and objectives: Fabry
disease is an X-linked lysosomal storage disease caused by the deficiency of
α-galactosidase A, resulting in glycosphingolipids accumulation in body
fluids and different organs. Globotriaosylceramide (Gb3),
globotriaosylsphingosine (Lyso-Gb3) and their analogues have been identified
and quantified as biomarkers for the disease severity and treatment efficacy.
The current study aimed to develop HPLC-MS methods in order to identify and
quantify FD biomarkers.
and Methods: Human
Fabry patients’ plasma and urine samples were processed using solid phase
extraction. The samples were then analysed for levels of Lyso-Gb3 and its
analogues using HPLC-ESI-MS.
recovery was 90 % for urine and 70 % for plasma. Reverse phase-HPLCmethods
were optimised with an isocratic elution of (0.1 % formic acid/50 %
acetonitrile) and flow rate of 3 μL/min. A multiple reaction monitoring mode
MS method was optimised for identification and quantification of metabolites
showing limit of detection of 10fmoles Lyso-Gb3. Lyso-Gb3 and 7 analogues
were detected showing comparable fragments. These analogues vary from
Lyso-Gb3 due to a modification in the sphingosine moiety.
have established an HPLC-ESI-MS approach for analysis of Lyso-Gb3 and its
analogues. Pilot data shows low levels of these biomarkers are quantified in
urine and plasma.