Indirect enantiomeric separation of 2-arylpropionic acids and structurally related compounds by reversed phase HPLC

Abstract

A reversed-phase high-performance liquid chromatographic method, using an organic modifier-phosphate buffered mobile phase, for the determination of the enantiomeric composition of 2-arylpropionic acids and other structurally related compounds in microbial media is described. The method is based on the resolution of diastereoisomeric amides formed from the reaction of the arylpropionic acid with either (-)-(S)-alpha-methylbenzylamine or (-)-(S)-1-(naphthen-1-yl)ethylamine in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide HCl and 1-hydroxybenzotriazole and incorporating an internal standard. The addition of sodium pentanesulphonate to the mobile phase as an ion-pairing agent was necessary to remove unreacted amine to avoid rapid column deterioration. The method provides and efficient, rapid and reproducible means of monitoring the microbial chiral inversion of 2-arylpropionic acids and other structurally related molecules.