Testosterone plays multiple roles in the regulation of development, physiology, reproduction, and behavior. Age-related testosterone declines are expected in the population. However, measuring circulating testosterone is especially challenging because concentrations are labile, responding to social situations and challenges. Matrices that integrate long-term testosterone levels are therefore valuable as biomarkers of endogenous levels as well as chronic exposures. Here, we report on a simple method to extract and measure accumulated testosterone from human fingernails using commercial enzyme immunoassay kits. Furthermore, we demonstrate known human testosterone sex and age trends. Our method is especially useful for quantifying testosterone in men's nails, where a small amount of matrix is required. Thus, this approach is a potential tool for biomonitoring endogenous as well as exogenous testosterone exposure. We suggest considering nails as an alternative matrix for quantifying other steroids as well.