The Amplex Red/horseradish peroxidase (AR/HRP) assay for H₂O₂ is one of the most sensitive and simple approaches for H₂O₂ quantification, which is effected by measuring the highly-fluorescent resorufin formed from oxidation of AR by the oxidizing intermediates generated by reaction of HRP and H₂O₂. The direct reactions of S(−II) with both H₂O₂ and resorufin are too slow to be of relevance on analytical timescales, however, the reaction between S(−II) and the HRP/H₂O₂ oxidizing intermediates is rapid enough to compete with the desired reaction of these oxidizing intermediates with AR, suppressing formation of the resorufin analyte. As this mode of interference can be considered simply a competition between the AR reagent and S(−II) for the intermediate oxidizing species, a simple equation is derived in this work enabling one to correct for this interference and obtain a good estimate of the true H₂O₂ concentration after measuring the apparent H₂O₂ concentration and the S(−II) concentration. This mode of interference is general to any compound that can act as a HRP substrate even if not directly reactive with H₂O₂. As such, the approach described is widely applicable to many potential reducing interferents and opens up the use of the AR/HRP assay to a much wider range of conditions, as well as demonstrating the utility of explicitly considering the mechanism of the analytical process.

中文翻译：

---

使用Amplex Red /辣根过氧化物酶测定法定量测定在硫化物存在下的痕量过氧化氢

H ₂ O ₂的Amplex Red /辣根过氧化物酶（AR / HRP）测定是H ₂ O ₂定量最敏感，最简单的方法之一，该方法是通过测量由AR氧化而形成的高荧光试卤灵来实现的。 HRP和H ₂ O ₂反应生成的氧化中间体。S（-II）与H ₂ O ₂和试卤灵的直接反应太慢，无法与分析时标相关，但是S（-II）与HRP / H ₂ O ₂之间的反应氧化中间体的速度足够快，足以与这些氧化中间体与AR进行所需的反应，从而抑制了间苯二酚分析物的形成。由于这种干扰方式可以简单地认为是AR试剂与S（-II）之间对中间氧化物质的竞争，因此在此工作中得出了一个简单的方程式，使人们可以纠正这种干扰并获得对真实干扰的良好估计。 ħ ₂ Ó ₂测量表观之后轰浓度₂ ö ₂浓度和S（-II）的浓度。这种干扰模式是所有可以用作HRP底物的化合物所通用的，即使它们不会直接与H ₂ O ₂反应。这样，所描述的方法可广泛地应用于许多潜在的减少干扰物，并且将AR / HRP分析的使用扩展到更广泛的条件范围，并且证明了明确考虑分析过程机理的实用性。