The N2O formation pathway and effect of H2O on N2O formation during the NOx storage and reduction (NSR) process using CO as a reductant were investigated over a Pt-BaO/Al2O3 catalyst. The NSR activity measurements and transient in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) experiments were performed to evaluate N2O evolution and elucidate the N2O formation mechanism. N2O is formed in the lean, rich, and delay2 phases. In the lean phase, N2O formation is related to the reactions between surface isocyanate and gaseous NO/O2 and NO is more responsible for N2O formation than O2. Moreover, N2O production decreases with H2O because of the hydrolysis of isocyanate species. In the rich phase, the amount of N2O formation also decreases in the presence of H2O at a higher temperature because of the high reduction ability of H2 generated from the water-gas shift (WGS) reaction. During the delay2 phase, N2O is mainly formed by nitrite species reacting with Pt(0)-CO. Furthermore, the presence of H2O decreases the stability of nitrites and results in more N2O production at a low temperature.