Purpose CEST MRI experiments of mobile macromolecules, for example, proteins, carbohydrates, and phospholipids, often show signals due to saturation transfer from aliphatic protons to water. Currently, the mechanism of this nuclear Overhauser effect (NOE)‐based transfer pathway is not completely understood and could be due either to NOEs directly to bound water or NOEs relayed intramolecularly via exchangeable protons. We used glycogen as a model system to investigate this saturation transfer pathway in sugar polymer solution. Methods To determine whether proton exchange affected saturation transfer, saturation spectra (Z‐spectra) were measured for glycogen solutions of different pH, D2O/H2O ratio, and glycogen particle size. A theoretical model was derived to analytically describe the NOE‐based signals in these spectra. Numerical simulations were performed to verify this theory, which was further tested by fitting experimental data for different exchange regimes. Results Signal intensities of aliphatic NOEs in Z‐spectra of glycogen in D2O solution were influenced by hydroxyl proton exchange rates, whereas those in H2O were not. This indicates that the primary transfer pathway is an exchange‐relayed NOE from these aliphatic protons to neighboring hydroxyl protons, followed by the exchange to water protons. Experimental data for glycogen solutions in D2O and H2O could be analyzed successfully using an analytical theory derived for such relayed NOE transfer, which was further validated using numerical simulations with the Bloch equations. Conclusion The predominant mechanism underlying aliphatic signals in Z‐spectra of mobile carbohydrate polymers is intramolecular relayed NOE transfer followed by proton exchange.