Asymmetry in MT causes erroneous GluCEST computation. The study focuses on removal of MT effects from Z-spectra for better quantification of GluCEST contrast using in-vivo human volunteer at 7T and rat brain with tumor at 9.4T. Different lineshapes for modelling and fitting broad MT spectrum has been compared. Lorentzian lineshape provided significant difference between GluCEST contrast before and after MT removal and also preserves the gray-matter to white-matter glutamate concentration ratio validated with MR spectroscopic based estimation. After removal of MT effect using Lorentzian lineshape, the specificity of GluCEST to glutamate concentration increases which can helps better diagnosis of diseases.