The invention is concerned with the method of preparation of the Mydriasis ophthalmic preparation. It is the kind of ophthalmic preparation that in liquid form before dripping into eyes but turns to gelatin postural after. It contents Mydriasis agent as the active material and other supplementary materials, such as poloxamer, carbomer, Deacetylcephalomannine Gellan Gum, transition regulator and medicinal acceptable antiseptic, isoosmotic adjustment, PH regulator and injection usage water. It can use of optometry, pre-operation naydriasis, etc. The advantages of the invention are: highly biocompatibility, low pungency, and long lasting.