基于铁基γ-环糊精纳米粒子的类过氧化物酶活性比色检测肌氨酸  

Colorimetric Detection of Sarcosine Based on Peroxidase-like Activity of Fe-basedγ-Cyclodextrin Nanoparticles

作　　者：王皓 王婷婷 宋浩 刘芳宁 陈传霞 逯一中 WANG Hao;WANG Ting-Ting;SONG Hao;LIU Fang-Ning;CHEN Chuan-Xia;LU Yi-Zhong(School of Materials Science and Engineering,University of Jinan,Jinan 250022,China)

机构地区：[1]济南大学材料科学与工程学院,济南250022

出　　处：《分析化学》2024年第8期1072-1081,共10页Chinese Journal of Analytical Chemistry

基　　金：山东省自然科学基金项目(No.ZR2023MB019);国家自然科学基金项目(No.22172063);电分析化学国家重点实验室开放课题项目(No.SKLEAC202314)资助。

摘　　要：采用溶剂热法合成了具有类过氧化物酶活性的铁基γ-环糊精纳米粒子(Fe-γ-CD),并基于此构建了简单、灵敏测定肌氨酸(SAR)的比色传感方法。Fe-γ-CD可催化过氧化氢(H_(2)O_(2))氧化无色的3,3′,5,5′-四甲基联苯胺(TMB),生成蓝色的氧化态TMB(oxTMB),在652 nm处有强吸收峰。考察了反应条件对Fe-γ-CD催化的TMB-H_(2)O_(2)显色反应的影响,并进行了催化机理和稳态动力学分析。实验结果表明,Fe-γ-CD催化H_(2)O_(2)产生羟基自由基、单线态氧和超氧自由基,这些活性氧物种进一步氧化TMB,此催化反应历程遵循典型的Michaelis-Menten动力学模型。SAR在肌氨酸氧化酶(SO_(x))催化下可水解产生H_(2)O_(2),具有类过氧化物酶活性的Fe-γ-CD催化H_(2)O_(2)氧化TMB,使得溶液颜色变蓝,并且在652 nm处的吸光度增加,吸光度升高程度与SAR的浓度有关,由此可对SAR浓度进行定量检测。本方法的线性检测范围为1.0~70.0μmol/L,检出限为0.46μmol/L(S/N=3),并具有良好的特异性。将本方法用于血清中SAR含量的测定,结果令人满意。Fe-basedγ-cyclodextrin nanoparticles(Fe-γ-CD)with peroxidase-like activity were successfully synthesized through a solvothermal approach for establishing a simple and sensitive sarcosine(SAR)assay method.Fe-γ-CD could catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine(TMB)to blue oxidized TMB(oxTMB)in the presence of H_(2)O_(2),accompanied by a characteristic absorption peak centered at 652 nm.The effects of reaction conditions were investigated,and the catalytic mechanism as well as the steady-state kinetics were analyzed.Fe-γ-CD could catalyze H_(2)O_(2)to generate hydroxyl radical,singlet oxygen and superoxide radical,these reactive oxygen species with robust oxidizability further oxidized TMB.This process followed a typical Michaelis-Menten kinetic model.With the assistance of sarcosine oxidase(SO_(x)),SAR was hydrolyzed into H_(2)O_(2)to trigger Fe-γ-CD catalyzed chromogenic reaction,resulting in deepened color and increased absorbance.The degree of colorimetric signal change was related to SAR concentration and thus SAR could be quantitatively detected.The linear range and detection limit were 1.0–70.0μmol/L and 0.46μmol/L,respectively.Typical amino acids and metal ions had no obvious interference on SAR detection,indicating a good selectivity.The method was successfully applied to determination of SAR level in serum with satisfactory results.

关 键 词：铁基γ-环糊精纳米粒子 肌氨酸 过氧化氢 比色 

分 类 号：R446.1[医药卫生—诊断学] O657.3[医药卫生—临床医学]