高等学校化学学报

高等学校化学学报 ›› 2012, Vol. 33 ›› Issue (01): 54.doi: 10.3969/j.issn.0251-0790.2012.01.009

• 分析化学 • 上一篇    下一篇

一种基于多羧基荧光共轭聚电解质的氨基酸检测新方法

范宏亮1,2, 张涛1, 李祖宏1, 吴丽媛3, 金钦汉1   

  1. 1. 浙江大学工业控制技术国家重点实验室, 智能系统与控制研究所, 分析仪器研究中心,杭州 310058;
    2. 浙江大学生命科学学院, 杭州 310058;
    3. 吉林大学分子酶学工程教育部重点实验室, 长春 130021
  • 收稿日期:2011-01-06 出版日期:2012-01-10 发布日期:2011-12-20
  • 通讯作者: 张涛,男,博士,副教授,主要从事生化传感方面的研究.E-mail:zhtao@zju.edu.cn;金钦汉,男,教授,博士生导师,主要从事光谱分析与分析仪器方面的研究.E-mail:qhjin@zju.edu.cn E-mail:zhtao@zju.edu.cn; qhjin@zju.edu.cn
  • 基金资助:

    国家自然科学基金(批准号: 20805042)、 浙江省自然科学基金(批准号: Y4090154)、 中央高校基本科研业务费专项资金(批准号: 2009QNA5006)和科技部创新方法专项(批准号: 2008IM040800)资助.

Specific Assay of Amino Acids Based on a Fluorescent Conjugated Polyelectrolyte with Multi-carboxylates

FAN Hong-Liang1,2, ZHANG Tao1, LI Zu-Hong1, WU Li-Yuan3, JIN Qin-Han1   

  1. 1. Research Center for Analytical Instrumentation, Institute of Cyber-Systems and Control, State Key Laboratory of Industrial Control Technology,Zhejiang University, Hangzhou 310058, China;
    2. College of Life Science, Zhejiang University, Hangzhou 310058, China;
    3. Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, Jilin University, Changchun 130021, China
  • Received:2011-01-06 Online:2012-01-10 Published:2011-12-20
  • Contact: ZHANG Tao E-mail:zhtao@zju.edu.cn; qhjin@zju.edu.cn

PDF (PC)

被引次数

1

摘要： 基于多羧基荧光共轭聚电解质PPE-(COOK)4, 发展了一种检测半胱氨酸(Cys)和组氨酸(His)的简便方法. 该方法以PPE-(COOK)4与Cu2+所构成的猝灭体系为基础, 当体系中存在半胱氨酸或组氨酸时, PPE-(COOK)4/Cu2+体系的荧光恢复, 从而实现对上述氨基酸的测定. 由于PPE-(COOK)4具有类似四乙酸乙二胺(EDTA)的螯状侧链结构, 使其在磷酸盐缓冲液(PBS, 10 mmol/L, pH=7.4)中对Cu2+具有超高的灵敏度和选择性, 因此可以借助各种金属掩蔽离子来提高氨基酸检测的特异性. 实验结果表明, 在PPE-(COOK)4/Cu2+体系中引入Ni2+或Hg2+后, 可分别实现对半胱氨酸和组氨酸的特异性检测, 线性范围分别为1~5和0.5~2.5 μmol/L, 检出限分别为0.15和0.04 μmol/L.

关键词: 共轭聚电解质, 生化传感器, 铜离子探针, 氨基酸

Abstract: A simple assay for cysteine(Cys) and histidine(His) based on a conjugated polyelectrolyte with multi-carboxylate groups was developed. The proposed approach takes advantage of the emission quenching of PPE-(COOK)4 caused by Cu2+, which can be efficiently restored in the presence of Cys or His. Thus, the rapid assay of both amino acids can be carried out by measuring the fluorescence recovery. With the high similarity between the polymer’s side groups and EDTA, PPE-(COOK)4 in phosphate buffer solution exhibits not only high sensitivity but remarkable specificity towards Cu2+, which makes it possible to introduce various shielding metals for improving the selectivity of Cys and His assays. The results demonstrated that both Cys and His can be assayed sensitively and selectively by adding Ni2+ or Hg2+ to PPE-(COOK)4/Cu2+ system. The limit of detection is 0.15 μmol/L for Cys and 0.04 μmol/L for His, with the linear range of 1-5 μmol/L and 0.5-2.5 μmol/L, respectively.

Key words: Conjugated polyelectrolyte, Biosensor, Cu2+ probe, Amino acid

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