 범위는 각각 트립토판은 $1.0{\times}10^{-6}{\sim}2.0{\times}10^{-5}\;M$, 타이로신은 $1.0{\times}10^{-6}{\sim}2.0{\times}10^{-5}\;M$ 그리고 페닐알라닌은 $2.0{\times}10^{-6}{\sim}2.0{\times}10^{-5}\;M$이었으며, 이 구간에서 각 아미노산에 대한 상대표준편차(n = 4)는 순차적으로 3.21%, 2.64% 그리고 2.48%이었다. 그리고 각 아미노산의 검출한계($3{\sigma}/s$)는 트립토판 $6.8{\times}10^{-7}\;M$, 타이로신 $5.7{\times}10^{-7}\;M$, 페닐알라닌 $9.6{\times}10^{-7}\;M$이었다.

• Nutrition Research and Practice
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• 제2권3호
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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.

• 대한화장품학회지
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• 제39권4호
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• pp.259-269
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• 2013

본 연구에서는 한국 및 중국산 감초(Glycyrrhiza uralensis), 그리고 우즈베키스탄산 감초(Glycyrrhiza glabra)를 대상으로 추출용매, 추출온도, 추출시간 등 추출조건별 추출물을 제조하고 이들 추출물들의 추출 수율과 항산화 활성을 비교하여 최적의 추출조건을 선정하였다. 항산화 활성 중 자유 라디칼(1,1-phenyl-2-picrylhydrazyl, DPPH) 소거활성은 85% 에탄올로 $60^{\circ}C$에서 6 h 동안 추출한 조건에서 한국 감초로부터 얻은 추출물이 가장 높은 활성(46.05%)을 나타내었다. 루미놀 발광법을 이용한 추출물들의 총 항산화능 측정 실험과 피부 광노화에 중요한 $^1O_2$으로 유도된 세포막 손상에 대한 추출물들의 세포 보호 효과를 측정한 실험 모두에서도 위의 조건에서 가장 높은 항산화 활성을 나타내었다. 특히, 한국 감초는 ${\tau}_{50}$이 116.4 min으로 비교 물질인 (+)-${\alpha}$-tocopherol (28.5 min)보다 약 4 배나 높은 세포 보호 효과를 나타내었으며, 추출 수율은 18.75%로 우즈베키스탄 및 중국 감초보다 각각 1.2 배 및 2.5 배의 추출 수율을 나타내었다. 따라서, 본 연구 결과는 항산화 소재로 화장품에 응용하기 위하여 감초로부터 추출물을 얻기 위한 최적의 조건은 85% 에탄올로 $60^{\circ}C$에서 6시간 동안 추출하는 것임을 보여주었다.

• International Journal of Oral Biology
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• 제37권2호
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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.

• 대한화장품학회지
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• 제35권4호
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• pp.293-300
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• 2009

본 연구에서는 여뀌 추출물의 항산화, 미백, 항주름 효과를 통한 항노화 활성을 조사하였다. 여뀌 추출물의 free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거활성($FSC_{50}$)은 분획물 중 ethyl acetate 분획에서 $5.23\;{\mu}g/mL$로 가장 큰 활성을 나타내었다. Luminol-의존성 화학발광법을 이용한 $Fe^{3+}$-EDTA/$H_2O_2$계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 여뀌 추출물의 총 항산화능은 분획물 중 ethyl acetate 분획($0.40\;{\mu}g/mL$)이 큰 활성을 나타내었다. 여뀌 추출물에 대하여 rose-bengal로 증감된 사람 적혈구의 광용혈에 대한 억제 효과를 측정하였다. 여뀌 추출물의 경우 $1\;{\sim}\;10\;{\mu}g/mL$의 농도에서 광용혈을 억제하였다. 미백 및 주름억제 효과측정으로는 각각 tyrosinase 및 elastase의 활성 저해 효과를 측정하였다. Tyrosinase의 활성 저해 효과($IC_{50}$)는 aglycone 분획($8.90\;{\mu}g/mL$) 에서, elastase의 활성 저해 효과($IC_{50}$)도 aglycone 분획($2.37\;{\mu}g/mL$)에서 가장 큰 저해활성을 나타났다. 여뀌 추출물중 ethyl acetate 분획과 aglycone 분획에 대한 실험 결과로부터 항산화, 항노화 화장품 원료로서의 가능성을 확인하였다.

• 대한화장품학회지
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• 제33권2호
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• pp.61-67
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• 2007

본 연구에서는 쇠뜨기 추출물의 항산화능을 조사하였다. 추출물의 free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거활성($FSC_{50}$)은 50% 에탄올 추출물($182.04{\mu}g/mL$) < ethylacetate 분획(54.50) < 당을 제거시킨 플라보노이드 aglycone 분획(14.13) 순으로 증가하였다. Luminol-의존성 화학발광법을 이용한 $Fe^{3+}-EDTA/H_2O_2$ 계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 총항산화능은 당을 제거시킨 플라보노이드 aglycone 분획($OSC_{50}$, $3.54{\mu}g/mL$) < 50 % 에탄올 추출물(0.80) < ethylacetate 분획(0.006) 순으로 ethylacetate 분획에서 가장 큰 활성을 나타내었다. 쇠뜨기 추출물에 대하여 rose-bengal로 증감된 사람 적혈구의 광용혈에 대한 보호효과를 조사하였다. 50% 에탄올 추출물의 경우 농도 의존적($1{\sim}100{\mu}g/mL$)으로 광용혈을 억제하였다. 특히 ethylacetate 분획에서 당을 제거시킨 aglycone 추출물은 $10{\mu}g/mL$ 농도에서 $\tau_{50}$이 161.10min으로 매우 큰 세포보호 활성을 나타내었다. 이상의 결과들은 쇠뜨기 추출물이 $^1O_2$ 혹은 다른 ROS를 소광시키거나 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 가리킨다.