title: Development of a new tandem mass spectrometry method for urine and amniotic fluid screening of oligosaccharidoses.
abstract: BACKGROUND
  The first step in the diagnosis of oligosaccharidoses is to evidence abnormal oligosaccharides excreted in urine, usually performed by the poorly sensitive but efficient thin layer chromatography (TLC) method. Developing a tandem mass spectrometry (MS/MS) technique could be of great interest to replace TLC.
  
  
  METHODS
  Abnormal underivatized oligosaccharides have been recently studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, allowing the unambiguous identification of oligosaccharidoses. Based on this previous work, we developed an advantageous and efficient liquid chromatography (LC)/MS/MS method using a more common triple quadrupole tandem mass spectrometer for oligosaccharides analysis.
  
  
  RESULTS
  Oligosaccharidoses (n = 97) and control (n = 240) urine samples were analysed. A specific pattern was obtained for each oligosaccharidosis using this method. In urine, it allows not only the identification of all the oligosaccharidoses previously identified by TLC (fucosidosis, alphamannosidosis, aspartylglucosaminuria, GM1 gangliosidosis, sialidosis, galactosialidosis and Schindler disease), but also extends the field of diagnosis to mucolipidosis type II, Sandhoff disease, and β-mannosidosis. The same technique was applied to 16 amniotic fluid supernatants from oligosaccharidosis-affected foetuses (n = 16) compared with 37 unaffected. All the affected foetuses could be clearly identified: sialidosis (n = 3), galactosialidosis (n = 4), aspartylglucosaminuria (n = 1), mucolipidosis type II (n = 4) or GM1 gangliosidosis (n = 4). This technique can be applied to early prenatal diagnosis as well as to the oligosaccharidosis screening in the case of non-immune hydrops fetalis.
  
  
  CONCLUSIONS
  The method is quick and easy to run, with an LC analysis time of 13 min per sample. The quantitative validation could not be obtained in the absence of a specific standard and of a labelled internal standard for each compound. Even if this LC/MS/MS method is only qualitative, it is very specific and much more sensitive than TLC. It allows the urinary screening of oligosaccharidoses, even mild or late-onset forms, and the screening of antenatal forms in amniotic fluid. Copyright © 2017 John Wiley & Sons, Ltd.
authors.affiliation: Service de Biochimie et Biologie Moléculaire Grand Est, UM Pathologies Métaboliques, Erythrocytaires et Dépistage Périnatal, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, France.
authors.forename: Monique
authors.identifier: http://orcid.org/0000-0002-2341-6514
authors.initials: M
authors.lastname: Piraud
authors.affiliation: Service de Biochimie et Biologie Moléculaire Grand Est, UM Pathologies Métaboliques, Erythrocytaires et Dépistage Périnatal, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Pettazzoni
authors.affiliation: Service de Biochimie et Biologie Moléculaire Grand Est, UM Pathologies Métaboliques, Erythrocytaires et Dépistage Périnatal, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, France.|Laboratoire de Biochimie Médicale, Centre Hospitalo-Universitaire François Mitterrand, Dijon, France.
authors.forename: Louise
authors.initials: L
authors.lastname: Menegaut
authors.affiliation: Laboratoire de Biochimie Métabolomique et Protéomique, Hôpital Universitaire Necker-Enfants Malades, Assistance Publique-Hôpitaux de Paris, France ; Unité INSERM 1151, Université Paris Descartes, Paris, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Caillaud
authors.affiliation: Département de Neurologie, Hôpital Pitié-Salpêtriére, Assistance Publique des Hôpitaux de Paris, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Nadjar
authors.affiliation: Service de Biochimie et Biologie Moléculaire Grand Est, UM Pathologies Métaboliques, Erythrocytaires et Dépistage Périnatal, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, France.|Unité INSERM U1060, Laboratoire CarMeN, Université Lyon-1, Lyon, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Vianey-Saban
authors.affiliation: Service de Biochimie et Biologie Moléculaire Grand Est, UM Pathologies Métaboliques, Erythrocytaires et Dépistage Périnatal, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, France.|UMR 5305 CNRS/UCBL, Lyon, France.
authors.forename: Roseline
authors.initials: R
authors.lastname: Froissart
mesh_terms: D000653:Amniotic Fluid / Q000737:chemistry*; D005260:Female; D006801:Humans; D016014:Linear Models; D016464:Lysosomal Storage Diseases / Q000175:diagnosis*; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000652:urine; D011296:Prenatal Diagnosis / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009844:Oligosaccharides