Combined Analysis Of Plasma Or Serum Glucosylsphingosine And Globotriaosylsphingosine By Uplc-Ms/Ms

Purpose: To develop a method for the combined analysis of plasma and serum glucosylsphingosine (lyso-Gb(1)) and globotriaosylsphingosine (lyso-Gb(3)), biomarkers of Gaucher disease (GD) and Fabry disease (FD), respectively.Methods: Internal standards were added to 100 mu L of plasma/serum and glycosphingolipids: lyso-Gb(1), lyso-Gb(3), and galactosylsphingosine (GalSph) were extracted with dichloromethane/methanol and analyzed by UPLC-MS/MS. Samples from unaffected controls and patients with GD were first analyzed using a HILIC column to separate lyso-Gb(1) from its isomer, GalSph. Samples from patients with FD or GD were analyzed using a C18 column to measure lyso-Gb(3) and the hexosylsphingosine (HexSph: lyso-Gb(1) + GalSph) fraction in a single combined method.Results: Extraction efficiency was between 73% and 87% and day-to-day variability showed a relative standard error of < 7.5%. GalSph was determined to have minimal to no contribution to the HexSph fraction in samples from unaffected controls and patients with GD. Lyso-Gb(3) and HexSph measurements by the combined method were in good agreement with established methods, with no bias.Conclusions: HexSph and lyso-Gb(3) analysis by reversed-phase chromatography UPLC-MS/MS is a cost-effective, time-efficient approach for evaluating these glycosphingolipid biomarkers in patients with a suspected or confirmed diagnosis of GD and FD.