As a highly active amphotropic compound in the ocean, hydrogen peroxide plays an important role on many marine biochemical reactions. Hydrogen peroxide in seawater is mainly photochemically produced in situ, and its concentration in seawater is generally decreasing from surface water to deep water, from estuaries to ocean, with the normal concentration range as 0 - 500 nM. Due to the characteristics of low abundance and short life, the detection of hydrogen peroxide in seawater requires a very low detection limit and high accuracy. The fluorescence analysis is one of the most commonly used quantitative detection methods for H2O2. In this study, we optimized the Scopoletin-HRP fluorescence analysis that used scopoletin as fluorescent reagent and horseradish peroxidase (HRP) as catalyst. We examined the effects of measurement interval, reagent pH, and reagent blank, on the detection of H2O2. For the first time, we applied three dimentional fluorescence technology to optimize the detection wavelengths of the Scopoletin-HRP method. The experimental data of this study showed that the best detection excitation wavelength is 385 nm, the best detection emission wavelength is 460 nm. This result has the potential to reduce the detection limit and to improve the accuracy of the method, showing a new direction for the further optimization research of this method. It also provides a certain reference for the subsequent studies that need to use the fluorescence analysis method to detect the hydrogen peroxide content in water bodies.
 

seawater,hydrogen peroxide,fluorography,scopoletin