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The aim of this study was to develop a simplified method for quantitative analysis of liver scintigraphy with 99Tcm-diethylenetriamine pentaacetic acid-galactosyl-human serum albumin (GSA) using spectral analysis. Dynamic liver scintigraphy using GSA was performed in three normal volunteers and 19 patients with chronic liver disease. Dynamic data were obtained with a gamma camera for 30 min after the injection of approximately 185 MBq GSA. The rate constant for the liver uptake of GSA from the blood (Ku, min-1), total excretion rate (Ke, min-1) and non-specific volume of distribution (Vh) were obtained by spectral analysis. Vh was defined as the volume in the liver region of interest (ROI) occupied by GSA which was in equilibrium with that in the blood. It should be noted that Vh had no units, since the counts in both the liver and heart ROIs were normalized by scan length to obtain counts pixel-1 min-1. For comparison, compartmental analysis was also performed. A receptor index (LHL15) was calculated by dividing the radioactivity of the liver ROI by that of the liver plus heart ROIs 15 min post-injection. The Ku values obtained by spectral analysis (y) agreed well with those obtained by compartmental analysis (x) (y = 0.953x - 0.013, r = 0.992, S.E.E. = 0.016 min-1). The Ke and Vh values obtained by spectral analysis (y) correlated significantly with those obtained by compartmental analysis (x) (y = 1.149x - 0.016, r = 0.826, S.E.E. = 0.017 min-1 for Ke; y = 1.191x + 0.044, r = 0.975, S.E.E. = 0.021 for Vh). The Ku values obtained by spectral analysis decreased as the severity of liver disease progressed, and were non-linearly related to the LHL15 values, suggesting that Ku is more sensitive to liver damage than LHL15, especially in the early stages of liver damage. These results suggest that spectral analysis applied to dynamic liver scintigraphy with GSA provides a simple, non-invasive and useful tool for the quantitative evaluation of liver function.