This study was performed to evaluate the usefulness of spectral analysis (SA) applied to dynamic single photon emission computed tomography studies with N-isopropyl-p-(123I)iodoamphetamine (IMP). The unidirectional clearance of IMP from the blood to the brain tissue (K1) obtained by SA (y (ml/g/min)) agreed well with that obtained from a two-compartment model using the nonlinear least-squares (NLSQ) method (x (ml/g/min)) (y = 0.994x + 0.003, r = 0.999, standard error of the estimate (SEE) = 0.005 ml/g/min). The rate constant for back diffusion of IMP from the brain tissue to the blood (k2) obtained by SA (y (min(-1))) also agreed well with that obtained by the NLSQ method (x (min(-1))) (y = 0.985x + 0.000, r = 0.948, SEE = 0.001 min(-1)). The brain vascular volume (V0) obtained by SA (y (ml/g)) correlated well with that obtained by the NLSQ method (x (ml/g)) (y = 1.138x + 0.000, r = 0.867, SEE = 0.012 ml/g). These results indicate that SA is applicable and useful for quantification of the kinetic parameters of IMP in the human brain, and can be an alternative approach to compartment analysis.