Abstract Hydrogen peroxide has important roles within cellular functions, as a prevalent form of Reactive Oxygen Species, detection within mammalian cells is of metabolic importance; typically requiring cell lysis or fluorescence-based methods to quantify. Herein, we explore the novel use of Prussian blue mediated, pad printed carbon electrodes to allow the indirect detection of cellular peroxides in bulk culture media, which facilitates non-invasive, real-time detection. Electrodes demonstrated capacity to detect H2O2 with a linear range of 1-200 μM in CMEM (R2 = 0.9988), enabling detection of peroxides found in culture media and lysate. Developed electrodes had a Limit of Detection (LOD) of 0.41 μM H2O2 in Britton-Robinson Buffer (BRB), 0.38 μM in Eagle's Minimum Essential Medium (EMEM) and 9.19 μM in Dulbecco's Modified Eagles Medium (DMEM). Electrodes were tested in a conventional 5% serum supplemented EMEM (CMEM) and demonstrated an LOD of 0.5 μM and LOQ of 0.9 μM. The results demonstrate proof of concept for monitoring H2O2 in complex culture media with potential long-term use and reusability using simple, pad printed Prussian Blue / Carbon electrodes. The lack of further modification, and cost-effectiveness of these disposable electrodes could offer great advancement to monitoring of peroxides in complex media.

中文翻译：

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用于细胞培养中实时过氧化物传感的移印普鲁士蓝掺杂碳墨水

摘要 过氧化氢在细胞功能中具有重要作用，作为活性氧的一种普遍形式，在哺乳动物细胞内检测具有代谢重要性；通常需要细胞裂解或基于荧光的方法来量化。在此，我们探索了普鲁士蓝介导的移印碳电极的新用途，以允许间接检测散装培养基中的细胞过氧化物，这有助于非侵入性的实时检测。电极在 CMEM (R2 = 0.9988) 中以 1-200 μM 的线性范围显示出检测 H2O2 的能力，从而能够检测培养基和裂解物中发现的过氧化物。开发的电极在 Britton-Robinson 缓冲液 (BRB) 中的检测限 (LOD) 为 0.41 μM H2O2，在 Eagle 最低必需培养基 (EMEM) 中为 0.38 μM，在 Dulbecco' 中为 9.19 μM s 改良 Eagles 培养基 (DMEM)。电极在传统的 5% 血清补充 EMEM (CMEM) 中进行测试，结果显示 LOD 为 0.5 μM，LOQ 为 0.9 μM。结果证明了使用简单的移印普鲁士蓝/碳电极监测复杂培养基中 H2O2 的概念证明，具有潜在的长期使用和可重复使用性。这些一次性电极无需进一步修改，成本效益高，可以为监测复杂介质中的过氧化物提供巨大进步。