Fe(II)-EDTA chelate-induced aromatic hydroxylation of terephthalate as a new method for the evaluation of hydroxyl radical-scavenging ability.

The investigation of Fe(II)-EDTA chelate-induced aromatic hydroxylation of terephthalate in pH 7.4 phosphate buffer solution and a new method for the evaluation of hydroxyl radical-scavenging ability are reported. The method is based on attack of the hydroxyl radical on the terephthalate to produce highly fluorescent 2-hydroxyterephthalate, which is detected fluorimetrically. The formation of hydroxyl radical is believed to be the result of the reduction of molecular oxygen by Fe(II)-EDTA to form superoxide radical, which in turn dismutates to hydrogen peroxide, and then Fe(II)-EDTA catalyzes the decomposition of hydrogen peroxide to produce hydroxyl radical. The mechanism of the generation of hydroxyl radical in the proposed system was confirmed. This study established a simple and inexpensive method for the evaluation of the scavenging ability of some compounds on hydroxyl radicals.     

Determination of the antioxidant capacity of different food natural products with a new developed flow injection spectrofluorimetry detecting hydroxyl radicals

This paper presents an automatic spectrofluorimetric method (flow injection analysis spectrofluorimetry) for detecting hydroxyl radicals. Based on H₂O₂ catalyzed by Co²⁺ yielding HO, the method utilized sodium terephthalate to trap hydroxyl radicals and obtained sodium 2-hydroxyterephthalate by aromatic hydroxylation, which resulted in an increase in the fluorescence intensity. The relative fluorescence intensity was proportional to the concentration of hydroxyl radicals. Hydroxyl radicals could be determined indirectly, based on the increase of fluorescence. It was a simple, rapid, precise, sensitive and automatic technique for the determination of HO. The relative standard deviation of 11 determinations was 0.57%. The method can be used to sieve antioxidant medicines and will have theoretical and practical guidance on the mechanism of hydroxyl radicals-damaging biology.     

Formation of Oxygen Radicals in Solutions of Different 7,8‐Dihydropterins: Quantitative Structure‐Activity Relationships

Under certain conditions, 7,8‐dihydroneopterin in aqueous solution promotes hydroxyl‐radical formation. Thus, we investigated the stimulation of hydroxyl‐radical formation by ten different 7,8‐dihydropterins (=2‐amino‐7,8‐dihydropteridin‐4(1H)‐one), i.e., 6‐(1′‐hydroxy) derivatives 1 and 2 , methyl derivatives 3 – 7 , and 6‐(1′‐oxo) derivatives 8 – 10 . All but the 6‐(1′‐oxo) derivatives produced hydroxyl radicals, as measured by the amount of salicylic acid hydroxylation products. This amount was dependent on the stability of the dihydropterin used. In the presence of chelated iron ions, hydroxylation was increased in every case; even 6‐(1′‐oxo) derivatives showed a low hydroxylation of salicylic acid. The degree of increase, however, strongly depended on the side chain of the dihydropterin. The 7,8‐dihydroneopterin ( 2 ) was investigated in more detail. Iron ions influenced both, the stability of 2 and hydroxyl‐radical formation. While iron ions determined the kinetics of the reaction, the amount of 2 was responsible for the amount of hydroxyl radicals formed. Our data establish that promotion of hydroxyl‐radical formation by 7,8‐dihydropterins depends on the oxidizability of the dihydropterins and on their iron‐chelating properties.     

羟自由基在电极电解过程中的形成规律

对钛基二氧化铅电极电解过程中产生的^·OH进行了定量研究.结果表明,钛基二氧化铅电极在适当的电解条件下可以产生大量的自由基,碱性和高频条件下产生的自由基比酸性和低频条件下产生的自由基多.苯酚降解实验结果说明了其极强的氧化性能,在废水的深度处理方面有着极其广阔的应用前景.     

