The invention belongs to the technical field of biochemical engineering and discloses a 1-carboxy alkyl-4-nitroimidazole preparation method and the application of the method. 6-bromohexanoic acid and thionyl chloride are mixed by a molar ratio of 1:1.5, and reflux reaction is carried out on the 6-bromohexanoic acid and the thionyl chloride, and compound methyl 6-bromohexanoate is prepared; 2-methyl-4- nitroimidazole and sodium hydride are dissolved in DMF by a molar ratio of 1:1.2, the compound 6-methyl 6-bromohexanoate with the same molar weight as the 2-methyl-4- nitroimidazole is added into the DMF to prepare 2-methyl-4-nitryl-1-(6-carboxyl hexyl) imidazate; the imidazate is hydrolyzed in a sodium hydroxide aqueous solution to prepare 1-carboxy alkyl-4-nitroimidazole. According to the method, the synthesis steps are simple and effective, the product is high in purity after being identified, and the requirement for immune research on nitroimidazole drugs of China can be met.