College of Chemistry and Material Science, Shandong Agricultural University, Daizong Street 61, Taian 271018, Shandong, China.
J Hazard Mater. 2011 Jun 15;190(1-3):480-5. doi: 10.1016/j.jhazmat.2011.03.071. Epub 2011 Mar 29.
A new electrochemical biosensor for directly detecting DNA damage induced by acrylamide (AA) and its metabolite was presented in this work. The graphene-ionic liquid-Nafion modified pyrolytic graphite electrode (PGE) was prepared, and then horseradish peroxidase (HRP) and natural double-stranded DNA were alternately assembled on the modified electrode by the layer-by-layer method. The PGE/graphene-ionic liquid-Nafion and the construction of the (HRP/DNA)(n) film were characterized by electrochemical impedance spectroscopy. With the guanine signal in DNA as an indicator, the damage of DNA was detected by differential pulse voltammetry after PGE/graphene-ionic liquid-Nafion/(HRP/DNA)(n) was incubated in AA solution or AA+H(2)O(2) solution at 37°C. This method provides a new model to mimic and directly detect DNA damage induced by chemical pollutants and their metabolites in vitro. The results indicated that, in the presence of H(2)O(2), HRP was activated and catalyzed the transformation of AA to glycidamide, which could form DNA adducts and induce more serious damage of DNA than AA. In order to further verify these results, UV-vis spectrophotometry was also used to investigate DNA damage induced by AA and its metabolites in solution and the similar results were obtained.
本文提出了一种新的电化学生物传感器，用于直接检测丙烯酰胺（AA）及其代谢物诱导的 DNA 损伤。制备了石墨烯-离子液体-聚全氟磺酸膜修饰的热解石墨电极（PGE），然后通过层层自组装法将辣根过氧化物酶（HRP）和天然双链 DNA 交替组装在修饰电极上。通过电化学阻抗谱对 PGE/石墨烯-离子液体-聚全氟磺酸膜和（HRP/DNA）（n）膜的构建进行了表征。用 DNA 中的鸟嘌呤信号作为指示剂，在 PGE/石墨烯-离子液体-聚全氟磺酸膜/（HRP/DNA）（n）孵育于 AA 溶液或 AA+H2O2 溶液中 37°C 后，通过差分脉冲伏安法检测 DNA 的损伤。该方法提供了一种新的模型，可用于体外模拟和直接检测化学污染物及其代谢物诱导的 DNA 损伤。结果表明，在 H2O2 的存在下，HRP 被激活并催化 AA 转化为缩水甘油酰胺，这可能形成 DNA 加合物，并导致比 AA 更严重的 DNA 损伤。为了进一步验证这些结果，还使用紫外可见分光光度法研究了溶液中 AA 及其代谢物诱导的 DNA 损伤，得到了相似的结果。
