The invention relates to the fields of daily use chemicals and pharmaceutical industry, in particular to carbamide-containing acetylated chitosan quaternary ammonium salt, and a preparation method and application thereof. The preparation method of the carbamide-containing acetylated chitosan quaternary ammonium salt is characterized in that firstly, chitosan reacts with iodomethane and sodium iodide under the alkaline condition for a period of time, and then reacts with chloroacetyl chloride to obtain chloracetylated chitosan quaternary ammonium salt; then, various types of phenylamine and triphosgene react to prepare isocyanate; the isocyanate and 4-aminopyridine react to prepare carbamide; finally, the chloracetylated chitosan quaternary ammonium salt and the prepared carbamide react; through purification, the carbamide-containing acetylated chitosan quaternary ammonium salt is obtained, wherein the molar weight of the chloracetyl chloride is 1 to 2 times of that of the chitosan; the molar weight of the various types of carbamide is 2 to 3 times of that of the chloracetylated chitosan quaternary ammonium salt. The reaction is simple, convenient and efficient; the popularization is easy; the required equipment and raw materials can be easily obtained. The research shows that the derivative has good water solubility and good bacteriostatic activity; the biological activity of the chitosan is enhanced; the application range of the chitosan is enlarged; the chitosan quaternary ammonium salt can be widely applied to the fields of daily use chemicals and medicine.
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