Three simple, precise, and sensitive UV spectrophotometric methodshave been developed and validated for the determination of Chitosan (CH) in the presence of Ascorbic acid (AA) in a binary mixture. Method A is a derivative ratio method (¹DD) that measures the peak amplitude of the first derivative for CH and AA at 207.2 and 282 nm; respectively. Method B is a ratio difference method (RDSM) that measures the peak amplitude difference of the ratio spectra (DP_{204.6- 202.9}) and (DP_268-230) for CH and AA; respectively. Method C is a mean centering of ratio spectra method (MCR) that measures CH and AA at 204.6 and 269.1 nm; respectively. Method A, B, and C were able to successfully determine the studied drugs in a concentration range of 0.5-9 mg/mL for CH and 2-20 mg/mL for AA. All results were statistically compared with the reported methods, where no significant difference was observed. The proposed methods were applied to the analysis of the studied drugs in pure and pharmaceutical formulations.