The aim of this study was to develop a new algorithm for volume determination by single photon emission computed tomography (SPECT). Different algorithms were evaluated through phantom studies. The results show that the algorithm combining moment-preserving bilevel thresholding and best-fit Laplacian second derivative edge detection can provide the most accurate measurement of volume. Besides, this method can be utilized in different SPECT systems with no need for further phantom studies. In patient studies, the results of liver volume calculation have indicated that this is a useful technique in clinical medicine.