Development and evaluation of a simultaneous strategy for pyrimidine metabolome quantification in multiple biological...

Area of Science:

• Biochemistry
• Metabolomics
• Analytical Chemistry

Background:

• Pyrimidines are essential biomolecules involved in nucleic acid synthesis and metabolic pathways.
• Understanding pyrimidine metabolism is crucial for various biological processes.

Purpose of the Study:

• To propose and establish the concept of the pyrimidine metabolome.
• To develop an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for analyzing pyrimidine metabolites.
• To investigate the dynamic balance and transient distribution of pyrimidines in biological samples.

Main Methods:

• Development of a novel UPLC-MS/MS analytical strategy.
• Quantification of 14 analytes across pyrimidine de novo and salvage pathways.
• Validation of method sensitivity (LLOQs < 10 ng/mL), precision (<15% deviation), and stability.
• Metabolomics analysis of plasma and brain samples across different species.

Main Results:

• The UPLC-MS/MS method demonstrated high sensitivity, precision, and stability.
• Metabolomics analysis revealed significant interspecies diversity in pyrimidine metabolism in plasma and brain.
• Pyrimidine biosynthetic metabolism was found to be highly active in the brain.
• Distinct pyrimidine metabolic profiles were observed in cells with metabolic disorders during proliferation and apoptosis.

Conclusions:

• The pyrimidine metabolome concept and the developed UPLC-MS/MS method provide a powerful tool for studying pyrimidine metabolism.
• The findings offer valuable reference data for pyrimidine metabolite concentrations in various bio-samples.
• This research advances our understanding of pyrimidine metabolic dynamics and its implications in health and disease.