In-vivo Safety Evaluation of Chitosan Extracted from the Pupal Exuviae of Hippotion celerio L.

Abstract

Chitosan, an important and abundant natural-based polymer, was extracted for the first time from the pupal exuviae (PE) of Hippotion celerioL., reared and fed with an invasive plant species, Colocasia esculenta (L.) Schott. In this study, we investigatedthe safety and biotechnological potential of insect-based chitosan. Demineralization and deproteinization were performed with 1 M hydrochloric acid at 25°C for 3 hours at 300 RPM and 1 M sodium hydroxide (NaOH) for 4 hours at 100°C, respectively. The extracted chitin was converted into chitosan via deacetylation using 50% NaOH for 4 hours at 80°C. Both chitin and chitosan were characterized by Fourier Transform Infrared Spectroscopy (FTIR) to determine its purity and Scanning Electron Microscopy (SEM) to examine its surface morphology. For the acute toxicity study, a single dose of three concentrations of 300, 2000, and 5000 mg/kg bw of chitosan were administered orally using a gavage to healthy female Swiss albino mice. The chitin and chitosan yield of the dry weight were 21.74±2.15% and 58.26±4.24%, respectively. Additionally, the degree of deacetylation (DD) was found to be 85%, higher compared to the commercial chitosan obtained from the shrimp with 83%. Scanning Electron Microscope (SEM) exposed smooth, porous, and fibrous surfaces, suggesting its use in biotechnology. LD50value for chitosan in Swiss albino mice was greater than 5000 mg/kg bw. Thus, this study suggests that chitosan extracted from the pupal exuviae of H. celerio L. is safe and could beused in future applications in tissue engineering, biomedicine, and textile industry.