Simultaneous quantitation of sphingoid bases by UPLC-ESI-MS/MS with identical ¹³C-encoded internal standards

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Diagnostics

Background:

• Free sphingoid bases (lysosphingolipids) are elevated in various sphingolipidoses.
• Accurate quantification of sphingoid bases is essential for diagnosing these genetic disorders.
• Previous methods relied on UPLC-ESI-MS/MS with 13C-encoded internal standards.

Purpose of the Study:

• To describe the feasibility of a simultaneous quantitation method for sphingoid bases in plasma.
• To validate the sensitivity, linearity, and assay variation of the multiplex assay.
• To analyze plasma samples from Gaucher Disease and Fabry Disease patients.

Main Methods:

• Utilized UPLC-ESI-MS/MS for sphingoid base quantification.
• Employed a mixture of 13C-encoded internal standards for simultaneous measurement.
• Analyzed plasma samples from Gaucher Disease and Fabry Disease patients.
• Demonstrated the conversion of sphingolipids to sphingoid bases for subsequent quantification.

Main Results:

• Achieved excellent sensitivity and linearity for all examined sphingoid bases.
• Observed acceptable intra- and inter-assay variation (<10% average).
• Successfully analyzed plasma samples from Gaucher and Fabry disease patients.
• Obtained data consistent with previous findings for specific biomarkers.

Conclusions:

• The developed multiplex assay is a feasible and reliable method for simultaneous sphingoid base quantification in plasma.
• This approach facilitates the diagnosis and monitoring of sphingolipidoses.
• The method can be extended to analyze other sphingolipids in the same samples.