Purpose : The aim of this study was to assess the validity of standard deviation of gray scale histogram in digital subtraction radiography as a test parameter for superimposition error. Materials and Methods : Twenty periapical radiographs were used as baseline images and they were copied to exclude the influence of exposure geomety and contrast differences. These subsequent images were linearly displaced by 0.1-0.5 mm in the x-. y- and xy-directions, rotated by 0.5-3° and distorted by angular contraction of 1-5° in x- and y-axis before subtraction. The standard deviations of gray levels in the subtraction images were obtained and paired t-tests were performed. Pearson correlation coefficients(r) were calculated between the standard deviations and the superimposition errors. Results : Linear displacement showed high correlation coefficients of 0.997, 0.997 and 0.995 in x-. y- and xy-axis respectively. Statistically significant different standard deviation existed among all linearly displaced groups(p<0.05). Distortion showed relatively low correlation coefficients of 0.982 and 0.959 in x- and y-axis. The standard deviations between the two distortion groups were statistically significant different(p<0.05). Conclusion : Standard deviation of gray level distribution in digital subtraction images is satisfactory but not perfect similarity measure to assess the superimposition errors.