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Biological reduction of 3,5-dinitro-o-toluamide results in the formation of amino-nitro-otoluamide. The isomer isolated from tissue homogenates was identified by chemical and physical tests as the 3-amino-5-nitro-o-toluamide. The isomer which is generally obtained by the chemical reduction of 3,5-dinitro-o-toluamide is the 5-amino-3-nitro-otoluamide.

Develop the column by washing with four to five 250-ml. portions of TV hydrochloric acid. The dye derived from 0aminoacetophenone separates almost immediately, moving ahead of the other dyes, and washes completely through the column. The PCA dye is adsorbed in the lower half of the column and the DCA dye forms a completely separate zone at the top half.