Electrochemically assisted Fenton reaction: reaction of hydroxyl radicals with xenobiotics followed by on-line analysis with high-performance liquid chromatography/tandem mass spectrometry

Oxygen radicals are generated in vivo by various processes, often as toxic intermediates in different metabolic transformations, and have been shown to play an important role for a large number of diseases. In this article we introduce an electrochemical flow-through system that allows generation of hydroxyl radicals for reaction with xenobiotics and subsequent detection of the oxidation products on-line with high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). The system is based on the Fenton reaction and is predominantly aimed at the generation of hydroxyl radicals; however, by minor variations to the system, a broad range of other radicals can be produced. Optimization of the system was performed with the radical scavenger 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). Under the same physical conditions, one injection through the electrochemical cell gave a higher yield of the oxidation product N-hydroxy-5,5-dimethylpyrrolidin-2-one than what was attained after 60 min with a chemical Fenton system catalyzed by ascorbic acid. Since the iron is added as Fe(3+), the initial mixture is 'inactive' until it reaches the electrochemical cell. This makes it very suitable for on-line analysis of the generated compounds, since the whole reaction mixture, including substrate, can be kept in a vial in an autosampler. The system described provides a useful tool for investigation of new radical scavengers and antioxidants. Since the hydroxyl radical adds readily to unsaturated pi-systems, the technique is also suitable for on-line generation and characterization of potential drug metabolites resulting from hydroxylation of double bonds and aromatic systems.     

2,2,6,6-四甲基哌啶-1-氧自由基促进的钒基催化剂催化苯直接氧化制苯酚

以钒基化合物为催化剂,在TEMPO(2,2,6,6-四甲基哌啶-1-氧自由基)存在下,能形成快速催化分子氧氧化苯制苯酚的催化体系.在反应过程中,由类似芬顿试剂反应过程生成的羟基自由基亲核进攻苯环,形成羟基环己二烯自由基;该羟基氢可在TEMPO存在的催化体系中消除,同时苯环氢可立即转移至氧原子而生成苯酚.在以[(CH3)...     

一种表征羟基自由基的新型荧光探针

表征羟基自由基 (· OH)的方法主要有电子自旋共振法[1] 和芳环羟基化法[2 ,3] 两大类 .电子自旋共振法灵敏度不高 ,且仪器设备昂贵 ,不适于常规分析 .芳环羟基化法操作较简单 ,灵敏度高 ,但芳环羟基化的产物往往不止一种 ,使得定量测定变得复杂 .其它方法如高效液相色谱法[4 ] ,化学发光法[5] 等也有报道 .顺磁性氮氧化合物能有效地清除自由基 [6 ,7] ,同时也是一种芳烃单重激发态的有效猝灭剂 [8,9] .当顺磁性氮氧化合物与荧光分子共价结合 ,所形成的荧光分子 -氮氧自由基复合物 (即自旋标记荧光分子 )仍保留对自由基反应的活性 ,但由于…     

An HPLC assay of hydroxyl radicals by the hydroxylation reaction of terephthalic acid

An HPLC assay for hydroxyl radicals is described. The hydroxyl radical was trapped by terephthalic acid (non-fluorescent), and 2-hydroxyl terephthalic acid (fluorescent) was quantitated by HPLC-fluorescence detection. At a terephthalic acid concentration of 4.25 mmol/L, the hydroxyl radical formed in the Fenton reaction was successfully assayed in the concentration range of hydrogen peroxide of 2.5-50 micro mol/L, where the concentration of Fe(II) was 50 micro mol/L. The fluorescence of 2-hydroxy terephthalate was stable at 24 h, and its detection limit by this method was 5 nmol/L (100 fmol).     

SPECTROSCOPIC STUDIES OF CUTANEOUS PHOTOSENSITIZING AGENTS–XII. SPIN TRAPPING STUDY OF THE FREE RADICALS GENERATED DURING THE PHOTOLYSIS OF PHOTO ALLERGENS BITHIONOL and FENTICHLOR

