In this study, a method for the characterization and quantification of chitosan was designed using acid
hydrolysis, glucosamine derivatization, and high performance liquid chromatography. Based on a kinetic
study of acid hydrolysis, we have demonstrated that chitosan can be quantitatively hydrolyzed into
glucosamine in 6 h with either 10 M hydrochloric acid (HCl) at 105 ◦C, or 12 M HCl at 90 ◦C. Following N-(9-
fluorenylmethoxycarbonyloxy) succinimide (Fmoc-OSu) derivatization, the glucosamine content can be
separated from the rest of the hydrolysates and quantified using reverse-phase HPLC with UV detection.
This method was validated for linearity, precision (repeatability and reproducibility), and accuracy using
both chitosan and a dietary supplement formulation containing chitosan.