Purification of Chitin from Pupal Exuviae of the Black Soldier Fly

Abstract Purpose Chitin purification from remains (pupal exuviae after metamorphosis to adult flies) of Hermetia illucens farming was optimized performing demineralization, deproteinization and bleaching under different conditions. The optimal parameters to obtain high-purity chitin were determined. Methods Dried and ground pupal exuviae, whose composition was initially determined, were demineralized using six different acids. Proteins were removed with a NaOH treatment in which temperature, molarity and duration were varied in a randomized experiment. Bleaching was carried out testing ten different chemicals, including NaOCl, H 2 O 2 , solvent mixtures and enzymes. The efficiency of each step was determined to assess the optimal conditions for each of them. The resulting chitin was subjected to spectroscopic characterization. Results The highest demineralization efficiency (90%) was achieved using 0.5 M formic acid for 2 h at 40 °C, confirming the validity of organic acids as a more sustainable alternative to inorganic acids. The treatment with 1.25 M NaOH at 90 °C for 4 h showed the highest deproteinization efficiency, removing 96% of the proteins. Temperature and NaOH concentration were the significant parameters for deproteinization efficiency. The most efficient bleaching treatment was with 6% NaOCl at 60 °C for 1 h (67% efficiency). H 2 O 2 could also be a valid alternative to avoid environmental risk related to chlorine-containing compounds. At the end of the purification process 17% of the original biomass was retained with a chitin content of 85%, corresponding to a chitin yield of 14% related to the initial biomass. Solid-state nuclear magnetic resonance showed that the purified chitin had a degree of acetylation of 96% and X-ray powder diffraction gave a crystallinity index of 74%. Conclusion This investigation shows an optimized method for extraction of high-purity chitin from H. illucens pupal exuviae, supporting the validity of insect-farming remains as source of this versatile biopolymer. Graphical Abstract