This study was carried out to evaluate the effects of Chinese herbal transdermal ointment on mastitis in the dairy cow and to select the best efficient formula for treating the disorder. With chlorogenic acid as a marker, using high performance liquid chromatography (HPLC), the contents of chlorogenic acid in permeate liquid were determined. Different skin permeation enhancing effects of Chinese herbal medicinal formulations and various herbal transdermal promoters were evaluated in the form of ointment. The results indicated that ultrafined powder (UFP) of Chinese herbal medicine (CHM) directly adding to medication and 4.5% borneol plus 4.5% Azone as skin permeation enhancers showed the best skin-penetrating effect.