Enantioseparation of d,l-2-hydroxyglutaric acid by capillary electrophoresis with tandem mass spectrometry-Fast and efficient tool for d- and l-2-hydroxyglutaracidurias diagnosis

J Chromatogr A. 2016 Oct 7:1467:383-390. doi: 10.1016/j.chroma.2016.05.095. Epub 2016 May 27.

Abstract

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.

MeSH terms

  • Adult
  • Amino Acid Metabolism, Inborn Errors / diagnosis*
  • Amino Acid Metabolism, Inborn Errors / urine
  • Child
  • Creatinine / urine
  • Electrophoresis, Capillary*
  • Glutarates / isolation & purification*
  • Glutarates / urine
  • Humans
  • Mass Spectrometry*
  • Reproducibility of Results
  • Stereoisomerism
  • Tandem Mass Spectrometry*

Substances

  • Glutarates
  • alpha-hydroxyglutarate
  • Creatinine