• Title, Summary, Keyword: Spin trapping ESR

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Enhancing Effect of Organic Substances on Hydroxyl Radical Generation During Ozonation of Water: Stopped-Flow ESR Technique

  • Han, Sang-Kuk
    • Bulletin of the Korean Chemical Society
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    • v.25 no.12
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    • pp.1907-1910
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    • 2004
  • Generation of hydroxyl radical, one of their major active species in ozonation of water was directly observed with spin-trapping/electron spin resonance (ESR) technique using 5,5-dimethyl-pyrrolidine-1-oxyl (DMPO) as a spin-trapping reagent. Hydroxyl radical was trapped with DMPO as a stable radical, DMPO-OH. 80 mM of ozone produced $1.08{\times}10^{-6}$M of DMPO-OH, indicating that 1.4% of ${\cdot}$OH is trapped with DMPO if ${\cdot}$OH is produced stoichiometrically from ozone. Humic acid suppressed DMPO-OH generation in a dose-dependent manner. Generation rate of DMPO-OH was determined with ESR/stopped-flow measurement. Phenol derivatives increased the amount and generation rate of DMPO-OH, indicating that phenol derivatives enhance·OH generation during ozonation of water.

Determination of Free Radicals in Mainstream Cigarette Smoke by Electron Spin Resonance (전자스핀공명에 의한 담배연기 중 자유라디칼 측정)

  • Lee, Jeong-Min;Lee, John-Tae;Park, Jin-Won;Hwang, Keun-Joong
    • Journal of the Korean Society of Tobacco Science
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    • v.29 no.2
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    • pp.146-151
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    • 2007
  • Gas phase and particulate phase radicals in mainstream cigarette smoke were determined Electron Spin Resonance(ESR) spectroscopy. The free radicals in particulate phase have been investigated by benzene extract of Cambridge Filter Pad containing the smoke condensate. Spin trapping method in conjunction with ESR was used to investigate free radicals in the gas phase of cigarette smoke. Several analytical experiments were conducted in order to determine the optimal conditions for maximum signal intensities and reproducibility of results. All the tests were optimized and normalized using the University of Kentucky 2R4F reference cigarette. The optimal conditions were 0.6 mL for analysis volume of ESR, $4{\sim}5\;mL$ for collection volume of spin-adducts, and PBN for quantification of free radicals in gas phase. The radical levels of Kentucky 2R4F cigarettes were found $2.18{\times}10^{14}\;spins/cig.$ and $2.10{\times}10^{15}\;spins/cig.$ in gas phase.

ESR-Spin Trapping Detection of Radical Center Formed on the Reaction of Metmyoglobin with Hydrogen Peroxide

