A novel capillary electrophoresis-tandem mass spectrometry method for the enantioseparation and identification of 2-hydroxyglutaric acid enantiomers without derivatization for clinical purposes was described. Vancomycin chloride was used as an efficient chiral selector for the discrimination of 2-hydroxyglutaric acid enantiomers by capillary electrophoresis employed complete capillary filling method. The obtained resolution was 2.05. Hyphenation of CE with tandem mass spectrometry allows a reliable identification of separated enantiomers as well as their quantification. The method was validated and applied for the separation, identification and determination of 2-hydroxyglutaric enantiomers in urine samples obtained from healthy patients and two urine samples obtained from child patients suffering from high urine excretion of 2-hydroxyglutaric acid. Abnormal excretion of d-hydroxyglutaric acid was found in both child urine samples (104.5±2.1 and 2200.0±12.6mmol/mol of creatinine, respectively). The limits of detection for d- and l-hydroxyglutaric acid were 31 and 38nmol/L, respectively.
Keywords: 2-Hydroxyglutaric aciduria; Capillary electrophoresis; Chiral separation; Mass spectrometry; Vancomycin.
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