A LC-MS/MS method to simultaneously profile 14 free monosaccharides in biofluids

Area of Science:

• Biochemistry and Metabolism
• Analytical Chemistry
• Clinical Diagnostics

Background:

• Monosaccharides play crucial roles in cellular metabolism and serve as potential biomarkers.
• Accurate quantification of monosaccharides is essential for both basic research and clinical applications.
• Existing methods may lack the simultaneous quantification capabilities needed for comprehensive analysis.

Purpose of the Study:

• To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for simultaneous quantification of 14 free monosaccharides.
• To evaluate the assay's performance according to established clinical guidelines.
• To apply the validated assay for monosaccharide profiling in cell culture media and human serum.

Main Methods:

• Development of a novel LC-MS/MS method for simultaneous separation and quantification of 14 free monosaccharides.
• Performance evaluation including linearity (R square > 0.995) and precision (coefficient of variation < 20% at LLOQ).
• Application of the assay to analyze monosaccharide profiles in conditioned media from LO2 and HEK293 cells, and in serum samples from glucose tolerance tests.

Main Results:

• The LC-MS/MS assay achieved simultaneous quantification of 14 monosaccharides within a 6-minute run time.
• Cell culture experiments demonstrated differential monosaccharide consumption and production between LO2 and HEK293 cells.
• Analysis of patient sera showed a significant increase in D-glucose post-glucose intake, with other monosaccharides remaining largely unaffected.

Conclusions:

• The developed LC-MS/MS assay meets clinical requirements for accurate and simultaneous monosaccharide profiling.
• The assay provides insights into distinct monosaccharide metabolism in liver (LO2) and kidney (HEK293) cells.
• Dietary intake has a minimal impact on the measured levels of six serum monosaccharides during a glucose tolerance test.