The invention relates to a sterilization disinfectant used for a clinical laboratory and a preparation method of the sterilization disinfectant. Raw materials for preparing effective constituents of the disinfectant comprises the bark of ash, sculellaria barbata, fisheye grass, saururus chinensis, prismatomeria tetrandra, large-leaved gentian, oriental wormwood, spina gleditsiae, cudrania cochinchinensis kudo et masam, eupatorium, fructus ulmi, Japan clover herb, witloof, cottonrose hibiscus, folium sennae and wikstroemia indica. Compatibility of medicinal materials selected by the disinfectant is appropriate, and the disinfectant is prepared by adopting modern advanced medicine pharmaceutical technology, strong in sterilization ability, fast in action speed and strong in stability, and the like, does not have stimulation on human skin, is soluble in water, does not have residue after use, is safe and doe not have toxic and side effects. The sterilization disinfectant can be widely applied to disinfection of staff skin, instruments, and the like of the clinical laboratory. By utilizing the sterilization disinfectant, the work efficiency of clinical laboratory staff is improved, the accuracy of inspection result is ensured, and the sterilization disinfectant is an ideal disinfectant worthy to be popularized.