Abstract: An experimental method for determination of the content of gliclazide in Dameikang by ¹H-NMR was established. With dimethyl terephthalate as internal standard substance and deuterated chloroform as solvent, the experimental parameters were established through gliclazide standard substance and dimethyl terephthalate under a Bruker AVANCE II 400 nuclear magnetic resonance spectrometer. The standard curve was drawn and the precision was studied. The content of gliclazide in Dameikang was determined by this experimental parameter. It turned out the experimental results were consistent with the contents in the manual and results by HPLC. The method established in this paper is accurate and rapid, which provides a new method for the quality control and reference material calibration of this variety.