A series of propyl cellulose (PC) samples with different degrees of substitution (DS) ranging from 0.34 to 2.02 were prepared by a slurry method using propyl bromide as the etherification reagent. Two-dimensional nuclear magnetic resonance (NMR) studies were performed to identify the 1H and 13C chemical shifts of eight anhydroglucose units (AGUs) in PC chains including un-, 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, 3,6-di-, and 2,3,6-tri-substituted ones. In addition, the mole fractions (χ) of these AGUs in the studied PC samples and their changes with DS were determined from the quantitative 13C NMR spectra. The obtained χ−DS profiles were different from those of methyl and ethyl celluloses prepared by a similar slurry method, indicating that the molecular sizes of the substituent reagents utilized for cellulose ethers strongly affected their substituent distributions.