# Colorimetric detection of chloroperoxyl radical in reactive chlorine species solutions

**Authors:** Hiroyuki Kawata, Shunsuke Odai, Hisataka Goda, Yuki Okawa, Toshiaki Kamachi

PMC · DOI: 10.1371/journal.pone.0334046 · PLOS One · 2025-10-21

## TL;DR

Researchers developed a colorimetric method to detect and quantify the chloroperoxyl radical, a key disinfectant in chlorous acid water, which could improve understanding of its disinfecting properties.

## Contribution

The first colorimetric quantification method for the chloroperoxyl radical (ClOO•) is introduced, enabling selective and stable detection.

## Key findings

- ClOO• was found to be the only detectable oxidant in the solution with a stability exceeding 100 hours.
- The DPD-based colorimetric method showed a wide detection range and excellent linearity for quantifying ClOO•.
- Cation exchange successfully produced a high-purity ClOO• solution confirmed by ESR and chlorine concentration measurements.

## Abstract

Chlorous acid water (CAW) is a chlorine-based disinfectant approved as a food additive in Japan. CAW is synthesized by the reaction of chloric acid aqueous solution with hydrogen peroxide under acidic conditions. However, in this synthesis method, various ions—such as Na+ from NaClO3 and the conjugate base of the acid used—remain in the solution, hindering the selective detection and decreasing stability of chloroperoxyl radical (ClOO•), a potential key disinfectant species. In this study, we aimed to establish a colorimetric quantification method for ClOO•. We prepared a high-purity ClOO• solution (ClO2_cx) by cation exchange and its purity and stability were evaluated using electron spin resonance (ESR) spectroscopy and total chlorine concentration measurements. Furthermore, several colorimetric methods—including DPD, TMB, and DPPH assays—were examined to quantify ClOO•, and their sensitivity and selectivity were comparatively assessed. ClOO• was the sole detectable oxidant in the solution, with a lifetime exceeding 100 h, indicating exceptional stability under ambient conditions. Among the evaluated colorimetric methods, the DPD-based method was found to be suitable for quantifying ClOO•, showing a wide detection range and excellent linearity. This study represents the first report of a colorimetric quantification method for ClOO•. Our findings are expected to be useful for quantitatively discussing the biological efficacy of ClOO• and its reaction mechanisms.

## Linked entities

- **Chemicals:** chlorous acid (PubChem CID 24453), chloric acid (PubChem CID 19654), hydrogen peroxide (PubChem CID 784), ClOO• (PubChem CID 5460593), TMB (PubChem CID 41206)

## Full-text entities

- **Chemicals:** chloric acid (MESH:C022842), hydrogen peroxide (MESH:D006861), CAW (-), Na+ (MESH:D012964), DPPH (MESH:C004931), NaClO3 (MESH:C032706), chlorine (MESH:D002713), DPD (MESH:C036020)

## Full text

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## Figures

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## References

40 references — full list in the complete paper: https://tomesphere.com/paper/PMC12539726/full.md

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Source: https://tomesphere.com/paper/PMC12539726