HPLC Separation of Isomers of Aminobenzoic Acid

Isomers of organic compounds usually have very similar properties. Separation of such isomers by HPLC requires intensive screening of various columns and mobile phase conditions. In case of hydrophilic compounds, this task becomes even more challenging due to the lack of or limited retention on traditional reversed-phase (C18 or C8) HPLC columns. Three isomers of aminobenzoic acid (2-aminobenzoic acid, 3-aminobenzoic acid and 4-aminobenzoic acid) were successfully separated on a Primesep 100 reversed-phase/cation-exchange column. Baseline separation within 10 minutes was achieved and produced a perfect peak shapes for all three analytes. The method can be used a generic approach for separation of ionic hydrophobic and ionic basic isomers by mixed-mode chromatography.