This study aimed to investigate the accuracy of estimating the volume of limb muscles (MV) using ultrasonographic muscle thickness (MT) measurements. The MT and MV of each of elbow flexors and extensors, knee extensors and ankle plantar flexors were determined from a single ultrasonographic image and multiple magnetic resonance imaging (MRI) scans, respectively, in 27 healthy men (23-40 years of age) who were allocated to validation ( n=14) and cross-validation groups ( n=13). In the validation group, simple and multiple regression equations using MT and a set of MT and limb length, respectively, as independent variables were derived to estimate the MV measured by MRI. However, only the multiple regression equations were cross-validated, and so the prediction equations with r(2) of 0.787-0.884 and the standard error of estimate of 22.1 cm(3) (7.3%) for the elbow flexors to 198.5 cm(3) (11.1%) for the knee extensors were developed using the pooled data. This approach did not induce significant systematic error in any muscle group, with no significant difference in the accuracy of estimating MV between muscle groups. In the multiple regression equations, the relative contribution of MT for predicting MV varied from 41.9% for the knee extensors to 70.4% for the elbow flexors. Thus, ultrasonographic MT measurement was a good predictor of MV when combined with limb length. For predicting MV, however, the unsuitability of a simple equation using MT only and the difference between muscle groups in the relative contribution of MT in multiple regression equations indicated a need for further research on the limb site selected and muscle analyzed for MT measurement.