The invention discloses an extraction and preparation method of streptomycin sulfate. The extraction and preparation method comprises the steps of acidizing, filtering, adsorbing, pre-washing, eluting, decompression concentrating, salt forming, decoloring, concentrating and spray-drying, thus obtaining a finished product. The extraction and preparation method of the streptomycin sulfate has the beneficial effects that the technical problems in the prior art that the extraction and preparation route is long, the production efficiency is low, the investment cost is high, the yield is low, the product color is poor, and the qualification rate is low are solved and has the advantages that the extraction and preparation process is short, the procedures are simple, the production efficiency is high, the investment cost is low, the consumption of active carbon is small, the yield is high, the product color is good, and the qualification rate is high.