Development of a new online SPE-HPLC-MS/MS method for the profiling and quantification of sphingolipids and phospholipids in red blood cells – Application to the study of Gaucher's disease

• Published in: Analytica chimica acta Vol. 1278; p. 341719
• Main Authors: Bettioui, Terkia, Chipeaux, Caroline, Ben Arfa, Kaouther, Héron, Sylvie, Belmatoug, Nadia, Franco, Mélanie, de Person, Marine, Moussa, Fathi
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 16.10.2023; Elsevier Masson
• Subjects: Chemical Sciences; Gaucher's disease; Online SPE-HPLC-MS/MS; Red blood cells lipidome; Sphingolipid and phospholipid profiling; Red blood cells lipidome; Online SPE-HPLC-MS/MS; Sphingolipid and phospholipid profiling; Gaucher's disease
• ISSN: 0003-2670; 1873-4324; 1873-4324
• DOI: 10.1016/j.aca.2023.341719

Red blood cells (RBCs) are the subject of clinical attention due to their biological importance. Recently, it has been shown that certain erythrocyte pathologies could be linked to an abnormal lipid composition. In this work, we have developed a simple and fast method using online sample preparation with liquid chromatography coupled to mass spectrometry (SPE-HPLC-MS/MS), to identify a large number of sphingolipids (SL) and phospholipids (PL). The use of online sample preparation considerably reduces analysis times (15 min including extraction and separation of lipids + 2 min for system re-equilibration) and facilitates experimentation while ensuring very good extraction yields. This method was then successfully applied to the quantification of 30 sphingolipids and phospholipids in plasma and erythrocyte extracts from a cohort of individuals with Gaucher disease, treated or not by enzymotherapy. Our results for the study of this disease, led us to establish the lipid profile of the healthy red blood cells, still not very well-known to date. For this, we adopted a semi-targeted approach, based on the use of a triple-quadrupole analyzer and identified more than two hundred different lipid species. These promising results will hopefully enable us to enrich our knowledge of the normal red blood cells lipidome. [Display omitted] •Optimization and validation of an online sample preparation HPLC-MS/MS methodology for the dosage of sphingolipids and phospholipids in plasma and red blood cells (RBC).•Comparison between online sample preparation and LLE performances.•Application to the study of Gaucher's disease: search of new biomarkers and evaluation of ERT therapy on the lipid profile.•Exploratory study of the RBC lipidome by adopting a semi-targeted approach.