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High-sensitivity isopropanol Biomeasurement using enzyme cycling reactions.
Geng Zhang1, Kenta Ichikawa1, Kenta Iitani1
1Department of Biomedical Devices and Instrumentation, Laboratory for Biomaterials and Bioengineering, Institute of Integrated Research, Institute of Science Tokyo, 2-3-10 Kanda-Surugadai, Chiyoda-ku, Tokyo, 101-0062, Japan.
Talanta
|December 17, 2025
Summary
This study developed a sensitive enzyme cycling system to detect isopropanol (IPA) in biological fluids. This breakthrough enables non-invasive ketoacidosis monitoring using saliva analysis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biotechnology
Background:
- Elevated isopropanol (IPA) levels in biological fluids can indicate ketoacidosis.
- Current detection methods may lack the sensitivity required for early diagnosis.
- Non-invasive biomarkers are needed for convenient patient monitoring.
Purpose of the Study:
- To develop a highly sensitive detection system for isopropanol (IPA).
- To utilize enzyme cycling with secondary alcohol dehydrogenase (S-ADH) for enhanced detection.
- To establish a method for non-invasive ketoacidosis monitoring via saliva analysis.
Main Methods:
- Developed an enzyme cycling reaction system using S-ADH and a dual-coenzyme mechanism (thio-NAD+/NADH).
- Optimized reaction conditions including pH and coenzyme concentrations.
- Measured accumulating thio-NADH spectrophotometrically at 405 nm.
Main Results:
- Achieved a 6-fold increase in sensitivity compared to single-step detection.
- Expanded the detection range to 3.9 μM-1 mM, covering physiological IPA levels.
- Demonstrated successful detection within the physiological range of human saliva IPA concentrations (5.51-24.6 μM).
Conclusions:
- The optimized enzyme cycling system provides high sensitivity for IPA detection.
- This method facilitates non-invasive ketoacidosis monitoring through saliva analysis.
- Enzyme cycling biosensors show significant potential for clinical applications.
