Oxidized cellulose (OC) nanofiber was successfully prepared from the dry sheet of Nata De Coco (DNDC) using the mixture system of HNO 3 /H 3 PO 4 -NaNO 2 for the first time. e carboxyl content of the OC was investigated at different conditions (HNO 3 /H 3 PO 4 ratios, reaction times, and reaction temperatures). e results revealed that the carboxyl content of the OC increased along with the reaction time, which yielded 0.6, 14.8, 17.5, 20.9, 21.0, and 21.0% after 0, 6, 12, 36, and 48 hours, respectively. e reaction yields of the OC ranged between 79% and 85% when using HNO 3 /H 3 PO 4 ratio of 1 : 3, 1.4% wt of NaNO 2 at 30°C at different reaction times. From the structural analysis, the OC products showed a nanofibrous structure with a diameter of about 58.3-65.4 nm. e Fourier transform infrared spectra suggested the formation of carboxyl groups in the OC after oxidation reaction. e crystallinity and crystalline index decreased with an increase of reaction time. e decrease of crystallinity from oxidation process agreed with the decrease of degree of polymerization from the hydrolysis of β-1,4-glycosidic linkages in the cellulose structure. e thermal gravimetric analysis results revealed that the OC products were less thermally stable than the raw material of DNDC. In addition, the OC products showed blood agglutinating property by dropping blood on the sample along with excellent antibacterial activity.