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Updated: Aug 13, 2026
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Chemiluminescence-based Assays for Detection of Nitric Oxide and its Derivatives from Autoxidation and Nitrosated Compounds
Published on: February 16, 2022
Sequential injection analysis with chemiluminescence detection for the antioxidative activity against singlet oxygen
Aoi Miyamoto1, Kuniko Nakamura, Yoshihito Ohba
1Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Summary
A new sequential injection analysis (SIA) method with chemiluminescence (CL) detection measures antioxidative activity against singlet oxygen ((1)O2). This rapid and reproducible system efficiently screens compounds for their antioxidant properties.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Chemical Analysis
Background:
- Singlet oxygen ((1)O2) is a reactive oxygen species implicated in oxidative stress.
- Measuring antioxidant activity against (1)O2 is crucial for understanding cellular damage and developing protective agents.
- Existing methods for assessing antioxidative activity can be time-consuming and reagent-intensive.
Purpose of the Study:
- To develop a rapid and reproducible analytical method for quantifying antioxidative activity against singlet oxygen ((1)O2).
- To optimize a sequential injection analysis (SIA) system coupled with chemiluminescence (CL) detection for this purpose.
- To validate the system's performance using known antioxidants and assess its applicability to complex samples.
Main Methods:
- A sequential injection analysis (SIA) system was designed utilizing a luminol-based chemiluminescence (CL) reaction.
- Singlet oxygen ((1)O2) was generated using a lactoperoxidase-hydrogen peroxide-bromide ion system.
- Antioxidative activity was determined by measuring the CL signal attenuation caused by the quenching of (1)O2 by antioxidants.
Main Results:
- The SIA-CL system was optimized for carrier solution (100 mM sodium acetate buffer, pH 4.5), flow rate, reagent concentrations, and aspiration order.
- Excellent reproducibility was achieved, with relative standard deviations for antioxidative activity measurements below 1.6% for both within-day and between-day analyses (using 20 µM Trolox).
- The system demonstrated high throughput, analyzing 10 samples, including vitamin supplements, within 15 minutes, with minimal sample and reagent consumption.
Conclusions:
- The developed SIA-CL method provides a rapid, reproducible, and efficient approach for measuring antioxidative activity against singlet oxygen ((1)O2).
- This method is suitable for the high-throughput screening of compounds and natural products for their ability to neutralize singlet oxygen.
- The system's minimal consumption of reagents and samples makes it a cost-effective tool for antioxidant research.
