Diagnosis of glutaric aciduria type 1 by measuring 3-hydroxyglutaric acid in dried urine spots by liquid chromatography tandem mass spectrometry
Osama Y. Al-Dirbashi
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Department of Pediatrics, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Search for more papers by this authorStefan Kölker
Department of General Pediatrics, University Children's Hospital, Im Neuenheimer Feld 430, Heidelberg, 69120 Germany
Search for more papers by this authorDione Ng
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Search for more papers by this authorLawrence Fisher
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Search for more papers by this authorTony Rupar
Biochemical Genetics Laboratory, London Health Sciences Centre, London, Ontario, Canada
Search for more papers by this authorNathalie Lepage
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Search for more papers by this authorTomofumi Santa
Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan
Search for more papers by this authorStephen I. Goodman
University of Colorado Health Sciences Center, Aurora, Colorado, USA
Search for more papers by this authorMichael T. Geraghty
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Department of Pediatrics, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Search for more papers by this authorJohannes Zschocke
Division of Human Genetics, Innsbruck Medical University, Innsbruck, Austria
Search for more papers by this authorErnst Christensen
National University Hospital Rigshospitalet, Blegdamsvej 9, Copenhagen, 2100 Denmark
Search for more papers by this authorGeorg F. Hoffmann
Department of General Pediatrics, University Children's Hospital, Im Neuenheimer Feld 430, Heidelberg, 69120 Germany
Search for more papers by this authorCorresponding Author
Pranesh Chakraborty
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Department of Pediatrics, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Telephone: +1-613-7377600, [email protected]
Search for more papers by this authorOsama Y. Al-Dirbashi
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Department of Pediatrics, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Search for more papers by this authorStefan Kölker
Department of General Pediatrics, University Children's Hospital, Im Neuenheimer Feld 430, Heidelberg, 69120 Germany
Search for more papers by this authorDione Ng
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Search for more papers by this authorLawrence Fisher
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Search for more papers by this authorTony Rupar
Biochemical Genetics Laboratory, London Health Sciences Centre, London, Ontario, Canada
Search for more papers by this authorNathalie Lepage
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Search for more papers by this authorTomofumi Santa
Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan
Search for more papers by this authorStephen I. Goodman
University of Colorado Health Sciences Center, Aurora, Colorado, USA
Search for more papers by this authorMichael T. Geraghty
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Department of Pediatrics, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Search for more papers by this authorJohannes Zschocke
Division of Human Genetics, Innsbruck Medical University, Innsbruck, Austria
Search for more papers by this authorErnst Christensen
National University Hospital Rigshospitalet, Blegdamsvej 9, Copenhagen, 2100 Denmark
Search for more papers by this authorGeorg F. Hoffmann
Department of General Pediatrics, University Children's Hospital, Im Neuenheimer Feld 430, Heidelberg, 69120 Germany
Search for more papers by this authorCorresponding Author
Pranesh Chakraborty
Newborn Screening Ontario, Children's Hospital of Eastern Ontario, 401 Smyth Road, Ottawa, ON, K1H 8L1 Canada
Department of Pediatrics, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Rd, Ottawa, Ontario, Canada
Telephone: +1-613-7377600, [email protected]
Search for more papers by this authorCommunicated by: Verena Peters
Competing interest: None declared.
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.
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