A new multiplex analysis of glucosylsphingosine and globotriaosylsphingosine in dried blood spots by tandem mass spectrometry

Amber Van Baelen; Laurence Roosens; Sylvie Devos; Stijn Verhulst; François Eyskens

doi:10.1016/j.ymgmr.2023.100993

A new multiplex analysis of glucosylsphingosine and globotriaosylsphingosine in dried blood spots by tandem mass spectrometry

Mol Genet Metab Rep. 2023 Aug 18:37:100993. doi: 10.1016/j.ymgmr.2023.100993. eCollection 2023 Dec.

Authors

Amber Van Baelen^{1

2

3}, Laurence Roosens⁴, Sylvie Devos⁴, Stijn Verhulst^{2

3}, François Eyskens^{1

2

3}

Affiliations

¹ Center of Inherited Metabolic Diseases, UZA, Antwerp, Belgium.
² Laboratory of Experimental Medicine and Pediatrics, University of Antwerp, Belgium.
³ Pediatric Department, UZA, Antwerp, Belgium.
⁴ Metabolic lab CBAA, UZA, Antwerp, Belgium.

Abstract

Background: Gaucher's and Fabry's disease are two of the most common treatable lysosomal storage diseases, and have a wide spectrum of clinical symptoms. Early detection is important, because timely initiation of treatments can improve the disease status and prevent complications. However disease manifestations develop in childhood, diagnosis is delayed until adulthood partly due to the limitations of the currently used diagnostic pathway. The aim of this research is to develop and validate a multiplex assay and defining reference ranges, which do not exist at this moment, to improve and facilitate the entire diagnostic work up and enable treatment in an earlier stage of disease.

Methods and findings: Biomarkers glucosylsphingosine (GlcSph) and globotriaosylsphingosine (Lyso-Gb3) were detected and quantified using LC-MS/MS on dried blood spots. We developed an improved and new extraction method that allowed to measure GlcSph and Lyso-Gb3 in a multiplex analytical platform. After validation of the method, samples of 1480 individuals with normal enzymatic activity were collected to determine age and gender-related reference ranges.Our combination method showed a good linearity, precision, accuracy and limit of quantification with lack of carry-over following the specific international CLSI guidelines. The suggested protocol is robust, efficient, sensitive, specific, comprehensive and relatively cheap in order to accelerate the diagnostic process for both lysosomal storage diseases. The samples, with normal enzymatic activity, defined statistical relevant and clinical correct reference ranges for each specific age group by gender.

Conclusion: We report a multiplex LC-MS/MS method and relevant reference ranges that are appropriate for the targeted screening, diagnosis and follow-up of Fabry and Gaucher disease.

Keywords: Biomarker; Dried blood spots; Globotriaosylsphingosine; Glucosylsphingosine; Lysosomal storage diseases; Reference ranges; Tandem mass spectrometry.