Determination of the pattern of acetylation of chitosan samples: Comparison of evaluation methods and some validation parameters

Chitosan is a linear copolymer consisting of glucosamine and N-acetylglucosamine units. Its specific biological features make it a very attractive biomaterial for diverse applications in both pharmaceutics and medicine. The pattern of acetylation of chitosan samples (P_A) can strongly influence their biomedical activities. So far, three methods of determining the P_A value of chitosan samples, based on NMR spectroscopy, have been developed. Chitosans with a degree of acetylation (F_A) from 0.02 to 0.48 were used to compare these methods. The most suitable and specific method of P_A determination was ¹³C NMR based on the intensities of carbon C-5 signals. Some validation parameters—specificity, sensitivity, precision (repeatability and reproducibility)—were established. The intra- and inter-day precision of this method depended on the degree of acetylation of the chitosan samples. It was found to be specific, sensitive, repeatable and reproducible.