  • Jeong, Sang-Hyeon;Hong, Sun-Joo
    • BMB Reports
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    • v.28 no.4
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    • pp.293-300
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    • 1995
  • The radical centers detected in the reaction of metmyoglobin (MetMb) with hydrogen peroxide ($H_2O_2$) have been studied by using a spin trapping technique. A broad 5-line asymmetric electron spin resonance (ESR) spectrum, with $2A_{max}=4.07\;mT$ and $2A_{min}=2.97\;mT$, obtained after incubation of MetMb with $H_2O_2$ in the presence of a spin trap, 5,5-dimethyl pyrroline-N-oxide (DMPO) was gradually weakened with time and disappeared completely by 6 min after addition of guanidine-HCl (14 M). When a higher concentration (6 M) of the agent was added, the signal disappeared within 40 see and the DMPO/OH signal appeared immediately. Then, a new 8-line signal with similar intensities grew gradually and was fixed by 45 min, coexisting with the DMPO/OH signal. This new signal was found to be composite, consisting of two different radical species. One of the 6-line signals, with $a_N$ 1.49 mT and $a_H$ 0.988 mT, was assigned to the DMPO/phenoxyl radical adduct. The second 6-line signal with $a_N$ 1.55 mT and $a_H$ 2.22 mT was assigned to carbon-centered radical adduct. When 3,3,5,5-tetramethylpyrrolin-N-oxide (TMPO), was employed in the place of DMPO, another broad asymmetric 5-line signal was detected with $2A_{max}=3.99\;mT$ and $2A_{min}=3.04\;mT$, which is virtually identical to that obtained from the DMPO system The shape of the spectrum of the TMPO adduct changed drastically, with lapse of time resulting in a broad singlet after 40 min. The broad singlet was assigned to the porphyrin radical adduct. Incubation of globin with Fenton reagent in the presence of DMPO initially gave a DMPO/OH signal. Then, a new 12-line signal began to grow after one minute and fixed after 15 min. coexisting with the DMPO/OH signal, This 12-line signal was assigned to DMPO/phenoxyl with $a_N$ 1.47 mT, $a_{{\beta}H}$ 0.99 mT and $a_{{\gamma}H}$ 0.13 mT. A minor concentration of carbon-centered radical adduct was also detected. This radical composition is identical to that of guanidine HCl treated MetMb/DMPO/$H_2O_2$ system, indicating that the radical producing conditions are somehow common in both systems. Heme iron can be released by excess $H_2O_2$ in the MetMb/$H_2O_2$ system, providing for Fenton reagent. When MetMb was pretreated with tyrosine blocking agent, $KI_3$ the broad 5.line MetMb-derived signal was not detected in the MetMb/DMPO/$H_2O_2$ system, whereas no such effect was detected on such system of Hb in which the radical center was assigned to cysteine residue not tyrosine, indicating that tyrosine residue is a main radical center produced in the MetMb/$H_2O_2$ system Thus, the present data strongly support the previous indication that the apomyoglobin-derived radical center formed in the reaction of MetMb with $H_2O_2$ is a tyrosine residue.

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Antioxidant effect on Aspalatone on the toxicity induced by Doxorubicin and by Fenton type condition (Doxorubicin과 Fenton 형의 반응에 의해 유도된 독성에 미치는 Aspalatone의 항산화 작용)

  • Kim, Jin;Nam, Sung-Won;Seo, Jung-Hye;Han, Sang-Hwa;Gu, Chang-Hwi;Kim, Hyung-Chun
    • Proceedings of the Korean Society of Applied Pharmacology
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    • pp.94-94
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    • 1995
  • Aspalatone (APT)의 예상되는 항산화 작용을 검색하기 위하여 Doxorubicin(Dox.) 유발독성 (Dox.의 작용 발현 기전은 oxyradical 생성에 의해 매개됨)으로 인하여 변화되는 항산화 효소계 및 과산화지질 생성에 대하여 APT의 효과를 검색하고, 별도로 Hydroxyl radical 생성에 미치는 APT의 효능을 검정하기 위하여 ESR spin trapping technique을 이용하여 ㆍOH/DMPO ESR signal을 검정하여 아래의 결과를 얻었다. Dox 투여 2일 후에 생쥐 심장에서 얻어진 SOD-1의 보상성 유도는 APT 병용 투여로 억제되었고, Catalase 활성을 유도하였으나 MBA 치에는 유의한 변화가 나타나지 않았다. 반면에 간에서는 APT으로 인하여 GSH-PX가 현저히 유도되었고 (다른 항산화 효소에는 유의한 변화가 나타나지 않았음), MDA 치가 억제되었다. 한편 Fenton 반응동안 증가되었던 ㆍOH/DMPO ESR signal은 APT에 의해 억제되었다. 이상의 결과에서 APT의 항산화성 효능이 인정되지만, Dox. 및 APT 각각의 용량 및 노출 시간의 상관성, 그리고 생화학적인 결과에 대한 (미세)조직학적인 확인이 요구되며 각각의 실험군, 즉 aspirin, maltol에 대한 효능/효력 검정도 추가로 보완되어야 할 것으로 생각된다.

