<p>We examined multiple natural abundance isotope ratios of human hair to assess biological variability within and between geographic locations and, further, to determine how well these isotope values predict location of origin. Sampling locations feature differing seasonality and mobile populations as a robust test of the method. Serially-sampled hair from Cambridge, MA, USA, shows lower δ<sup>2</sup>H and δ<sup>18</sup>O variability over a one-year time course than model-predicted precipitation isotope ratios, but exhibits considerable differences between individuals. Along a ∼13° north-south transect in the eastern USA (Brookline, MA, 42.3 ° N, College Park, MD, 39.0 ° N, and Gainesville, FL, 29.7 ° N) δ<sup>18</sup>O in human hair shows relatively greater differences and tracks changes in drinking water isotope ratios more sensitively than δ<sup>2</sup>H. Determining the domicile of humans using isotope ratios of hair can be confounded by differing variability in hair δ<sup>18</sup>O and δ<sup>2</sup>H between locations, differential incorporation of H and O into this protein and, in some cases, by tap water δ<sup>18</sup>O and δ<sup>2</sup>H that differ significantly from predicted precipitation values. With these caveats, randomly chosen people in Florida are separated from those in the two more northerly sites on the basis of the natural abundance isotopes of carbon, nitrogen, hydrogen, and oxygen.</p