Skip to main content
Article
Diagnosis of Glutaric Aciduria Type 1 by Measuring 3-hydroxyglutaric Acid in Dried Urine Spots by Liquid Chromatography Tandem Mass Spectrometry
Journal of Inherited Metabolic Disease
  • Osama Y. Al-Dirbashi, Children's Hospital of Eastern Ontario, Ottawa, ON
  • Stefan Kölker, University Children's Hospital, Heidelberg, Germany
  • Dione Ng, Children's Hospital of Eastern Ontario, Ottawa, ON
  • Lawrence Fisher, Children's Hospital of Eastern Ontario, Ottawa, ON
  • Tony Rupar, The University of Western Ontario
  • Nathalie Lepage, University of Ottawa
  • Mohamed S. Rashed, Pharmagene Labs, Giza, Egypt
  • Tomofumi Santa, University of Tokyo, Tokyo, Japan
  • Stephen I. Goodman, University of Colorado
  • Michael T. Geraghty, Children's Hospital of Eastern Ontario, Ottawa, ON
  • Johannes Zschocke, Innsbruck Medical University, Innsbruck, Austria
  • Ernst Christensen, National University Hospital Rigshospitalet, Copenhagen, Denmark
  • Georg F. Hoffmann, University Children's Hospital, Heidelberg, Germany
  • Pranesh Chakraborty, Children's Hospital of Eastern Ontario, Ottawa, ON
Document Type
Article
Publication Date
2-1-2011
URL with Digital Object Identifier
http://dx.doi.org/10.1007/s10545-010-9223-2
Disciplines
Abstract

Accumulation of glutaric acid (GA) and 3-hydroxyglutaric acid (3HGA) in body fluids is the biochemical hallmark of type 1 glutaric aciduria (GA1), a disorder characterized by acute striatal degeneration and a subsequent dystonia. To date, methods for quantification of 3HGA are mainly based on stable isotope dilution gas chromatography mass spectrometry (GC-MS) and require extensive sample preparation. Here we describe a simple liquid chromatography tandem MS (LC-MS/MS) method to quantify this important metabolite in dried urine spots (DUS). This method is based on derivatization with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole (DAABD-AE). Derivatization was adopted to improve the chromatographic and mass spectrometric properties of the studied analytes. Derivatization was performed directly on a 3.2-mm disc of DUS as a sample without extraction. Sample mixture was heated at 60°C for 45 min, and 5 μl of the reaction solution was analyzed by LC-MS/MS. Reference ranges obtained were in excellent agreement with the literature. The method was applied retrospectively for the analysis of DUS samples from established low- and high-excreter GA1 patients as well as controls (n = 100). Comparison of results obtained versus those obtained by GC-MS was satisfactory (n = 14). In populations with a high risk of GA1, this approach will be useful as a primary screening method for high- or low-excreter variants. In these populations, however, DUS analysis should not be implemented before completing a parallel comparative study with the standard screening method (i.e., molecular testing). In addition, follow-up DUS GA and 3HGA testing of babies with elevated dried blood spot C5DC acylcarnitines will be useful as a first-tier diagnostic test, thus reducing the number of cases requiring enzymatic and molecular analyses to establish or refute the diagnosis of GA1.

Citation Information
Osama Y. Al-Dirbashi, Stefan Kölker, Dione Ng, Lawrence Fisher, et al.. "Diagnosis of Glutaric Aciduria Type 1 by Measuring 3-hydroxyglutaric Acid in Dried Urine Spots by Liquid Chromatography Tandem Mass Spectrometry" Journal of Inherited Metabolic Disease Vol. 34 Iss. 1 (2011) p. 173 - 180
Available at: http://works.bepress.com/tony-rupar/2/