The chitosan derivatives bearing different 4-chlorophenylcarbamate urea structures were synthesized under different reaction conditions, and their chiral recognition abilities were evaluated as chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC). These CSPs coated on silica gel exhibited different recognition ability depending on their structures and could be used with a few prohibited solvents as the eluent, which efficiently improved the resolution for some racemates.