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Development of an antioxidant assay to study oxidative potential of airborne particulate matter

机译:抗氧化试验的发展,以研究空气传播颗粒物质的氧化潜力

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Oxidative potential is a measure of redox activity of airborne particulate matter (PM) and is often used as a surrogate to estimate one form of PM toxicity. The evaluation of oxidative potential in a physiologically relevant environment is always challenging. In this work, we developed a chromatographic method, employing an ultra-high-performance liquid chromatograph coupled to a triple–quadruple mass spectrometer, to determine the oxidative potential of PM from different sources. To this purpose, we measured the PM-induced oxidation of glutathione, cysteine, and ascorbic acid, and formation of glutathione disulfide and cystine, following PM addition to simulated epithelial lining fluids, which, in addition to the antioxidants, contained inorganic salts, a phospholipid, and proteins. The new method showed high precision and, when applied to standard reference PM, the oxidative potential was found to increase with the reaction time and PM concentration in the lung fluid. The antioxidant depletion rates were considerably higher than the rates found with the conventional dithiothreitol assay, indicating the higher sensitivity of the new method. The presence of the lung fluid inorganic species increased the oxidative potential determined through glutathione and cysteine, but showed an opposite effect with ascorbic acid, whereas the presence of proteins resulted in a moderate decrease in the oxidative potential. In the presence of PM2.5, glutathione and cysteine demonstrated similar depletion patterns, which were noticeably different from that of ascorbic acid, suggesting that cysteine could be used as an alternative to glutathione for probing oxidative potential.
机译:氧化潜力是空气传播颗粒物质(PM)的氧化还原活性的量度,并且通常用作替代替代以估计一种形式的PM毒性。在生理相关环境中对氧化潜力的评估总是具有挑战性的。在这项工作中,我们开发了一种色谱法,采用超高性能液相色谱仪耦合到三倍质谱仪,以确定来自不同来源的PM的氧化潜力。为此目的,我们测量了PM诱导的谷胱甘肽,半胱氨酸和抗坏血酸的氧化,以及在模拟上皮衬里的PM加入后形成谷胱甘肽二硫化物和胱氨酸,除了抗氧化剂,含有无机盐,a磷脂和蛋白质。新方法显示出高精度,并且当施加到标准参考PM时,发现氧化电位随肺液中的反应时间和PM浓度而增加。抗氧化剂耗竭率大于常规二硫醇测定的速率高得多,表明新方法的敏感性较高。肺液无机物种的存在增加了通过谷胱甘肽和半胱氨酸测定的氧化潜力,但与抗坏血酸表现出相反的效果,而蛋白质的存在导致氧化潜力的中等降低。在PM2.5的存在下,谷胱甘肽和半胱氨酸展示了类似的耗竭模式,其明显不同于抗坏血酸的缺陷模式,表明半胱氨酸可以用作抑制氧化潜力的谷胱甘肽的替代方法。

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