In the present paper, DMSO/H2O mixture with the ratio of volume 1∶1 was measured in the cooling process by Raman spectroscopy, and the Raman assignments was made to the DMSO molecular and water molecular. The results showed that the behavior between intra-molecular hydrogen bonds and inter-molecular hydrogen bonds of DMSO and water leads to the change in the Raman spectra of the SO stretching vibration of DMSO and the O—H stretching vibration of water. Further analysis showed that the hydrogen bond between DMSO and water was enhanced in the course of temperature decreasing process (27 to -30 ℃), and the intramolecular hydrogen bonds between water and water replaced the intermolecular hydrogen bonds of DMSO and water in the course of temperature decreasing process (-30 to -60 ℃). The research provides experimental basis for hydrogen bonding theory in aqueous solution.