D-Mannose-1-¹³C is widely used as a stable isotopically labeled tracer in metabolic studies, particularly in glycobiology and cancer research. Its primary application lies in tracking the metabolic fate of mannose in cellular pathways, including glycosylation processes and the biosynthesis of glycoproteins and glycolipids. Due to the ¹³C label at the first carbon position, it enables precise monitoring using NMR spectroscopy and mass spectrometry, allowing researchers to distinguish between endogenous and exogenous mannose pools.

It is also employed in studies of congenital disorders of glycosylation (CDG), where altered sugar metabolism affects protein function. In microbiology, D-Mannose-1-¹³C helps investigate bacterial cell wall synthesis, especially in pathogens that utilize mannose-containing polysaccharides. Additionally, it supports research into mannose metabolism in immune cells, providing insights into immunomodulation and host-pathogen interactions. The compound is valuable in flux analysis to quantify pathway activities in live cells and tissues.