High Klason lignin content (23 wt%) cellulose nanofibrils (LCNF) were successfully isolated from eucalyptus pulp through catalyzed chemical oxidation, followed by high-pressure homogenization. LCNFs had a diameter of ca. 13 nm according to AFM evaluation. Dense films were obtained through vacuum filtration (nanopaper) and subjected to different drying methods. When drying under heat and mild vacuum (93 °C, 95 kPa) a higher water contact angle, lower roughness and oxygen transmission rate were observed, compared to those drying at room temperature under compression conditions. DSC experiments showed difference in signals associated to T_g of LCNF compared to CNF produced from spruce bleached pulp through enzymatic pre-treatment. The LCNF-based nanopaper showed mechanical properties slightly lower than for those made from cellulose nanofibrils, yet with increased hydrophobicity. In summary, the high-lignin content cellulose nanofibrils proved to be a suitable material for the production of low oxygen permeability nanopaper, with chemical composition close to native wood.