This report describes the analysis of 20 human scalp hairs, and one facial hair, found attached to bristles of a hairbrush recovered during underwater archaeological investigation of the wreck of HMS Erebus of the British Royal Navy’s “Franklin Expedition” which sank in the Arctic after its desertion in April 1848. Hair colour and diameter range were typical of Western Europeans, consistent with the Caucasian origins of the crew. Microscopic examination showed often severe degradation caused by biological agents and mechanical damage, and penetrative contamination by iron oxide encrustation from the wreck environment during some 170 years of marine immersion. No viable DNA was recovered to allow identification from a database of crew descendants. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) determined heavy and trace metals within the hair that might reflect uptake or depletion of those metals during life to allow insight to conditions on board, or contamination during immersion. Further analysis of hair from a present-day male group and from two Franklin Expedition crewmen whose permafrost-preserved remains were autopsied in the 1980s, and hog-hair bristles of the hairbrush, allowed some account of differential effects of time upon the hair structure while immersed or non-immersed. Analysis confirmed considerable contamination of the hair from metals likely to derive from the wreck and marine environment (e.g., As, Hg, Na, Pb, etc.). The large range of concentrations found in the hair from Erebus precluded any insight to the health status or otherwise of the donor(s) of the hairs. Conclusions were further limited by uncertainty over whether the hairs were from only one individual and the time at which the hairs were deposited on the brush, for example before and/or during the mission, and at what stage in the known chronology of events the last hairs were deposited. Nonetheless, the present retrieval and analysis of human hair from this iconic wreck may provide a further advance in the archaeological investigation of the Franklin Expedition.