A Quantitative Determination of Singlet Oxygen with Horseradish Peroxidase

A singlet oxygen determination method based on the formation of dimethyl-(2,5)-2-methoxy-5-hydroperoxydihydrofuran (DMFO2) followed by acid hydrolysis with 0.1 N H2SO4is described. Hydrogen peroxide is formed thereby as a product of hydrolysis which is then determined by the horseradish peroxidase-catalyzed formation of the homovanillic acid dimer. Since DMFO2is a crystalline compound with sufficient stability it can be used as a primary standard for the determination. The peroxidase-catalyzed hydrogen peroxide determination allows detection of singlet oxygen concentrations as low as 10−7M.