【摘要】 研究产生和消除过氧亚硝基(Peroxynitriteanion,ONOO?)是自由基生物学和医学的一项紧迫任务。为此,建立了一种新的产生和检测ONOO?的化学发光体系,并评估了检测的最佳条件。该方法为:10mmol/L羟胺与0.5mol/LNaOH、1mmol/L、乙二胺四乙酸二钠(Disodiumethylenediaminetetraacetate,EDTA)反应产生ONOO?,随后用MnO2粉末过滤反应液以除去H2O2,然后在测量管内加入100μL抗氧化剂和860μL0.1mol/L的碳酸盐缓冲液(pH10),再加入40μL过滤后的ONOO?反应液、混匀、延迟10s,测10s化学发光强度。以抗氧化剂茶多酚、抗坏血酸、芦丁和黄芩苷作清除ONOO?的实验。结果表明,本体系是一种灵敏价廉、可靠易行的筛选ONOO?清除剂的新方法。更多还原

【Abstract】 The study for generating and scavenging peroxynitrite anions (ONOO?) is an urgent task in free radi-cal biology and medicine. Therefore, a novel chemiluminescence system for producing and measuring ONOO? has been established. The optimal conditions for detecting ONOO? were evaluated. The method can be described as fol-lows. A mixture solution containing 10 mmo l / L hydroxylamine, 0. 5 mo l / L NaOH and 1mmo l / L EDTA was stirred vigorously in aerobic conditions for about 3 h to form ONOO?. Certain amount of MnO2 powder was added to the mixture solution to eliminate H2O2 generated in the reaction. Then 10 0 μL antioxidant and 860μL 0.1mo l / L carbon-ate buffer (pH10) were added into a test tube. By adding 40μL filtrated ONOO? solution into the tube, chemilumines-cence (CL) reaction was initiated. Ten seconds after the solution was uniformly mixed, CL intensity of the reaction solution could be determined in a measurement of 10 seconds. The method was tested with tea polyphenol, ascorbic acid, rutin and baicalin, whish are known to have effects of scavenging ONOO?. The tests show that the system is sensitive, low?cost, credible and easy method for screening ONOO?.更多还原