The invention relates to an operating room disinfectant and a preparation method, and belongs to the field of traditional Chinese medicines. The disinfectant is prepared from the effective ingredients of leaves of camphor trees, herba portulacae, leaves and twigs of nakedflower beautyberry, uncinata spikemoss herb, fructus forsythiae, herb of Japanese youngia, horseweed herb, purple flower holly leaves, birch bark and emblic leafflower bark. The operating room disinfectant is high in pertinence and is prepared from traditional Chinese medicine plants of leaves of camphor trees, herba portulacae, leaves and twigs of nakedflower beautyberry, uncinata spikemoss herb, fructus forsythiae, herb of Japanese youngia, horseweed herb, purple flower holly leaves, birch bark and emblic leafflower bark, disinfection and decontamination are combined, and the disinfectant has a high bactericidal capacity for multiple bacteria such as staphylococcus aureus, candida albicans, pseudomonas aeruginosa, bacillus subtilis and escherichia coli and is wide in antibacterial spectrum, long-acting in disinfection and high in bacteriostatic property.