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The Study of Free Radical Scavenging Effect of Lycii Fructus by Liver Injury of Rats (백서 간손상에 의한 구기자의 유리자유기 소거능에 관한 연구)

  • Yoon Sang Ju;Jung Sun Yeong;Kim Young Mi;Ha Ki Tae;Kim Cheorl Ho;Kim Dong Wook;Kim June Ki;Choi Dall Yeong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.1
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    • pp.91-100
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    • 2003
  • In the present study, we investigated the protective effect of the Lycii Fructus water extracts (LFE) against CCl4-induced hepatotoxicity and the mechanism underlying these protective effects in the rats. The pretreatment of LFE has shown to possess a significant protective effect by lowering the serum alanine and aspartate aminoteansferase (AST and ALT) and alkaline phosphatase (ALP). This hepatoprotective action was confirmed by histological observation, In addition, the pretreatment of LFE prevented the elevation of hepatic malondialdehyde (MDA) formation and the depletion of reduced glutathione (GSH) content and catalase activity in the liver of CC1₄-injected rats. The LFE also displayed hydroxide radical scavenging activity in a dose-dependent manner (IC50 = 83.6 μg/ml), as assayed by electron spin resonance (ESR) spin-trapping technique. Moreover, the expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR), was significantly decreased in the liver of LFE-pretreated rats when compared with that in the liver of control group. Based on these results, it was suggested that the hepatoprotective effects of the LFE may be related to antioxidant effects and regulation of CYP2E1 gene expression.

The Study of Protective Effect of Puerariae Radix against $CC1_4$-induced Hepatotoxicity ($CC1_4$로 유발된 백서의 간손상에 대한 갈근의 간보호작용 연구)

  • Hyun Dong Hwan;Jung Sun Yeong;Jung Sang Shin;Ha Ki Tae;Kim Cheorl Ho;Kim Dong Wook;Kim June Ki;Choi Dall Yeong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.297-307
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    • 2003
  • In the present study, we investigated the protective effect of the Puerarie Radix water extract (PRE) against CCl₄-induced hepatotoxicity and the mechanism underlying these protective effects in the rats. The pretreatment of PRE has shown to possess a significant protective effect by lowering the serum alanine and aspartate aminoteansferase (AST and ALT) and alkaline phosphatase (ALP). This hepatoprotective action was confirmed by histological observation. In addition, the pretreatment of PRE prevented the elevation of hepatic malondialdehyde (MDA) formation and the depletion of reduced glutathione (GSH) content and catalase activity in the liver of CC1₄-injected rats. The PRE also displayed hydroxide radical scavenging activity in a dose-dependent manner (IC50 = 83.6 μg/ml), as assayed by electron spin resonance (ESR) spin-trapping technique. Moreover, the expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR), was significantly decreased in the liver of PRE-pretreated rats when compared with that in the liver of control group. Based on these results, it was suggested that the hepatoprotective effects of the PRE may be related to antioxidant effects and regulation of CYP2E1 gene expression.

Scavenging Effects of Tea Catechins on Superoxide and Hydroxy Radical

  • Park, Jaeil;Chen, Liuji;Yang, Xianqiang;Shen, Shengrong;Wang, Yuefei;Ho, Ryu-Beung
    • Journal of Life Science
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    • v.12 no.2
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    • pp.75-79
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    • 2002
  • Tea catechins, the most important compounds in tea polyphenols, can efficiently scavenge superoxide anion free-radical ($O_2$.), hydroxyl radical. (.OH) The mechanism of scavenging active oxygen free radicals was investigated by ESR spin trapping technique and Chemiluminescence. Results showed that various tea catechins constitute an antioxidant cycle in accordance with the decreasing order of the first reductive potential, and produce the effect of cooperative strength each other. Esterificated catechins could scavenge active oxygen free radicals more effectively than the non-esterificated ones. When.OH and $O_2$.- were scavenged by (-)-epigallocatechin gallate [(-)- EGCG], the stoichiometric factors were 6, and the rate constants of scavenging reaction reached $7.71{\times}10^6$ and $3.52{\times}10^{11}$ L $mmol^{-1}s^{-1}$, respectively. In the mean time, tea catechins could scavenge superoxide anion fiee radical ($O_2$-.) and hydroxyl radical (.OH) in a dose dependent manner. But at higher concentration or pH value, tea catechins can induce the prooxidant.

