The invention discloses an antibacterial formula containing vegetable medicinal oil as well as a preparation method and application of the antibacterial formula. The antibacterial formula containing the plant medicinal oil is prepared from the following raw materials in parts by mass: 10-15 parts of cinnamomum camphora leaf volatile oil, 1-10 parts of mint volatile oil, 5-15 parts of olibanum oil,5-15 parts of cinnamon bark volatile oil, 1-10 parts of gallnut oil, 10-20 parts of michelia alba volatile oil, 1-10 parts of lemongrass volatile oil and 1-10 parts of lemon volatile oil. 1-10 partsof sandalwood volatile oil, 1-10 parts of Mentha longifolia volatile oil, 1-10 parts of Roman chamomile volatile oil, 1-10 parts of radix angelicae volatile oil, 1-10 parts of clary sage volatile oil,1-10 parts of gardenia volatile oil, 5-10 parts of cananga odorata volatile oil, 1-10 parts of eucommia seed oil, 5-15 parts of eucalyptus leaf volatile oil, 1-10 parts of honeysuckle volatile oil, 5-10 parts of purslane volatile oil and 2-5 parts of dandelion volatile oil. The beneficial effects are that the antibacterial formula is pleasant in fragrance, and has the effects of inhibiting bacteria, purifying air and regulating emotion, and is small in toxic and side effects and high in safety.