Objective To optimize the formulation of Zushima Gel Paste by orthogonal test and study its trans-dermal absorption behavior in vitro.Methods The initial viscosity, paste strength, and release degree were used as indexes, and the dosage of blank matrix excipients was optimized by L_(18)(3∼7) orthogonal test to determine the optimal formulation combination and carry out validation test.The in vitro trans-dermal permeation of daphnetin, daphnetin-8-O-β-D-glucoside, and daphnetin in the optimal formulation was simultaneously determined by HPLC method using a modified Franz diffusion cell.Results The optimum formulation of Zushima Gel Paste was carbomer 980, polyvinyl K30, micronized silica gel, acacacia, polyvinyl K90, gelatin, and polyethylene glycol 400 with proportion of 8:3:4:12:8:10:8.The in vitro transdermal behavior conformed to the Zero-order kinetic process.Conclusion The gel paste prepared with the optimized adhesive formula has good adhesion, ointment strength, and drug release performance, and has good use comfort.It can be used as the basic preparation formula of Zushima Gel Paste, and can provide tech. reference for the development of new gel ointment.