Jump to ContentJump to Main Navigation

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Schlattmann, Peter / Tate, Jillian R. / Tsongalis, Gregory J.

12 Issues per year


IMPACT FACTOR 2014: 2.707
Rank 6 out of 30 in category Medical Laboratory Technology in the 2014 Thomson Reuters Journal Citation Report/Science Edition

SCImago Journal Rank (SJR): 0.860
Source Normalized Impact per Paper (SNIP): 1.046

VolumeIssuePage

Issues

Determination of globotriaosylceramide in plasma and urine by mass spectrometry

Ralf Krüger1 / Kai Bruns1, a / Silke Grünhage1 / Heidi Rossmann1 / Jörg Reinke2 / Michael Beck2 / Karl J. Lackner1

1Institute of Clinical Chemistry and Laboratory Medicine, Medical Center of the Johannes Gutenberg University, Mainz, Germany

2Department of Pediatrics/Villa Metabolica, Medical Center of the Johannes Gutenberg University, Mainz, Germany

aPresent address: Sanofi-Aventis Deutschland GmbH, Industriepark Hoechst, Frankfurt, Germany.

Corresponding author: Dr. Ralf Krüger, Institute of Clinical Chemistry and Laboratory Medicine, Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55131 Mainz, Germany

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 48, Issue 2, Pages 189–198, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2010.048, December 2009

Publication History

Received:
2009-07-10
Accepted:
2009-10-07
Published Online:
2009-12-04

Abstract

Background: Fabry disease is an X-chromosomally inherited lysosomal storage disorder leading to accumulation of glycosphingolipids, mainly globotriaosylceramide (ceramide-trihexoside, Gb3). Concentrations of Gb3 in plasma and urine have been used to diagnose Fabry disease and to monitor enzyme replacement therapy with recombinant α-galactosidase.

Methods: Gb3 was purified from plasma or urine by combined liquid extraction/protein precipitation and solid-phase extraction, and was detected by flow-injection analysis electrospray mass spectrometry (MS) using multi-reaction-monitoring. Calibration was performed via standard addition using C17-Gb3 as internal standard. The most abundant isoforms were monitored for calculation of total Gb3.

Results: A MS-based assay for quantification of Gb3 in plasma and urine was established and validated. Intra- and inter-assay coefficient of variation (CV) of the method were ≤12%. However, at low concentrations the CV was 16%. The linear range covers roughly two orders of magnitude, down to 0.54 mg/L in plasma and 0.07 mg/L in urine. Careful adjustment of tuning parameters was necessary to obtain identical isoform intensities and quantitative results on different mass spectrometers. Gb3 concentrations in healthy controls were <4 mg/L in EDTA-plasma and <10 μg/mmol creatinine in urine. Significantly increased Gb3 concentrations were found in plasma and urine from male and female patients with Fabry disease.

Conclusions: An improved MS protocol for Gb3 quantification has been developed, validated, and shown to be suitable for diagnosis and monitoring of Fabry patients.

Clin Chem Lab Med 2010;48:189–98.

Keywords: Fabry disease; Gb3 (globotriaosylceramide); glycosphingolipid; liquid chromatography-tandem mass spectrometry; lysosomal storage disorder

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Sandra Merscher and Alessia Fornoni
Frontiers in Endocrinology, 2014, Volume 5
[2]
Hany Farwanah and Thomas Kolter
Metabolites, 2012, Volume 2, Number 4, Page 134
[3]
J. Lukas, J. Torras, I. Navarro, A.-K. Giese, T. Bottcher, H. Mascher, K. J. Lackner, G. Fauler, E. Paschke, J. M. Cruzado, A. Dudesek, M. Wittstock, W. Meyer, and A. Rolfs
Clinical Kidney Journal, 2012, Volume 5, Number 5, Page 395
[4]
D. Ryan, K. Robards, P.D. Prenzler, and Megan Kendall
Analytica Chimica Acta, 2011, Volume 684, Number 1-2, Page 17
[5]
C. Tanislav, M. Kaps, A. Rolfs, T. Böttcher, K. Lackner, E. Paschke, H. Mascher, M. Laue, and F. Blaes
European Journal of Neurology, 2011, Volume 18, Number 4, Page 631
[6]
Journal of Mass Spectrometry, 2010, Volume 45, Number 7, Page 829

Comments (0)

Please log in or register to comment.