The present study examines the potential of using the near-surface chlorophyll a concentration ([Chla]surf), as it can be derived from ocean color observation, to infer the column-integrated phytoplankton biomass, its vertical distribution, and ultimately the community composition. Within this context, a large High-Performance Liquid Chromatography (HPLC) pigment database was analyzed. It includes 2419 vertical pigment profiles, sampled in case 1 waters with various trophic states (0.03–6 mg Chla m−3). The relationships between [Chla]surf and the chlorophyll a vertical distribution, as previously derived by Morel and Berthon (1989), are fully confirmed. This agreement makes it possible to go further and to examine if similar relationships between [Chla]surf and the phytoplankton assemblage composition along the vertical can be derived. Thanks to the detailed pigment composition, and use of specific pigment biomarkers, the contribution to the local chlorophyll a concentration of three phytoplankton groups can be assessed. With some cautions, these groups coincide with three size classes, i.e., microplankton, nanoplankton and picoplankton. Corroborating previous regional findings (e.g., large species dominate in eutrophic environments, whereas tiny phytoplankton prevail in oligotrophic zones), the present results lead to an empirical parameterization applicable to most oceanic waters. The predictive skill of this parameterization is satisfactorily tested on a separate data set. With such a tool, the vertical chlorophyll a profiles of each group can be inferred solely from the knowledge of [Chla]surf. By combining this tool with satellite ocean color data, it becomes possible to quantify on a global scale the phytoplankton biomass associated with each of the three algal assemblages.