A modified analytical procedure involving gas-liquid chromatography was used to investigate the relative amounts of 2,4-D [(2,4-dichlorophenoxy)acetic acid] and two of its amino acid conjugate metabolites, 2,4-D-Asp [(2,4-dichlorophenoxy)acetylaspartic acid] and 2,4-D-Glu [(2,4-dichlorophenoxy)acetylglutamic acid], in soybean [Glycine max (L.) Merr. var. Amsoy] plants, soybean callus tissue and red oak (Quercus rubra L.) plants at 2, 4, and 10 days after application of either 2,4-D, its ester, or its salts. 2,4-D-Asp and 2,4-D-Glu were found as major metabolites in all soybean tissues analyzed and their amounts varied depending upon the days following treatment, the formulation of treatment, and the plant substrate (0.03 to 57 ppm for 2,4-D-Asp and 0.05 to 75 ppm for 2,4-D-Glu). The amount of amino acid conjugates peaked at 2 days after treatment of callus tissue and 4 days after treatment of soybean plants. Much higher amounts of these amino acid conjugates were obtained from soybean plants treated with the propylene glycol butyl ether ester of 2,4-D than plants treated with amine salts of 2,4-D. No amino acid conjugates were observed with red oak plants.