Carboxymethyl chitosan (CM-chitosan) was prepared by chemical reaction with monochloroacetic acid under various conditions, and the chemical structure was analyzed by IR and NMR. The water solubility of the CM-chitosans had close relationships to the modifying conditions and the degree of carboxymethylation. The CM-chitosans, prepared at temperatures of 0-10 8C were soluble in water. But the CM-chitosan prepared between 20 and 60 8C were insoluble in the water at near-neutral pH. The water insolubility of CM-chitosans at various pHs varied with the degree of carboxymethylation. The increase in reaction temperature increased the fraction of carboxymethylation and increased the insolubility at lower pHs; the increase of the ratio of water/isopropanol in the reaction solvent decreased the fraction of carboxymethylation and increased the insolubility at higher pHs.

The protonation constants of chitosans with different molecular weight (Mw) and degree of deacetylation were demonstrated by the modified Henderson-Hasselbalch model and Hogfeldt’s three-parameter model. The protonation constants pKa of chitosan showed a slightly decreasing from 6.51 to 6.39 when the molecular weight changing from 1370 to 60 kDa. The degree of deacetylation showed a greatly effect on pKa values, which were increased from 6.17 to 6.51 with the degree of deacetylation decreasing from 94.6 to 73.3%. The degree of deacetylation influenced the balance of hydrophobic interactions and hydrogen bondings on chitsoan. The fitting of the titration data to both two methods showed that Hogfeldt’s three-parameter model provided a better agreement. The results obtained in this paper allowed to a better understanding of some physicochemical mechanisms and biological properties of chitosan.

In a new approach to the preparation of solid chitosan acetate, the dependence of solubility of chitsoan acetate on the mole ratio of acetic acid to GlcN residues of chitosan was evaluated from turbidity. The structure of the product chitosan acetate was characterized by titration and FT-IR. It was demonstrated that the chitosan acetate with high solubility retained the structure and antibacterial activity of chitosan. The antibacterial activities against Escherichia coli (Gram negative) and Staphylococcus aureus (Gram positive) have been exhibited by a special kind of agar and further investigated by optical density method. It was found that chitosan acetate at a concentration of 0.1% (m/V) exhibited some antibacterial activity but some of the bacteria did still grow. With a concentration of 0.15% (m/V), there were nearly no bacteria grown. Electron micrographs revealed bacterial action patterns of chitosan acetate towards E. coli and S. aureus.

Chitosan was modified by coupling with linoleic acid through the 1-ethyl-3-(3-dimethylaminopropyl)- carbodiimide-mediated reaction to increase its amphipathicity for improved emulsification. The micelle formation of linoleic acid-modified chitosan in the 0.1 M acetic acid solution was enhanced by O/W emulsification with methylene chloride, an oil phase. The fluorescence spectra indicate that without emulsification the self-aggregation of LA-chitosan occurred at the concentration of 1.0 g/L or above, and with emulsification, self-aggregation was greatly enhanced followed by a stable micelle formation at 2.0 g/L. The addition of 1 M sodium chloride promoted the self-aggregation of LA-chitosan molecules both with and without emulsification. The micelles of LA-chitosan formed nanosize particles ranging from 200 to 600 nm. The LA-chitosan nanoparticles encapsulated the lipid soluble model compound, retinal acetate, with 50% efficiency.

Membranes were prepared from chitosan with different molecular weights by a casting method. The molecular affinity and permeability of the membranes for sodium chloride, glucose, tyrosine, and bovine serum protein were measured at 4