Humans are exposed to concentrations of multiple trace elements through a variety of background sources; many are suspected reproductive toxicants. Prior to investigating associations between trace elements and human reproductive health, potential biomarkers of exposure should be characterized by sources of variability in the population at risk. Factors influencing elemental exposure should also be identified to ensure their consideration as potential confounding variables. The principal aim of this study is to characterize sources of variability for 19 trace elements measured in urine specimens collected from 55 women and 36 male partners completing a 1st cycle of in vitro fertilization (IVF). Urine specimens were analyzed using a biomonitoring method based on inductively coupled plasma-mass spectrometry (ICP-MS). Randomly selected urine specimens (∼6%) were analyzed in duplicate, and these data were used to characterize sources of variability. Nine trace elements including As, Ba, Cd, Cs, Co, Cu, Mn, Mo, and Zn, were quantified in most specimens, indicating their utility in future epidemiologic studies of trace elements exposure and IVF outcomes. With few exceptions, normalizing urine using the traditional creatinine-correction procedure, or an alternative approach based on a linear regression model, increased residual variability only slightly. Sex and race appear to be important factors to consider in epidemiologic studies conducted in this population. Urine concentrations for most elements are similar to those reported in the 2005-2006 National Health and Nutrition Examination Survey (NHANES); however, differences in others may indicate regional trends or a unique exposure history for this infertile study population.