Abstract: OBJECTIVE To optimize the preparation process of Fule suppository and establish its assay method. METHODS Orthogonal design, the appearance, weight difference, disintegration, dissolution, content as index, the effects of different concentration of alcohol, polyethylene glycol ratio, temperature, stirring speed, influence of suppository preparation the process time and casting temperature and stirring. The content of berberine hydrochloride in Shuanghuang Fule suppository was determined by HPLC. RESULTS The best preparation condition was PEG-400:PGE-6000:S-40=6:3:1, temperature of 50℃, stirring time of 40 min, stirring speed of 150 r·min^-1, injection temperature of 60℃. The berberine hydrochloride content of Shuanghuang Fule suppository were 2.10, 2.14, 2.15 mg. CONCLUSION The optimal proportion of the prescription is reasonable, the preparation process is stable and easy to operate, and the content determination method is reliable.