Enantiomeric Separation of 2-Hydroxyglutarate Using Chiral Mobile Phase Additives

Abstract

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.

An on-line heart-cutting two-dimensional liquid chromatography method for intracellular 2-hydroxyglutarate enantiomers

“…A chiral mobile phase method for 2-HG enantiomers was recently developed using copper( ii ) acetate and N , N -dimethyl- l -phenylalanine as chiral mobile phase additives. 20 This method requires no derivatization, but suffers from a bad peak shape and low sensitivity.…”