Free radicals were trapped and observed by ESR when photoallergens bithionol and fentichlor were irradiated in the presence of spin traps N- t -butyl-α-phenylnitrone (PBN) and 5,5-dimethyl-pyrroline-N-oxide (DMPO). In the absence of air, both PBN and DMPO trapped a carbon-centered radical. The carbon-centered radical, which was capable of abstracting a hydrogen atom from cysteine, glutathione, ethanol and formate, was identified as an aryl radical derived from the homolytic cleavage of the carbon-chlorine bond. In the presence of air, both carbon-centered radicals and hydroxyl radicals were trapped by DMPO. Under similar conditions, the yield of the hydroxyl radicals was greater from bithionol than from fentichlor. The presence of the hydroxyl radical was confirmed by kinetic experiments employing hydroxyl radical scavengers (ethanol, formate). Superoxide and H₂O₂ were not involved. Experiments with oxygen-¹⁷O indicated that the hydroxyl radicals came exclusively from dissolved oxygen. The precursor of the hydroxyl radical is postulated to be a peroxy intermediate (ArOO*) derived from the reaction of an aryl radical (Ar*) with molecular oxygen. Both bithionol and fentichlor photoionized only when excited in the UVC (<270 nm) region. Free radicals have long been postulated in the photodechlorination of bithionol and fentichlor and the present study provides supporting evidence for such a mechanism. Aryl and hydroxyl radicals are reactive chemical species which may trigger a series of events that culminate in photoallergy.     

Bisphenol A reactions with hydroxyl radicals: diverse pathways determined between deionized water and tertiary treated wastewater solutions

The oxidative reactions of bisphenol A (BPA) with radiolytically generated hydroxyl radicals were studied in both deionized water and tertiary treated wastewaters. In deionized water, bisphenol A reacts with the hydroxyl radical by addition to the aromatic ring, k = 6.9 × 10⁹ (±0.2) M⁻¹ s⁻¹, to eventually form the prominent, long-lived, hydroxylated intermediate product. In contrast, in tertiary treated water solutions, although the initial hydroxyl radical addition reaction occurs, the hydroxylation is averted, and a different mechanistic pathway ensues. The removal constant for the hydroxyl radical reaction with BPA is 0.45 ± 0.04 μmol/kGy, corresponding to an overall degradation efficiency of 76%.     

3-氨基-7,8-二甲氧基香豆素及其衍生物的合成及抗氧化活性

合成了3-氨基-7,8-二甲氧基香豆素及其衍生物共11个化合物,其中3个化合物(2b、2d、2e)为新型香豆素芳酰胺类化合物。 通过猝灭1,1-二苯基-2-三硝基苯肼(DPPH)、2,2'-联氮二(3-乙基苯并噻唑-6-磺酸)二铵盐阳离子自由基(ABTS^+·)和羟自由基实验考察了所合成化合物的抗氧化活性,结果表明化合物2b对DPPH自由基、羟自由基的清除能力超出或接近对照品维生素C,而衍生物2a、2b和2c的抗氧化活性优于母体。 故酰化可提高3-氨基-7,8-二甲氧基香豆素的抗氧化性能,尤其是普遍提高了羟基自由基的清除能力。     

SPECTROSCOPIC STUDIES OF CUTANEOUS PHOTOSENSITIZING AGENTS—X. A SPIN-TRAPPING AND DIRECT ELECTRON SPIN RESONANCE STUDY OF THE PHOTOCHEMICAL PATHWAYS OF DAUNOMYCIN AND ADRIAMYCIN

Abstract— Irradiation of daunomycin (or adriamycin) and the spin trap 5,5-dimethyl-l-pyrroline-1-oxide (DMPO) at 490 nm in the presence or in the absence of air generated the hydroxyl radical adduct (DMPO-OH). The observed DMPO-OH signal was not affected by the addition of hydroxyl radical scavengers (ethanol, formate), suggesting that direct trapping of the hydroxyl radical was not involved. The DMPO-OH signal was insensitive to superoxide dismutase and catalase, which ruled out the possibility of superoxide or H₂O₂ involvement. These findings demonstrate that daunomycin (or adriamycin) does not generate hydroxyl radicals or superoxide radical anions when subjected to 490-nm excitation. However, when daunomycin (or adriamycin) was irradiated at 310 nm DMPO adducts derived from two carbon-centered radicals, superoxide and the hydroxyl radical were detected. The superoxide adduct of DMPO was abolished by the addition of SOD, providing unequivocal evidence for the generation of the superoxide anion radical. The daunomycin semiquinone radical, observed upon 310-nm irradiation of daunomycin in the absence of DMPO, appears to be the precursor of the superoxide radical anion. One of the carbon-centered radicals trapped by DMPO exhibited a unique set of hyperfine parameters and was identified as an acyl radical. This suggests that the known photochemical deacylation of daunomycin occurs via a homolytic cleavage mechanism. The free radicals generated photolytically from adriamycin and daunomycin may be involved in the etiology of the skin ulceration and inflammation caused by these drugs. A knowledge of the dependence of these photogenerated radicals on the wavelength of excitation may be important in the development of adriamycin and daunomycin for photodynamic therapy.     

