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Citation: KONG Hao, ZHANG Qiang, ZHANG Wei, LIU Weiwen, CAO Chengxi, FAN Liuyin. Visualized and quantitative detection of horseradish peroxidase based on redox reaction boundary[J]. Chinese Journal of Chromatography, ;2020, 38(2): 177-182. doi: 10.3724/SP.J.1123.2019.04030 shu

Visualized and quantitative detection of horseradish peroxidase based on redox reaction boundary

KONG Hao ^1,2 ,
ZHANG Qiang ^2,3 ,
ZHANG Wei ² ,
LIU Weiwen ³ ,
CAO Chengxi ^3,, ,
FAN Liuyin ^1,,

1.
Student Innovation Center, Shanghai Jiao Tong University, Shanghai 200240, China;
2.
School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;
3.
School of Electronic Information & Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China

Corresponding author: CAO Chengxi, FAN Liuyin,
Received Date: 16 April 2019

Fund Project: National Natural Science Foundation of China (Nos. 21675067, 31727801, 21605101)

A new portable method for the detection of horseradish peroxidase (HRP) is reported in this paper. Visualized and quantitative detection of HRP was achieved based on the theory of electrophoresis titration (ET) and redox reaction boundary (RB). The ET-RB model was built on the 3,3',5,5'-tetramethyl benzidine chromogenic system because it significantly increases the efficiency of the chromogenic reaction and can react with L-ascorbic acid. A series of experiments were performed on a tiny and portable titration chip based on the material of polymethyl methacrylate designed for the developed model. The composition of the titration gel was optimized to ensure higher detection sensitivity and stable detection performance. The experimental results manifested a log-linear relationship between the moving distance of the RB and the HRP concentration. The dynamic range of the developed method ranged from 0.002 to 0.073 mg/L, and the detection could be accomplished within 10 min. Visualized and quantitative detection could be realized by reading the moving distance of the colored boundary with the naked eyes instead of using any signal-reading device or analyzing instrument. Thus, this method has great potential for further application to the development of a real-time detection method for various target substances based on the detection of peroxidase activity.
- redox reaction,
- electrophoresis titration (ET),
- real-time detection,
- visualization,
- horseradish peroxidase (HRP)
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