OBJECTIVE To prepare itraconazole vaginal suppository and establish its quality control method. METHODS Gelatin，glycerin，polyethylene glycol，etc. were used as base materials and itraconazole as active ingredient to prepare the suppository. The content of suppository was determined with RP-HPLC，and the in vitro release was observed by improved suppository release determination method. RESULTS A good linearity was obtained from 10 to 100 μg·mL-1，r=0.999 9. The average recovery was 98.45%，and RSD was 1.37%. At 37 ℃，all suppositories melt within 1 h.The release parameter via Riger-Peppas equation simulation，n，was larger than 0.89，indicating that the drug release mechanism was erosion. CONCLUSION The suppository preparation method is easy. The determination is accurate. The in vitro release is accurately determined by improved suppository release determination method.