芳烃化合物液(均)相催化羟化反应研究进展

本综述以催化活性中心的原子种类为分类标准,着重于催化活性物种和催化反应机理的研究,系统地总结了自90年代以来苯及其芳烃化合物羟化合成在均相催化体系的研究进展.从研究趋势分析,人们从简单的无机盐均相催化体系过渡到合成仿生含氮配体的络合物作为催化剂的液-液两相催化体系,目前以铁、钒、铜和钯为催化活性中心的配合物液相催化体系的研究比较集中.从研究机理上来看,研究者运用多种研究手段和方法,提出了在各自研究体系中的*OH、高价金属氧配合物、金属氧或过氧或氢过氧化合物(或自由基)的羟化活性物种的自由基机理和配合物机理等.     

胆红素、铜离子与2,2′-联吡啶作用的光谱研究

基于2,2′-联吡啶芳杂环羟基化法,采用荧光光谱研究了胆红素与Cu2 的相互作用。探讨了胆红素与Cu2 反应生成羟自由基(.OH),2,2′-联吡啶发生芳杂环羟基化反应的过程和反应机理,产生的羟自由基导致无荧光的2,2′-联吡啶发生芳杂环羟基化,生成强荧光物质;加入自由基猝灭剂后,荧光强度的增强明显减弱。考察了试剂加入顺序、反应时间、酸度、Cu2 浓度、胆红素浓度,以及2,2′-联吡啶浓度等的影响。     

羟基自由基的流动注射荧光法测定

本文研究Fenton反应产生的·OH与Ce3+作用后Ce3+被氧化生成无荧光的Ce4+,通过测定Ce3+的荧光强度的下降可间接测定所产生的羟自由基,并结合流动注射技术,确定了体系最佳实验条件。同时测定抗氧化剂清除羟自由基的实验证明,该体系可作为在线筛选抗氧化剂以及在线测定羟自由基的方法。     

An in vitro study of the hydroxyl radical scavenging property of fluvastatin, and HMG-CoA reductase inhibitor.

We investigated the in vitro hydroxyl radical scavenging activity of fluvastatin, a 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitor. Fluvastatin showed hydroxyl radical scavenging activity as potent as that of dimethylthiourea and alpha-tocopherol, which are well-known respectively, as a hydroxyl radical scavenger and a natural antioxidant. Since this effect was not observed in other HMG-CoA reductase inhibitors, such as pravastatin and simvastatin, the scavenging effect of fluvastatin on hydroxyl radicals would not be a common property of HMG-CoA reductase inhibitors, but is derived from the unique chemical structure of fluvastatin. The hydroxyl radical scavenging activities of human metabolites of fluvastatin were also determined. All the tested metabolites possessing the fluorophenyl indole moiety showed activity. Among them, the metabolites which possess a phenolic hydroxyl group on the indole moiety showed stronger effects than that of fluvastatin. We suggest that the fluorophenyl indole moiety of fluvastatin is important for manifestation of the activity and that the phenolic hydroxyl group enhances the potency.     

Iron-chelating agents never suppress Fenton reaction but participate in quenching spin-trapped radicals

