Aim To pick out the best type of penetration enhancers through the test in vitro using excised sheep nasal mucosa.Methods Diffusion cells were adopted as apparatus for in vitro nasal mucosa.The concentration of dihydroetorphine hydrochloride was measured by HPLC and the cumulative penetration amount was calculated.Results The cumulative penetration amount of dihydroetorphine hydrochloride was linearly related to time.The penetration rate was significantly increased when penetration enhancers were used.Of these penetration enhancers,DM-β-CD was the best,and HP-β-CD and propylene glycol were the second best,followed by β-CD.Conclusion Penetration enhancers could enhance the penetration rate of dihydroetorphine hydrochloride,especially when DM-β-CD was added.