Purpose: The present study examines the efficacy of Creatine (Cr) levels in the determination of tumor grade on single-voxel (SV) and multi-voxel (MV) magnetic resonance spectroscopy (MRS) in patients with glial tumors. Material and Methods: A retrospective review was made of 20 SV and 20 MV MRS PDF images of patients with a pathological diagnosis of glial tumor. Cr values and metabolite ratios were measured in the tumor and compared with those in the healthy symmetrical brain parenchyma of the contralateral hemisphere. Results: A significant difference was noted in the (Cho+Cr)/NAA and Cho/NAA ratios on SV MRS between the high-grade tumors and the healthy contralateral hemisphere. On MV MRS, Cho/NAA ratios within the minimum and maximum Cr(h) voxels (Cho/NAAmin-Cr(h), Cho/NAAmax-Cr(h), respectively) were higher in high-grade tumors compared to healthy tissue. ROC analysis showed that the max-Cr(h) metabolite on MV MRS was successful in distinguishing low-grade tumors from high-grade tumors. Conclusion: Using the minimum and maximum values of Cr as a reference can improve the overall diagnostic accuracy in the diagnosis and grading of glial tumors. Our findings showed that Cr tended to be low in high-grade tumors and further that the max-Cr(h) metabolite may help in the differentiation of glial tumors on MV MRS.