The characteristics of technetium-99m diethylenetriaminepentaacetic acid human serum albumin (99mTc-DTPA-HSA) as an agent for quantitation of cerebral blood volume (CBV) were examined. The radioactivity after decay correction as a percentage of the activity at 10 minutes was 84.3 +/- 1.3% at 120 minutes after the injection of 99mTc-DTPA-HSA. Radioactivity was found exclusively in plasma, with little in blood cells. The blood retention of 99mTc-DTPA-HSA is sufficient, and its use in the quantitation of CBV omits the need for centrifugation of the blood sample. CBV quantified using the tracer and a SPECT system with a single-head rotating gamma camera was 4.09 +/- 0.60 ml/100 g brain, similar to values reported previously. Two serial SPECT scans provided similar images, and the CBV values determined by the two scans were closely correlated (p < 0.001). The high reproducibility suggests that the method may be used to monitor the cerebrovascular response to drugs. It was concluded that 99mTc-DTPA-HSA has useful properties for quantitative CBV measurement and that quantitation of CBV by 99mTc-DTPA-HSA SPECT is feasible using a system with a single-head rotating gamma camera.