A variety of software is used to determine quantitative parameters from radionuclide imaging procedures. Knowledge of the variability of parameter values found in different hospitals is an important aspect of clinical audit of these techniques. This study investigated the variation in relative renal function measurement from static DMSA scintigraphy in the UK. Ten studies representing a range of ages and relative function values were distributed in digital form to 100 hospitals with the assistance of the gamma camera computer suppliers and regional audit coordinators. The studies were analysed at each participating hospital and details of the different techniques and computer systems used were documented. The median value of relative percentage function was assessed for each of the studies. Methods varied in terms of the view used for analysis (54% geometric mean, 46% posterior), the type of background subtraction (single region 32%, separate regions 60%, none 8%) and the definition of the renal regions of interest (73% manual, 27% semi-automatic). Eighty-eight percent of results were within two percentage points of the study median and 98% within five percentage points. There were statistically significant differences observed in the results arising from the view used for the analysis and the background subtraction protocol. The results indicate that relative renal function assessment from static DMSA scintigraphy in the UK is essentially a reliable procedure, although improvements could be made by standardizing the technique used.