Preparation, Characterization, and Biochemical Activities of N-(2-Carboxyethyl)chitosan from Squid Pens

Chitosan was prepared by alkaline N-deacetylation of β-chitin from squid pens, and N-carboxyethylated derivatives (N-CECS) with different degrees of substitution (DS) were synthesized. The carboxyethylation of the polysaccharide was identified by Fourier transform infrared, 1H and 13C nuclear magnetic resonance (NMR), and X-ray diffraction analyses. The DS of the derivatives was calculated by 1H NMR and elemental analysis. All three N-CECS samples showed good water solubility at pH > 6.5. The antioxidant properties and bile acid binding capacity of the derivatives were studied in vitro. The highest bile acid binding capacity of all N-CECS reached 36.9 mg/g, which was 2.6-fold higher than that of chitosan. N-CECS showed a stronger scavenging effect on 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical ability, and EC50 values were below 2 mg/mL. The scavenging ability of N-CECS against superoxide radicals correlated well with the DS and concentration of N-CECS. These results indicated that N-carboxyethylation is a possible approach to prepare chitosan derivatives with desirable in vitro biochemical properties.