The safety of tooth bleaching, which is based upon hydrogen peroxide (HP) as the active agent, has been questioned. Our aim was to investigate the effects of 30% HP on human tooth enamel. The specimens were divided randomly into three groups and treated with distilled water, HCl, and HP, respectively. Raman scattering and laser-induced fluorescence of enamel were determined before and after treatment. Microhardness testing and scanning electron microscopy were also used. The results of Raman scattering showed that the Raman relative intensity of enamel changed significantly after HP and HCl treatment. These findings were consistent with the results of microhardness testing and morphological observations. In addition, a small band at 876 cm-1 due to O–O stretching of HP became pronounced during HP treatment, which provided direct evidence that HP has the ability to penetrate enamel. Meanwhile, the results of laser-induced fluorescence revealed that HP caused the greatest fluorescence reduction. This suggested that the organic matter in enamel might be greatly affected by HP, which was also supported by the results of microhardness. It can be concluded, therefore, that the 30% HP may have adverse effects on the mineral and the organic matter of human tooth enamel.