The present paper introduces a new method using spectrofluorimetric analysis to determine the concentration of hydrogen peroxide in rainwater. In this method, an oxidation reaction is conducted between o-phenylenediamine (OPDA) and hydrogen peroxide in the buffer medium of NaAc-HAc at pH 4. 48 to form a new product 2,3-diaminophenazine (DAPN). Then the fluorescence intensity of DAPN is measured and 426 and 554 nm are chosen as the excitation and emission wavelengths. Therefore, with the foreknown concentration of input hydrogen peroxide, a series of fluorescence intensities of DAPN are acquired according to a series of different concentration of hydrogen peroxide as input, greatly improving the selectivity and sensibility of the system. A relationship between the input concentration of hydrogen peroxide and the fluorescence intensity of DAPN is then obtained using a linear regression. Results show that fluorescence intensity of DAPN is in proportion to the increase in the concentration of hydrogen peroxide in the range of 9.0 x 10(-7) -3.56 x 10(-5) mol x L(-1) almost linearly. The linear equation is F = 1.15c (micromol x L(-1))+398.6 (r = 0.999 1) and the detection limit is 2.7 x10(-7) mol x L(-1) (n = 11). The relative standard deviation of 11 parallel measurements with the concentration of H2O2 at 7.5 x 10(-6) and 3.0 x 10(-5) mol x L(-1), is 2.2 and 1.0%, respectively. Results from DPD method was used to verify this method. The interference of foreign iron was studied. Compared to the traditional methods, this binary system has a simplified operation and high sensitivity. The proposed method has been successfully applied to determine the concentration of hydrogen peroxide in rainwater.