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锰单原子纳米酶比色法检测碱性磷酸酶活性

王波 ,  张续东 ,  康革 ,  刘芳宁 ,  赵丹 ,  陈传霞 ,  逯一中

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引用本文: 王波, 张续东, 康革, 刘芳宁, 赵丹, 陈传霞, 逯一中. 锰单原子纳米酶比色法检测碱性磷酸酶活性[J]. 分析化学, 2022, 50(1): 54-63. doi: 10.19756/j.issn.0253-3820.210812 shu
Citation:  WANG Bo,  ZHANG Xu-Dong,  KANG Ge,  LIU Fang-Ning,  ZHAO Dan,  CHEN Chuan-Xia,  LU Yi-Zhong. Colorimetric Detection of Alkaline Phosphatase Activity Based on Manganese Single Atom Nanozyme[J]. Chinese Journal of Analytical Chemistry, 2022, 50(1): 54-63. doi: 10.19756/j.issn.0253-3820.210812 shu

锰单原子纳米酶比色法检测碱性磷酸酶活性

  • 1.

    济南大学材料科学与工程学院, 济南 250022;

  • 2.

    洛阳理工学院环境工程与化学学院, 洛阳 471023

    • 通讯作者: 赵丹,E-mail:lzdzhaodan@163.com; 陈传霞,E-mail:mse_chencx@ujn.edu.cn
  • 基金项目:

    国家自然科学基金项目（Nos.21902061，22104046，21904048，22172063）和山东省自然科学基金项目（Nos.ZR2020QB033，ZR2019YQ10）资助。

摘要: 利用溶剂热法，合成了具有类过氧化物酶活性的锰单原子纳米酶，用于碱性磷酸酶（ALP）活性的检测。以甲酰胺为碳源和氮源，MnCl2为锰源，经180℃反应12 h，获得富含单分散Mn-N4位点的锰、氮共掺杂碳材料（f-MnNC）。通过透射电子显微镜、X-射线衍射仪、X-射线光电子能谱、红外光谱等对f-MnNC的结构和形貌进行了表征，利用紫外-可见吸收光谱对f-MnNC的类过氧化物酶活性进行了研究。具有类过氧化物酶活性的f-MnNC可催化H2O2将无色的3，3'，5，5'-四甲基联苯胺（TMB）氧化为蓝色的氧化态TMB （oxTMB）。考察了f-MnNC催化TMB-H2O2显色反应条件对此过氧化物酶活性的影响，并进行了稳态动力学分析。结果表明，f-MnNC的酶催化反应遵循典型的Michaelis-Menten动力学模型。利用抗坏血酸磷酸酯（AA2P）为底物，ALP催化AA2P水解产生抗坏血酸（AA），AA具有还原性，可通过还原oxTMB使得体系蓝色变浅，吸光度降低。信号降低程度与ALP活性有关，据此可对ALP活性进行定量检测。本方法检测ALP活性的线性范围为0.1~10 mU/mL，检出限（S/N=3）为0.059 mU/mL。其它生物体系中常见蛋白和生物酶对ALP检测无明显干扰，表明本方法检测ALP活性具有良好的选择性。将本方法用于血清中ALP活性的检测，结果令人满意。

English


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  • 收稿日期:  2021-10-26
  • 修回日期:  2021-11-24
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