J Genet Med.  2007 Jun;4(1):45-52.

Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease

  • 1Department of Biomedical & Pharmaceutical Analysis, College of Pharmacy, Duksung Women's University, Seoul, Korea. hyeran11@duksung.ac.kr
  • 2Department of Analytical Research, BioCore Co., LTD, Korea.
  • 3Department of Pediatrics, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.
  • 4Department of Pediatrics, College of Medicine, Chungnam National University, Daejeon, Korea.
  • 5Department of Pediatrics, Soonchonhyang University College of Medicine, Korea.
  • 6Therapeutic Protein Research, Genzyme Co., Farmingham, USA.


A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-extensive pre-treatment by electrospray ionization MS/MS (ESI-MS/MS). Measurement of globotriaosylceramide (Gb3, ceramide trihexoside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ??-galactosidase A (alpha-Gal A). The lack of alpha-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3.
Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS.
Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms was found in the range of 0.001-1.0 microgram/mL. The limit of detection (S/N=3) was 0.001 microgram/mL and limit of quantification was 0.01 microgram/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982.
This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.


Globotriaosylceramide (Gb3); MS/MS; Fabry disease; Quantification; diagnosis
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