Intracranial diseases are common in dogs and improved noninvasive diagnostic tests are needed. Magnetic resonance (MR) spectroscopy is a technique used in conjunction with conventional MR imaging to characterize focal and diffuse pathology, especially in the brain. As with conventional MR imaging, there are numerous technical factors that must be considered to optimize image quality. This study was performed to develop an MR spectroscopy protocol for routine use in dogs undergoing MR imaging of the brain. Fifteen canine cadavers were used for protocol development. Technical factors evaluated included use of single-voxel or multivoxel acquisitions, manual placement of saturation bands, echo time (TE), phase- and frequency-encoding matrix size, radiofrequency coil, and placement of the volume of interest relative to the calvaria. Spectrum quality was found to be best when utilizing a multivoxel acquisition with the volume of interest placed entirely within the brain parenchyma without use of manually placed saturation bands, TE = 144 ms, and a quadrature extremity radiofrequency coil. An 18 × 18 phase- and frequency-encoding matrix size also proved optimal for image quality, specificity of voxel placement, and imaging time.