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.