The chromatographic method for the determination of the intermolecular acetyl distribution of cellulose triacetate (CTA) was established; the key point of the method is a gradient elution by gradually changing the solvent composition of the ternary mixture of chloroform, methanol, and water on a phenyl-bonded silica gel stationary phase. A positive correlation between the retention time and the degree of acetyl substitution was observed under the established chromatographic conditions. The validity of the elution peak width as the measure of intermolecular acetyl distribution was confirmed by the co-injection and fractionation experiments and the comparison of a peak width with a statistically estimated distribution.