The main purpose of the study was to compare proton (1H) single-voxel MR spectroscopy (MRS) with high-spatial-resolution spectroscopic imaging (MRSI) to determine the lipid content in human skeletal muscle. Unsuppressed water line was used as a concentration reference in the processing of single-voxel spectra. The spectrum from yellow bone marrow with a 100% fat content and probe with the vegetable oil served as internal and external reference for high-spatial-resolution MRSI, respectively. Very good correlation was found between lipid concentrations measured by water referenced single-voxel MRS and high-spatial-resolution MRSI with yellow bone marrow as the internal standard. Excellent correlation was found between total lipid concentrations estimated by high-spatial-resolution MRSI with vegetable oil as the external fat standard and yellow bone marrow as the internal reference. From comparison of single-voxel MRS and MRSI approaches, it follows that relaxation correction of the reference water and methylene fat line is inevitable in processing the standard single-voxel spectra. The high-resolution MRSI approach is recommended to avoid the problem of relaxation corrections and enables using vegetable oil as the external fat standard.