The Brachialis (BR) is placed under the Biceps Brachii (BB) deep in the upper arm. Therefore, the detection of the corresponding surface Electromyogram (sEMG) is a complex task. The BR is an important elbow flexor, but it is usually not considered in the sEMG based force estimation process. The aim of this study was to attempt to separate the two sEMG activities of the BR and the BB by using a High Density sEMG (HD-sEMG) grid placed at the upper arm and Canonical Component Analysis (CCA) technique. For this purpose, we recorded sEMG signals from seven subjects with two 8 × 4 electrode grids placed over BB and BR. Four isometric voluntary contraction levels were recorded (5, 10, 30 and 50 %MVC) for 90° elbow angle. Then using CCA and image processing tools the sources of each muscle activity were separated. Finally, the corresponding sEMG signals were reconstructed using the remaining canonical components in order to retrieve the activity of the BB and the BR muscles.