The invention provides a pharmaceutical composition with effects of promoting blood circulation to remove blood stasis and a preparation method and application thereof, and belongs to the field of medicine. The composition comprises a basic material and a traditional Chinese medicine mixture; the basic material is prepared from acetin and steel materials; the traditional Chinese medicine mixture is prepared from radix aconiti, radix aconite agrestis, mastic, myrrh, pseudo-ginseng, ligusticum wallichii, semen plantaginis, resina draconis, sappanwood, radix angelicae pubescentis, radix curcumaelongae, rhizoma corydalis, rhizoma acori graminei and garden balsam stems. The preparation method comprises the steps that the traditional Chinese medicine mixture is soaked with basic liquid and placed for 0.25-24 days away from light, and filtering is performed, wherein the basic liquid is obtained after the acetin and the steel materials are mixed, sealed, placed, cellared for 6-8 h and filtered. The pharmaceutical composition is good in transdermal absorption effect, has the good curative effect on treating cervical spondylosis, lumbar disc herniation and scapulohumeral periarthritis, andhas no side effect.