Lysosphingolipid Quantitation in Plasma and Dried-Blood Spots Using Targeted High-Resolution Mass Spectrometry

  • J Clin Lab Anal. 2025 Jan;39(1):e25131. doi: 10.1002/jcla.25131.
Franklin Ducatez  1  2 ,  Wladimir Mauhin  3 ,  Jules Ottaviani  1 ,  Thomas Plichet  1 ,  Carine Pilon  1 ,  Olivier Lidove  3 ,  Fréderic Barbey  4 ,  Régine Perrichot  5 ,  Sabrina Vergnaud  6 ,  Marc G Berger  7  8 ,  Juliette Berger  7  8 ,  Nadia Belmatoug  9 ,  Yann Nadjar  10 ,  Foudil Lamari  11 ,  Esther Noel  12 ,  Stéphane Marret  2 ,  Soumeya Bekri  1 ,  Abdellah Tebani  1
Affiliations
  • 1. Department of Metabolic Biochemistry, Referral Center for Lysosomal Diseases, Normandie Univ, UNIROUEN, AIMS, SysMedLab, CHU Rouen, Rouen, France.
  • 2. Department of Neonatal Pediatrics, Intensive Care, and Neuropediatrics, Referral Center for Lysosomal Diseases, Normandie Univ, UNIROUEN, INSERM U1245, CHU Rouen, Rouen, France.
  • 3. Department of Internal Medicine-Rheumatology, Referral Center for Lysosomal Diseases, filière G2M, GH Diaconesses, Croix Saint Simon Hospital, Paris, France.
  • 4. Department of Immunology, University of Lausanne and University Hospital of Lausanne, Switzerland.
  • 5. Service de Néphrologie, Centre Hospitalier de Bretagne Atlantique, Vannes, France.
  • 6. UF Maladies Héréditaires Enzymatiques Rares-CGD, Institut de Biologie et de Pathologies, CHU de Grenoble Alpes, Grenoble, France.
  • 7. CHU Clermont-Ferrand, Hopital Estaing, CRB-Auvergne, Referral Center for Lysosomal Diseases, Clermont-Ferrand, France.
  • 8. Université Clermont Auvergne, EA 7453 CHELTER, Referral Center for Lysosomal Diseases, Clermont-Ferrand, France.
  • 9. Referral Center for Lysosomal Diseases, AP-HP Nord, Beaujon Hospital, Paris Cité University, filière G2M, MetabERN, Paris Cité University, France.
  • 10. Neurology Department, Referral Center for Neurometabolic and Lysosomal Diseases, filière G2M, Hôpital Pitié-Salpêtrière, Paris, France.
  • 11. AP-HP, Sorbonne Université, DMU Biogem-Metabolic Biochemistry Department, Referral Center for Lysosomal Diseases, filière G2M, Neurometabolic and Neurodegenerative Unit - Hôpital Pitié-Salpêtrière, Paris, France.
  • 12. University Hospital of Strasbourg, Strasbourg, BP, France.
PMID: 39727194 DOI: 10.1002/jcla.25131
Abstract

Background: Sphingolipidoses are rare inherited metabolic diseases belonging to lysosomal diseases. Early and accurate diagnosis is crucial for effective management and treatment. In this study, we aimed to develop a robust method to accelerate the diagnosis of these sphingolipidoses using dried blood spots and plasma.

Method: We employed high-resolution mass spectrometry coupled with liquid chromatography (LC-HRMS) to analyze 6 lysosphingolipids (GlcSph/Psychosine, LysoGb3, LysoSM, LysoSM509, LysoGM1, and LysoGM2) on dried blood spots and plasma samples. The method was used to measure the lysosphingolipid levels in a group of 30 control subjects and 204 samples from patients with sphingolipidoses (61 dB and 143 plasma) including Fabry, Gaucher, GM2 Gangliodosis, Niemann-Pick type A/B, and Niemann-Pick type C.

Results: The developed multiplex LC-HRMS method demonstrated linearity, precision, and quantification performances particularly for GlcSph/Psychosine and LysoGb3 on samples including controls and patients with sphingolipidoses. LysoSM showed recovery variability, wherease LysoGM1 and LysoGM2 showed higher matrix effect.

Conclusion: Our study presents a high-resolution mass spectrometry method along with the established cutoff values, providing a valuable tool for targeted screening, accurate diagnosis, and monitoring sphingolipidoses. Furthermore, DBS showed reliable results that lay the path to a broader adoption for screening these diseases.

Keywords
dried blood spot; lysosomal diseases; mass spectrometry; screening; sphingolipids.
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