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.
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.
Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacký University, in Olomouc, 17. listopadu 12, Olomouc CZ-77146, Czech Republic.
Martin
M
Švidrnoch
Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacký University, in Olomouc, 17. listopadu 12, Olomouc CZ-77146, Czech Republic.
Adam
A
Přibylka
Laboratory for Inherited Metabolic Disorders, Department of Clinical Biochemistry, University Hospital, I. P. Pavlova 6, CZ-77520 Olomouc, Czech Republic; Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University in Olomouc, Hněvotínská 5, CZ-77900 Olomouc, Czech Republic.
Vojtěch
V
Bekárek
Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacký University, in Olomouc, 17. listopadu 12, Olomouc CZ-77146, Czech Republic.
Juraj
J
Ševčík
Child Clinic, Faculty of Medicine, Palacký University in Olomouc, Hněvotínská 3, Olomouc, Czech Republic.
Vratislav
V
Smolka
Regional Centre of Advanced Technologies and Materials, Department of Analytical Chemistry, Faculty of Science, Palacký University, in Olomouc, 17. listopadu 12, Olomouc CZ-77146, Czech Republic. Electronic address: vitezslav.maier@upol.cz.
Vítězslav
V
Maier
D000328:Adult; D000592:Amino Acid Metabolism, Inborn Errors / Q000175:diagnosis* / Q000652:urine; D002648:Child; D003404:Creatinine / Q000652:urine; D019075:Electrophoresis, Capillary; D005977:Glutarates / Q000302:isolation & purification* / Q000652:urine; D006801:Humans; D013058:Mass Spectrometry; D015203:Reproducibility of Results; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry
D005977:Glutarates; C019417:alpha-hydroxyglutarate; D003404:Creatinine
2-Hydroxyglutaric aciduria; Capillary electrophoresis; Chiral separation; Mass spectrometry; Vancomycin