Nanocellulose-based biomaterials for biomedical and pharmaceutical applications have been extensively explored. However, studies on different levels of impurities in the nanocellulose and their potential risks are lacking. This article is the most comprehensive to date survey of the importance and characterization of possible leachables and extractables in nanocellulose for biomedical use. In particular, the (1,3)-β-d-glucan interference in endotoxin detection in algal nanocellulose was addressed. Potential lipophilic and hydrophilic leachables, toxic heavy metals, and microbial contaminants are also monitored. As a model system, nanocellulose from Cladophora sp. algae is investigated. The leachable (1,3)-β-d-glucan and endotoxin, which possess strong immunogenic potential, from the cellulose were minimized to clinically insignificant levels of 4.7μg/g and 2.5EU/g, respectively. The levels of various impurities in the Cladophora cellulose are acceptable for future biomedical applications. The presented approach could be considered as a guideline for other types of nanocellulose.