Tandem Mass Spectrometry Quantitation of Lyso-Gb3 and Six Related Analogs in Plasma for Fabry Disease Patients

Curr Protoc Hum Genet. 2016 Jul 1:90:17.23.1-17.23.9. doi: 10.1002/cphg.4.

Abstract

Fabry disease is an X-linked lysosomal storage disorder, caused by a deficit in α-galactosidase A enzyme activity, leading to the storage of sphingolipids such as globotriaosylsphingosine (lyso-Gb3 ), globotriaosylceramide (Gb3 ), and galabiosylceramide (Ga2 ) in organs, tissues and biological fluids. A recent metabolomic study performed in plasma revealed lyso-Gb3 analogs as novel Fabry disease biomarkers. These molecules correspond to lyso-Gb3 with different chemical modifications on the sphingosine chain (-C2 H4 , -H2 , +O, +H2 O, +H2 O2, and +H2 O3 ). An ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the multiplex analysis of lyso-Gb3 and its 6 analogs in plasma. The samples are prepared by solid phase extraction using mixed-mode strong cation exchange (MCX) cartridges. An in-house synthesized N-glycinated lyso-Gb3 derivative was used for the internal standard. The limits of detection (LODs) measured for lyso-Gb3 and its analogs ranged from 0.06 to 0.29 nM. © 2016 by John Wiley & Sons, Inc.

Keywords: Fabry disease; Lyso-Gb3; MS/MS; analogs; biomarkers; plasma; tandem mass spectrometry; ultra-performance liquid chromatography (UPLC).

Publication types

  • Validation Study

MeSH terms

  • Chromatography, High Pressure Liquid
  • Fabry Disease / blood*
  • Glycolipids / blood*
  • Glycolipids / chemistry*
  • Humans
  • Limit of Detection
  • Male
  • Reference Standards
  • Solid Phase Extraction
  • Sphingolipids / blood*
  • Sphingolipids / chemistry*
  • Tandem Mass Spectrometry*

Substances

  • Glycolipids
  • Sphingolipids
  • globotriaosyl lysosphingolipid

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