Quantification of Globotriaosylsphingosine in Plasma and Urine of Fabry Patients by Stable Isotope Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry
Open Access
- 1 March 2013
- journal article
- Published by Oxford University Press (OUP) in Clinical Chemistry
- Vol. 59 (3), 547-556
- https://doi.org/10.1373/clinchem.2012.192138
Abstract
BACKGROUND Biochemical markers that accurately reflect the severity and progression of disease in patients with Fabry disease and their response to treatment are urgently needed. Globotriaosylsphingosine, also called lysoglobotriaosylceramide (lysoGb3), is a promising candidate biomarker. METHODS We synthesized lysoGb3 and isotope-labeled [5,6,7,8,9] 13C5-lysoGb3 (internal standard). After addition of the internal standard to 25 μL plasma or 400 μL urine from patients with Fabry disease and healthy controls, samples were extracted with organic solvents and the lysoGb3 concentration was quantified by UPLC-ESI-MS/MS (ultraperformance liquid chromatography–electrospray ionization–tandem mass spectrometry). Calibration curves were constructed with control plasma and urine supplemented with lysoGb3. In addition to lysoGb3, lyso-ene-Gb3 was quantified. Quantification was achieved by multiple reaction monitoring of the transitions m/z 786.4 > 282.3 [M+H]+ for lysoGb3, m/z 791.4 > 287.3 [M+H]+ for [5,6,7,8,9] 13C5-lysoGb3, and 784.4 > 280.3 [M+H]+ for lyso-ene-Gb3. RESULTS The mean (SD) plasma lysoGb3 concentration from 10 classically affected Fabry hemizygotes was 94.4 (25.8) pmol/mL (range 52.7–136.8 pmol/mL), from 10 classically affected Fabry heterozygotes 9.6 (5.8) pmol/mL (range 4.1–23.5 pmol/mL), and from 20 healthy controls 0.4 (0.1) pmol/mL (range 0.3–0.5 pmol/mL). Lyso-ene-Gb3 concentrations were 10%–25% of total lysoGb3. The urine concentration of lysoGb3 was 40–480 times lower than in corresponding plasma samples. Lyso-ene-Gb3 concentrations in urine were comparable or even higher than the corresponding lysoGb3 concentrations. CONCLUSIONS This assay for the quantification of lysoGb3 and lyso-ene-Gb3 in human plasma and urine samples will be an important tool in the diagnosis of Fabry disease and for monitoring the effect of enzyme replacement therapy in patients with Fabry disease.Keywords
This publication has 14 references indexed in Scilit:
- Quantification of the Fabry marker lysoGb3 in human plasma by tandem mass spectrometryJournal of Chromatography B, 2012
- Reduction of elevated plasma globotriaosylsphingosine in patients with classic Fabry disease following enzyme replacement therapyBiochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2011
- How well does urinary lyso-Gb3 function as a biomarker in Fabry disease?Clinica Chimica Acta; International Journal of Clinical Chemistry, 2010
- Plasma globotriaosylsphingosine: Diagnostic value and relation to clinical manifestations of Fabry diseaseBiochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2010
- Tissue and plasma globotriaosylsphingosine could be a biomarker for assessing enzyme replacement therapy for Fabry diseaseBiochemical and Biophysical Research Communications, 2010
- Thematic Review Series: Sphingolipids. Biodiversity of sphingoid bases (“sphingosines”) and related amino alcoholsJournal of Lipid Research, 2008
- Elevated globotriaosylsphingosine is a hallmark of Fabry diseaseProceedings of the National Academy of Sciences of the United States of America, 2008
- Treatment of Fabry Disease: Outcome of a Comparative Trial with Agalsidase Alfa or Beta at a Dose of 0.2 mg/kgPLOS ONE, 2007
- The Dutch Fabry cohort: Diversity of clinical manifestations and Gb3 levelsJournal of Inherited Metabolic Disease, 2007
- A RAPID METHOD OF TOTAL LIPID EXTRACTION AND PURIFICATIONCanadian Journal of Biochemistry and Physiology, 1959