The purpose of this study was to investigate the chemical characteristics of dentin after Er:YAG laser irradiation using various output energies with or without water irrigation. Analysis was carried out by means of attenuated total reflectance Fourier transform infrared spectroscopy (FT-IR/ATR) and X-ray diffraction (XRD). Furthermore, the relative infrared peak intensities of dentin specimens were compared statistically. Results showed that Er:YAG laser with an output energy of 100 mJ/pulse with water irrigation did not cause any detectable change in dentin. However, a higher energy output or the absence of water irrigation affected the organic portion of dentin. With XRD, no obvious phase changes were observed between the XRD pattern of the control (non-irradiated) dentin powder and those after Er:YAG irradiation - regardless of Er:YAG laser output energy or dehydration condition. It was suggested that the intrinsic water content of dentin - together with extrinsic water irrigation - were important factors to achieving the desired outcome of dentin ablation by Er:YAG laser.