Enantiomeric Separation of 2-Hydroxyglutarate Using Chiral Mobile Phase Additives

2-Hydroxyglutarate (2-HG) was enantiomerically separated using copper(II) acetate and N,N-dimethyl-L-phenylalanine as chiral additives. These compounds formed diastereomeric complexes with 2-HG, which were successfully separated on an achiral (ODS) column. Several parameters, such as additive concentration, the type of organic modifier, and column temperature, were optimized. Using the optimal mobile phase (1 mM copper(II) acetate and 2 mM N,N-dimethyl-L-phenylalanine in a 10% aqueous methanol solution), 2-HG enantiomers were successfully separated in 15 min with a resolution of 1.93.