Biomass burning impacts biogeochemical cycling, vegetation dynamics and climate. However, interactions between fire, climate and vegetation are not well understood and therefore studies have attempted to reconstruct fire and vegetation history under different climatic conditions using sedimentary archives. Here we focus on levoglucosan, a thermal by-product of cellulose generated during biomass burning, and, therefore, a potential fire biomarker in the marine sedimentary archive. However, before levoglucosan can be applied as a biomass burning proxy in marine sediments, there is a need for studies on how levoglucosan is transported to the marine environment, how it is reflecting biomass burning on continents, as well as the fate of levoglucosan in the marine water column and during deposition in marine sediments. Here we present analyses of levoglucosan, using an improved Ultra High Pressure Liquid Chromatography-Electro Spray Ionization/High Resolution Mass Spectrometry (UHPLC-ESI/HRMS) method, in atmospheric particles, in particulate matter settling through the water column and in marine surface sediments on a longitudinal transect crossing the tropical North Atlantic Ocean at 12°N. Levoglucosan was detected in the atmosphere, although in low concentration, possibly due to the sampled particle size, the source area of the aerosols, or the short time interval of sampling by which large burning events may have been missed. In sinking particles in the tropical North Atlantic Ocean we find that levoglucosan deposition is influenced by a mineral ballast effect associated with marine biogenic particles, and that levoglucosan is not transported in association with mineral dust particles. Highest levoglucosan concentrations and seasonal differences in sinking particles were found close to continents and low concentrations and seasonal differences were found in the open ocean. Close to Africa, levoglucosan concentration is higher during winter, reflecting seasonal burning in northwestern Africa. However, close to South America levoglucosan concentrations appear to be affected by riverine transport from the Amazon River. In surface sediments close to South America, levoglucosan concentration is higher than in the middle of the Atlantic Ocean, implying that here the influence from the South American continent is important and perennial. Our study provides evidence that degradation of levoglucosan during settling in the marine water column is not substantial, but is substantial at the sediment–water interface. Nevertheless, levoglucosan was detected in all surface sediments throughout the tropical North Atlantic, indicating its presence in the marine sedimentary record, which reveals the potential for levoglucosan as a biomass burning proxy in marine sediments.