The late Pleistocene to early Holocene Champlain Sea provides a unique opportunity to study the development of marine ecosystems in a context of global climatic change. This study presents radiocarbon (14C) dates and stable isotope analyses on 15 vertebrate specimens from Champlain Sea sediments, including the Charlotte Whale, which is Vermont’s State marine fossil. Data are used in an attempt to investigate the timing of colonization and ecological dynamics in this newly formed sea. Using the average marine correction, 14C dates on four specimens likely calibrate prior to or possibly synchronous with the accepted origination date for the Champlain Sea, implying larger marine reservoir effects than the average marine correction in the vertebrate tissues. Without knowing the specific marine reservoir offsets, it is not possible to calculate the timing of colonization or its relation to concurrent climatic change. Observed lower δ13C and δ15N values in walruses, a fin whale, and a right whale support consumption of prey from lower trophic levels such as bivalve mollusks, krill, and copepods. Higher isotopic values in beluga whales and a bird, the thick-billed murre, support consuming fish, such as cod and capelin. These isotopic data show comparable values and relationships as observed in modern arctic marine ecosystems.