Lissahatina Fulica shell waste contains chitosan with the potential for antioxidants and hypercholesterolemia. This study, therefore, reports on the analysis of molecular characterization, antioxidant assay, and acute lethal dose 50 (LD50) toxicity in Lissahatina Fulica shell waste chitosan rats using the hot plate magnetic stirrer technique. Chitosan was performed to analyze water content, protein, functional groups, molecular weight, particle size, morphology, antioxidants, and acute toxicity test. The researchers found that the hot plate magnetic stirrer treatment affected the water content, molecular weight, particle size, morphology, antioxidant, and non-toxicity. In Lissahatina Fulica shell waste chitosan, the water content was 0.345%, the protein content was 1.87%, the molecular weight was 2.297 kDa, the deacetylation degree value was 92.37%, and the particle size was 59 µm. Surface roughness had an Rp value of 249, Rv value of 0, Ra value of 81.5, Rq value of 97.8, and IC50 value of 2.35 µg/mL. In addition, the chitosan dose of 3000mg/kg BW did not cause death in the sample rats. In conclusion, chitosan from Lissahatina Fulica shell waste has the potential to be developed in the pharmaceutical industry for further development.