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检测和分析反应性氧物种 (ROS):缓冲组件不是旁观者

Shubham Bansal¹, Muskan Gori¹, Joanna Afokai Quaye¹

¹Department of Chemistry and Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, Georgia 30301 United States.

Analytical chemistry

|June 3, 2025

概括

像HEPES和Tris这样的常见有机缓冲器通过消耗ROS和与探针相互作用来干扰反应性氧物种 (ROS) 检测. 这导致ROS度测量不准确和假负结果,需要在实验中谨慎选择缓冲器.

科学领域:

生物化学生物化学
细胞生物学细胞生物学
分析化学分析化学

背景情况:

活性氧物种 (ROS) 在生物过程中至关重要,引发了对其准确测量的兴趣.
现有文献显示ROS度数据的显著变化,阻碍了可比性.
一个影响实验结果的关键因素,即缓冲元件与ROS的反应性,经常被忽视.

研究的目的:

为了研究常见的有机缓冲器与ROS的化学反应性.
确定缓冲器-ROS相互作用如何影响ROS度测量的准确性.
为可靠的ROS检测提供选择适当缓冲器的指导.

主要方法:

测试了HEPES,Tris,MES,酸盐,酸和PBS的反应性,使用低酸盐 (NaOCl) 和过氧化 (H2O2) 探针.
评估了缓冲元件与用于H2O2和过氧化 (ONOO-) 检测的酸盐基探针的相互作用.
量化了缓冲分子的ROS消耗率.

主要成果:

大多数测试的有机缓冲器迅速消耗NaOCl,导致ROS检测的错误负结果.
缓冲元件干扰酸盐基探头,导致H2O2和ONOO-的测量不准确.
低至20毫米的HEPES,MES,酸和Tris度可以在1秒内消耗1毫米的化物.

结论:

由于化学干扰,通常使用的有机缓冲器不适合用于ROS度的确定.
建议避免这些缓冲区,以防止实验结果和解释中的重大错误.
研究人员必须考虑缓冲化学,以提高ROS检测实验的严谨性和可靠性.

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