Sea cucumber has a thick layer of skin consisting of lime components. In fact, the components contain chitin and chitosan, which have been recognized as potential sources materials for dietary supplement. This study aimed at evaluating the physical and chemical characteristics of chitosan nanoparticles extracted from sea cucumber Holothuria scabra when used as source materials for glucosamine. Chitin were extracted from dried samples, chitosan from chitin, while chitosan nanoparticles were obtained from chitosan with different concentrations (C₁ = 0.1%; C₂ = 0.2%; C₃ = 0.3%) of added sodium tripolyphosphate (NaTPP). Production process in this study resulted in 59.82% of chitosan extracted from chitin. Besides, the amount of chitosan nanoparticles obtained at 0.1%, 0.2%, and 0.3% additions of NaTPP were 90.6%, 92.8%, and 96.4%, respectively. These results were characterized in terms of whiteness degree (85.82%, 87.29%, 88.34%, respectively), deacetylation degree (90.6%, 95.8%, 96.2%), moisture (5.73%, 5.26%, 4.82%), and ash (1.29%, 1.07%, 0.98%). Looking at SEM and PSA tests, chitosan was morphologically found to be heterogeneously distributed with averaged 177-micron particle sizes. They also had larger particle chunks and solid as well as intact forms. Meanwhile, chitosan nanoparticles had smaller and smoother chunks, while they were produced in solid and intact forms. Besides, they were homogeneously distributed with sizes ranging between 134 − 206 nm (C₁), 114-128 nm (C₂), and 97-108 nm (C₃). Then, increments in NaTPP concentrations were discovered to contribute to the reduction of H. scabra-sourced chitosan nanoparticles size.