A method has been developed in which the concentrations of insulin in dried blood spots on filter paper can be estimated. The technique involves elution of blood from the paper, evaporation, and rehydration in assay buffer. The resuspended samples are then assayed by radioimmunoassay. The ratio of control plasma to control dried blood spot insulin values is used as a conversion factor for unknown samples on paper to express the dried blood spot insulin concentration in plasma equivalents. Intra-assay variability was 14%, and interassay variability was 25%, but the elution conversion factor had a variability of about 40% among individuals. This technique has substantial advantages for field studies, but elevated values should be confirmed by a venous plasma sample measurement in a conventional radioimmunoassay.