A study of the Bratton and Marshall hydrolysis procedure utilizing high performance liquid chromatography

Most analytical methods for conjugated aromatic amines require hydrolysis to free the amine group. The rate of conversion of N-acetyl-p-aminohippuric acid to its deacetylated and deglycinated derivatives is directly related to the hydrolysis reaction time and the concentration of hydrochloric acid used in the reaction. Variations in methodology involving either of these hydrolysis parameters produced artifacts which were not detectable by the standard colorimetric procedures. High performance liquid chromatography offers an improved tool for detecting and quantitating the changes produced during hydrolysis.