The invention mainly provides a sanitizer formulation and a preparation method thereof. The sanitizer formulation comprises 6-12 parts of honeysuckle, 8-12 parts of fructus forsythiae, 6-12 parts of onion, 6-10 parts of houttuynia cordata, 8-12 parts of dandelion, 6-10 parts of radix bupleuri, 5-7 parts of hickory oil, 5-8 parts of radix isatidis, 15-22 parts of activated carbon, 6-8 parts of ginger, 5-9 parts of folium artemisiae argyi, 5-8 parts of eucalyptus leaf, 5-8 parts of radix sophorae flavescentis, 5-8 parts of glycerin, 6-10 parts of aloe, 10-15 parts of hydrogen peroxide, 3-5 partsof potassium permanganate, 8-15 parts of anhydrous alcohol, 5-15 parts of dimethyl formamide and 30-150 parts of distilled water. Honeysuckle, fructus forsythia, onion, houttuynia cordata, dandelion,radix bupleuri, radix isatidis, ginger, folium artemisiae argyi, the eucalyptus leaf, radix sophorae flavescentis and aloe which have sterilization and disinfection effect and are low in cost, easy to obtain and obvious in disinfection and sterilization effect are used as main raw materials, potassium permanganate, hydrogen peroxide and the hickory oil are used as auxiliary materials, and glycerin and dimethyl formamide are compounded, so that the prepared sanitizer formulation is higher in sterilization speed, stronger in sterilization effect, lower in sterilization concentration, nontoxic and pollution-free.