Heavy metals, including cadmium, are common contaminants in environments subject to human activity. Responses to exposure in the fruit fly, Drosophila melanogaster, are dosage-dependent and resistance is selectable. While metallothionein-mediated sequestration has been extensively studied as a mechanism of cadmium resistance, a link between selection for resistance and an increased accumulation of cadmium has yet to be demonstrated. To address this need, we have selected wild-type flies for cadmium resistance for 20 generations and tested metal content using mass spectrometry. Resistant flies were observed to contain lower levels of cadmium, arguing for a mechanism of cadmium resistance that is not mediated by increased sequestration. This, coupled with genetic evidence suggesting the involvement of factors located on the X chromosome, suggests a gene other than metallothionein may be involved in resistance in this line.