AbstractThe mass‐specific charge distribution in molar mass fractions of cationic starch derivatives was investigated. The molar mass fractions were produced by semi‐preparative SEC (SP‐SEC). The derivatives were cut into an amylopectin‐rich fraction F1, an intermediate fraction F2, containing low molar mass amylopectin and high molar mass amylose, and an amylose‐rich fraction F3. The weight‐average molar mass (Mw) and molar mass distribution (MMD) of the fractions were determined by SEC with multi‐angle laser light scattering (SEC‐MALLS). The mass‐specific charge of each fraction was calculated from the consumption of anionic titrant solution using polyelectrolyte titration (PET) in combination with particle charge detection (PCD). The difference in substituent distribution between the fractions was tested by the Student's t‐test. The weight‐average molar mass of the starch derivatives was not dependent on the degree of substitution (DS) or the derivatization process. Depending on the DS value or derivatization process, different substituent distributions were observed. The results for the mass‐specific charge distribution in different molar mass fractions of cationic starch derivatives with graded DS between 0.015 and 0.130 from the slurry process were discussed. The heterogeneity of substituent distribution decreased with increasing DS of the starch derivative. This was the case for samples from both the slurry and semi‐dry processes. The heterogeneity of derivatization was highest for low DS samples up to DS 0.03, with the amylopectin‐rich fraction incorporating more charges than the amylose‐rich fraction. This was more pronounced for the sample from the slurry process than from the semi‐dry process.