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Alaternin and Emodin with Hydroxyl Radical inhibitory and/or Scavenging Activities and Hepatoprotective Activity on Tacrine-Induced Cytotoxicity in HepG2 Cells

  • Jung, Hyun-Ah;Chung, Hae-Young;Takaka, Yokezawa;Kim, Youn-Chul;Hyun, Sook-Kyung;Choi, Jae-Sue
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.947-953
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    • 2004
  • The antioxidative and hepatoprotective potentials of two anthraquinones, alaternin (2-hydroxy-emodin) and emodin, to scavenge and/or inhibit hydroxyl radicals generated by the Fenton reaction and to protect tacrine-induced cytotoxicity in human liver derived HepG2 cells were evaluated, respectively. The inhibitory activity on hydroxyl radical generated in a cell-free chemical system (FeSO$_4$/$H_2O$$_2$) was investigated by a fluorescence spectrophotometer using a highly fluorescent probe, 2$^1$,7$^1$-dichlorofluorescein. The hydroxyl radical scavenging activity was determined by electron spin resonance spectroscopy using 5,5-dimethy-1-pyrroline-N-oxide as hydroxyl radicals trapping agents. Tacrine-induced HepG2 cell toxicity was determined by a 3-[4,5-dimethylthiazole-2yl]-2,5-diphenyltertrazolium bromide assay. Although the scavenging activity of alaternin on hydroxyl radical was similar to that of emodin in dose-dependent pat-terns, the inhibitory activity exhibited by the former on hydroxyl radical generation was stron-ger than that of the latter, with $IC_{50}$/ values of 3.05$\pm$0.26 $\mu$M and 13.29$\pm$3.20 $\mu$M, respectively. In addition, the two anthraquinones, alaternin and emodin showed their hepatoprotective activ-ities on tacrine-induced cytotoxicity, and the EC$_{50}$ values were 4.02 11M and 2.37 $\mu$M, respec-tively. Silymarin, an antihepatotoxic agent used as a positive control exhibited the EC$_{50}$ value of 2.00 $\mu$M. These results demonstrated that both alaternin and emodin had the simultaneous antioxidant and hepatoprotective activities.ies.

Improved photoelectric performance via fabricated heterojunction g-C3N4/TiO2/HNTs loaded photocatalysts for photodegradation of ciprofloxacin

  • Wu, Dongyao;Li, Jinze;Guan, Jingru;Liu, Chongyang;Zhao, Xiaoxu;Zhu, Zhi;Ma, Changchang;Huo, Pengwei;Li, Chunxiang;Yan, Yongsheng
    • Journal of Industrial and Engineering Chemistry
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    • v.64
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    • pp.206-218
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    • 2018
  • An Intercalated heterostructural $g-C_3N_4/TiO_2/HNTs$ supported photocatalyst was successfully prepared via sol-gel and calcination methods. The introduction of HNTs and the $g-C_3N_4-TiO_2$ heterojunction effectively enhanced the charge transfer and separation efficiency of photogenerated electron-hole pairs, which endued the $g-C_3N_4/TiO_2/HNTs$ hybrid material with an outstanding photoelectric performance and good stability. And an obviously enhanced photocatalytic activity was exhibited by photodegrading ciprofloxacin compared with pure $TiO_2$. Furthermore, the main active species were detected through trapping experiment and ESR spin-trap technique with DMPO, which confirmed that the $^{\bullet}O_2{^-}$ and the $h^+$ were the main active species in the photocatalytic system.