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Simultaneous quantification of peroxidase and ascorbic acid in biosamples with an automatic system based on a
Yang Zhao1, Qiao-Jing Li, Yong-Sheng Li
1School of Chemical Engineering, Sichuan University, Chengdu 610065, China. lysgxf2005@qq.com.
The Analyst
|June 24, 2020
Summary
A new flow injection analysis system simultaneously quantifies peroxidase (POD) and ascorbic acid (AsA) in plant samples. This method utilizes novel carbon dots as a POD simulant, offering accurate and efficient analysis.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Materials Science
Background:
- Peroxidase (POD) and ascorbic acid (AsA) are crucial antioxidants with dynamic concentrations in biological systems.
- Ascorbic acid (AsA) interferes with spectrophotometric assays of peroxidase (POD) activity due to an extinction reaction with chromogenic products.
- Accurate quantification of both POD and AsA is essential for understanding their synergistic protective roles against oxidative damage.
Purpose of the Study:
- To develop an automatic analysis system for simultaneous quantification of POD and AsA.
- To establish a novel simulative enzyme using carbon dots (sPOD-CDs) for POD activity assays.
- To investigate the role of ascorbate ions in AsA reducibility and validate the system in plant samples.
Main Methods:
- Development of a flow injection analysis (FIA) system leveraging chromogenic and extinction phenomena.
- Hydrothermal synthesis of methylthymol blue/FeCl3-based carbon dots (sPOD-CDs) as a POD simulant.
- Optimization of the FIA system for simultaneous determination of POD (73-440 U L⁻¹) and AsA (4-60 mg L⁻¹).
Main Results:
- The FIA system successfully quantified POD and AsA simultaneously with high sample throughput (40 samples/h) and repeatability (RSD ≤ 1.4%).
- Synthesized sPOD-CDs demonstrated no background absorption and correlated well with simulative POD activity, effectively replacing natural enzymes.
- Matrix interference was negligible in plant samples (recoveries 95%-105%), and results aligned with reference experiments (RSD ≤ 2.80%).
Conclusions:
- A robust FIA method for simultaneous POD and AsA determination was established using a novel carbon dot-based simulative enzyme.
- The system accurately quantifies POD and AsA in plant matrices, confirming the role of ascorbate ions in AsA's reducing capacity.
- This approach offers an efficient and reliable tool for biochemical analysis in biological and environmental samples.
