Dynamic glucose enhanced(DGE) MRI has shown promise in glucose metabolism studies. In recent studies, a hypertonic dextrose solution was used for reliable detection of glucose in the brain. However, the effects of the hypertonic solution on DGE signal have not been verified yet. This study aimed to investigate the signal contributions from non-glucose related components. We used hypertonic D-, L-glucose, and NaCl solution to identify osmolality effects. Our data show an osmotic shift of water between the extravascular and intravascular space, induced by administering D-glucose(50%), can highly affect the DGE signal but negligible contributions were observed from the intravascular space.