Chiral liquid chromatography tandem mass spectrometry in the determination of the configuration of 2-hydroxyglutaric acid in urine.

D-2-Hydroxyglutaric aciduria and L-2-hydroxyglutaric aciduria are two distinct inherited metabolic diseases. The accurate diagnosis of the exact disorder relies on the determination of the configuration of the enantiomers, either D-2-hydroxyglutaric acid or L-2-hydroxyglutaric acid excreted in excess in urine of patients. The enantiomeric chiral separation of 2-hydroxyglutaric acid was achieved using a ristocetin A glycopeptide antibiotic silica gel bonded column. The chiral column was interfaced with a tandem mass spectrometer for the purpose of specifically detecting the eluting 2-hydroxyglutaric acid. Tandem mass spectrometry was employed using an electrospray ion source in the negative ion mode. Three parent-to-daughter transitions under collision-induced dissociation conditions were used to detect only 2-hydroxyglutaric acid. The two forms of the compound were satisfactorily separated with almost baseline resolution at 4.95 and 5.5 min. Three known patients with 2-hydroxyglutaric aciduria were identified to have L-2-hydroxyglutaric aciduria. The method is simple, selective, rapid, and free from interference. Copyright 2000 John Wiley & Sons, Ltd.