Hydroxyl radical formation by Fenton reaction in the presence of an iron-chelating agent such as EDTA was traced by two different assay methods; an electron spin resonance (ESR) spin-trapping method with 5,5-dimethyl-1-pyrroline N-oxide (DMPO), and high Performance liquid chromatography (HPLC)-fluorescence detection with terephthalic acid (TPA), a fluorescent probe for hydroxyl radicals. From the ESR spin-trapping measurement, it was observed that EDTA seemed to suppress hydroxyl radical formation with the increase of its concentration. On the other hand, hydroxyl radical formation by Fenton reaction was not affected by EDTA monitored by HPLC assay. Similar inconsistent effects of other iron-chelating agents such as nitrylotriacetic acid (NTA), diethylenetriamine penta acetic acid (DTPA), oxalate and citrate were also observed. On the addition of EDTA solution to the reaction mixture 10 min after the Fenton reaction started, when hydroxyl radical formation should have almost ceased but the ESR signal of DMPO-OH radicals could be detected, it was observed that the DMPO-OH signal disappeared rapidly. With the simultaneous addition of Fe(II) solution and EDTA after the Fenton reaction ceased, the DMPO-OH signal disappeared more rapidly. The results indicated that these chelating agents should enhance the quenching of [DMPO-OH] radicals by Fe(II), but they did not suppress Fenton reaction by forming chelates with iron ions.     

Determination of Reactive Intermediates During Anodic Oxygen-transfer Reactions at Lead Dioxide

IntroductionSincethepastdecade ,electrochemicalmethodsforelectrosynthesis ,wastewatertreatmentandorgan iccompounddeterminationhavebeenwidelydevel oped[1— 6 ] .Thebasicprincipleisthatorganicsub stancesareelectrochemicallyoxidizedintothedesiredproducts.However ,inmanycasesthedirectoxida tionoforganicspeciesinanaqueoussolutionatasim plemetaloracarbonelectrodeisnotpossiblebecausethehighpotentialsrequiredfortheoxidationoforgan iccompoundsusuallyresultinoxygenevolution .Therefore ,theproperchoiceof…     

Investigation of the anthracene-nitroxide hybrid molecule as a probe for hydroxyl radicals.

A new method for the determination of hydroxyl radicals is proposed. The method is based on the use of a hybrid molecule consisting of a fluorescent chromophore, anthracene, and a nitroxide radical. In the hybrid molecule, the nitroxide quenches the fluorescence of anthracene strongly. The reaction of hydroxyl radicals with dimethyl sulfoxide generates quantitatively methyl radicals, which then combine with the nitroxide moiety of the hybrid molecules to result in an increase in the fluorescence intensity. The fluorescence increase is proportional to the concentration of hydroxyl radicals. The proposed method is capable of detecting hydroxyl radicals generated in the Fenton system. It is a simple and sensitive technique for the determination of hydroxyl radicals.     

The "somersault" mechanism for the p-450 hydroxylation of hydrocarbons. The intervention of transient inverted metastable hydroperoxides

A series of model theoretical calculations are described that suggest a new mechanism for the oxidation step in enzymatic cytochrome P450 hydroxylation of saturated hydrocarbons. A new class of metastable metal hydroperoxides is described that involves the rearrangement of the ground-state metal hydroperoxide to its inverted isomeric form with a hydroxyl radical hydrogen bonded to the metal oxide (MO-OH --> MO....HO). The activation energy for this somersault motion of the FeO-OH group is 20.3 kcal/mol for the P450 model porphyrin iron(III) hydroperoxide [Por(SH)Fe(III)-OOH(-)] to produce the isomeric ferryl oxygen hydrogen bonded to an *OH radical [Por(SH)Fe(III)-O....HO(-)]. This isomeric metastable hydroperoxide, the proposed primary oxidant in the P450 hydroxylation reaction, is calculated to be 17.8 kcal/mol higher in energy than the ground-state iron(III) hydroperoxide Cpd 0. The first step of the proposed mechanism for isobutane oxidation is abstraction of a hydrogen atom from the C-H bond of isobutane by the hydrogen-bonded hydroxyl radical to produce a water molecule strongly hydrogen bonded to anionic Cpd II. The hydroxylation step involves a concerted but nonsynchronous transfer of a hydrogen atom from this newly formed, bound, water molecule to the ferryl oxygen with a concomitant rebound of the incipient *OH radical to the carbon radical of isobutane to produce the C-O bond of the final product, tert-butyl alcohol. The TS for the oxygen rebound step is 2 kcal/mol lower in energy than the hydrogen abstraction TS (DeltaE() = 19.5 kcal/mol). The overall proposed new mechanism is consistent with a lot of the ancillary experimental data for this enzymatic hydroxylation reaction.