This study aimed to evaluate the properties of cellulose nanofibers (CNFs) with different hemicellulose contents and cellulose II polymorphs. A link was found between these polysaccharides and the properties of CNFs. A decrease in crystallinity (from 69 to 63%) and changes in the crystalline structure of cellulose subjected to an alkaline environment were observed, promoting the partial conversion of cellulose I to cellulose II (from 2 to 42%) and preventing CNFs production at NaOH concentrations higher than 5%. Most treatments showed pseudoplastic fluid behavior, except for the 10% NaOH treatment over 2 h, which showed Newtonian fluid behavior. The quality index of the reference CNFs (TEMPO-oxidized) was the highest (80 ± 3), followed by that of the 5% NaOH-treated (68 ± 3 and 22% energy savings compared to the untreated sample), and the untreated (63 ± 3) samples; and the 10% NaOH treatments had quality indices of 51 ± 3 and 32 ± 1, respectively.