Analyzing the SOD-like activity of nanozymes in vitro is of great importance for identifying new nanozymes and predicting their potential biological effects in vivo. However, false negative or positive results occasionally occur due to the mismatch between the detection methods and nanozymes. Here, five typical SOD-like nanozymes, including CeO₂, Mn₃O₄, Prussian blue (PB), PCN222-Mn, and Pt NPs, have been used to evaluate the sensitivity and accuracy of several commonly used in vitro detection methods. By systematically analyzing the detection results, several precautions have been taken. (1) The hydroethidine (HE) probe could be disturbed by the nanozyme with oxidative ability. (2) The nitro blue tetrazolium (NBT) probe has a moderate sensitivity due to the poor water solubility of its reduced product. (3) The water-soluble tetrazolium salt (WST)-8 probe has a higher sensitivity than both NBT and iodonitrotetrazolium chloride (INT). (4) The detection system using the irradiation of riboflavin to produce ˙O₂⁻ might be interfered by the nanozyme with photosensibility. (5) Both the quality of DMPO and incubation time are important factors for electron paramagnetic resonance (EPR) measurement. This study will be useful for choosing more suitable in vitro detection methods of SOD-like activity for nanozymes in the future.

中文翻译：

---

体外测量超氧化物歧化酶样纳米酶的活性：一项比较研究

在体外分析纳米酶的类SOD活性对于鉴定新的纳米酶并预测其在体内的潜在生物学作用非常重要。但是，由于检测方法和纳米酶之间的不匹配，有时会出现假阴性或阳性结果。在这里，五个典型的类似于SOD的纳米酶，包括CeO ₂，Mn ₃ O ₄，普鲁士蓝（PB），PCN222-Mn和Pt NPs，已用于评估几种常用的体外敏感性和准确性。检测方法。通过系统地分析检测结果，已经采取了几种预防措施。（1）带有氧化能力的纳米酶可能会干扰氢乙啶（HE）探针。（2）硝基蓝四唑（NBT）探针由于还原产物的水溶性差而具有中等灵敏度。（3）水溶性四唑盐（WST）-8探针比NBT和氯化碘硝基四唑鎓（INT）都具有更高的灵敏度。（4）使用核黄素生产邻照射检测系统₂ ^-可能通过与photosensibility的nanozyme干扰。（5）DMPO的质量和孵育时间都是电子顺磁共振（EPR）测量的重要因素。这项研究将有助于选择更适合的体外 纳米酶的类SOD活性检测方法。