A Walkway from Crayfish to Oligochitosan

Edible crayfish are an object of local fishing and artificial breeding in many countries. This industry is very promising in terms of production of healthy foods and byproducts, such as biologically active polyaminosaccharide-chitosan and its derivatives. However, crayfishing is far from being at the level at which it could be. This laboratory scale protocol describes a walkway from crayfish Actacus leptodactylus to chitin, chitosan, and oligochitosan hydrochloride, with the main emphasis on the way of getting rid of the impurities (residual heavy metals, proteins and other residues) commonly present in commercial chitosan and its derivatives, as well as the characterization of the products by means of inductively-coupled plasma mass spectrometry (ICP-MS), energy-dispersive X-ray spectroscopy (EDXS), protein and elemental analysis, proton magnetic resonance spectroscopy (H-1 NMR), and chromatography methods. The protocol includes the preparation of crude shell waste; the extraction of proteins from crude shell waste and preparation of deproteinated shell waste, demineralization and decolorization of the deproteinated crayfish shell waste, deacetylation of chitin, and depolymerization of chitosan. EDXS shows the presence of Al and Si residues in chitin is found when the deproteination of crayfish waste is carried out in an alumosilicate glass vessel. In contrast, these residues are absent when deproteination is carried out in the borosilicate glass flask. Analytical data show that the content of residues in chitosan and oligochitosan hydrochloride meets pharmaceutical requirements. The study demonstrates crayfish waste a promising for the purification of chitosan, for the preparation of pharmaceutical grade oligochitosan hydrochloride, and can improve commercialization of crayfishes.