To provide a production method of 4-amino antipyrine capable of achieving good measurement of a biogenic substance keeping a designed level of sensibility when used for a biogenic substance measuring agent in an enzyme-peroxidase-coloring agent system.SOLUTION: A production method of purified 4-amino antipyrine that is used for a biogenic substance measuring agent keeping a designed level of sensibility in an enzyme-peroxidase-coloring agent system includes the following steps: (1) a step for preparing a solution dissolving crude 4-amino antipyrine in a solvent; and (2) a step for removing 4-hydroxy antipyrine by separating it from the solution making use of its affinity to a resin having a ligand.SELECTED DRAWING: None
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