Preparation and characterisation of chitosans with oligosaccharide branches Original Research Article

The trimer 2-acetamido-2-deoxy-d-glucopyranosyl-β-(1→4)-2-acetamido-2-deoxy-d-glucopyranosyl-β-(1→4)-2,5-anhydro-d-mannofuranose (A–A–M) was reductively N-alkylated onto a fully de-N-acetylated chitosan (FA<0.001, DPn=25) to obtain branched chitosans with degree of substitution (DS) of 0.070, 0.23 and 0.40, as determined by 1H NMR spectroscopy. The apparent pKa values of the primary and secondary amines of the chitosans substituted with the trimer A–A–M were determined by monitoring the chemical shift of the H-2 of GlcN, and were determined as 6.5–6.9 for the primary (unsubstituted) amines and as 5.0–5.2 for the secondary (substituted) amines. The intrinsic pKa values (pKint) were found to be 7.3–7.4 for the substituted and 8.7 for the unsubstituted amines. The chitosan branched with A–A–M (DS 0.40) was found to be soluble in aqueous solution over the entire pH range. SEC-MALLS (size-exclusion chromatography with a multi-angle laser light scattering detector) further showed that addition of branches did not affect the molar hydrodynamic volume of the chitosan.