To date, environmental conditions have been enough to act as an effective barrier to prevent non-indigenous species from arriving and establishing in Arctic Canada. However, rapidly changing climatic conditions are creating more suitable habitats for non-indigenous species to potentially establish and become invasive. Concurrently, shipping traffic in parts of Arctic Canada has increased by over 250% since 1990, providing an effective vector for transporting non-indigenous species to the region. Arctic Canada has been historically undersampled, so Arctic biota inventories are incomplete, hampering efforts to establish if a species is new to the region (and potentially invasive) or newly discovered. In this study, we utilize environmental DNA (eDNA) metabarcoding and ships of opportunity to assess eukaryotic community composition and potential invasives along one of the busiest shipping routes, the Northwest Passage. One liter seawater samples were collected in triplicate at 27 locations, targeting touristic hotspots frequently visited by passenger vessels. Eukaryotic DNA was amplified from the 18S rRNA V9 and COI regions, resulting in 126 unique Amplicon Sequence Variants (ASVs) detected with COI and 391 ASVs with 18S, providing an important snapshot of current community composition. Copepods, dinoflagellates, and diatoms were the most abundant taxonomic groups, correlating well with previous net sampler surveys, validating the efficacy of eDNA for biodiversity surveillance. We also report the first detections of a prolific invasive species, the bay barnacle (Amphibalanus improvisus), in Arctic Canada. Further work is currently in progress to establish whether these detections represent transient barnacle larvae or sessile adults capable of recruiting and reproducing. Our study demonstrates the utility of eDNA for the detection of non-indigenous species in a data-poor area, which, if combined with citizen science initiatives and local communities, could provide a vital monitoring tool for the detection of new invasives in this rapidly changing area.