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In brief, chitosan (0.5 g) was dissolved in pH 5.5 deionized and distilled water (DDW, 50 mL). Hydrocaffeic acid (591.0 mg) and EDC (1244.9 mg) in 25 mL of DDW and ethanol (1:1 v/v) were added to the chitosan solution, and reacted for 12 h. The pH of the reaction solution was maintained at 5.5 by the addition of 1 M HCl. After ethanol was evaporated, the product was purified by dialysis (MWCO: 12 00014 000, SpectraPor) against pH 5.0 HCl solution inDDWfor 2 days andDDWfor 4 h. The final product was freeze-dried and kept in a moisture-free desiccator before use.
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