Simultaneous quantification of sphingolipid metabolites in human plasma using UPLC-MS/MS.

Area of Science:

• Biochemistry
• Metabolomics
• Hepatology

Background:

• Sphingolipids, including sphingosine (SP), dihydro-sphingosine (DSP), and sphingosine 1-phosphate (S1P), play critical roles in metabolic-associated fatty liver disease (MAFLD).
• Accurate quantification of these metabolites is essential for understanding their involvement in MAFLD pathogenesis.

Purpose of the Study:

• To develop and validate a sensitive and specific analytical method for the simultaneous quantification of SP, DSP, and S1P in human plasma.
• To investigate the levels of these sphingolipids in patients with MAFLD compared to healthy controls.

Main Methods:

• Ultra-high-performance liquid chromatography (UPLC) coupled with tandem mass spectrometry (MS/MS) was employed for quantification.
• A stripped biological matrix using activated-carbon treatment and protein precipitation with methanol were used for sample preparation.
• Detection was performed using multiple reaction monitoring (MRM) on a triple quadrupole mass spectrometer.

Main Results:

• The developed UPLC-MS/MS method demonstrated excellent selectivity, linearity, accuracy, precision, recovery, matrix effects, and stability.
• Plasma concentrations of SP, DSP, and S1P were significantly elevated in MAFLD patients (n=30) compared to healthy controls (n=30).

Conclusions:

• The validated UPLC-MS/MS method provides a reliable tool for measuring key sphingolipids in plasma.
• Elevated levels of SP, DSP, and S1P are associated with MAFLD, suggesting their potential role as biomarkers or therapeutic targets.