Cellulose nanocrystals (CNCs) were successfully prepared from Sansevieria trifasciata fibers (STFs) via ammonium persulfate (APS) oxidation in this study. The influences of the APS concentration (1.1, 1.5, and 1.9 M) and oxidation temperature (60, 70, and 80 °C) on the characteristics of CNCs were studied. The resulting CNCs were characterized using Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and thermogravimetric analysis (TGA). The TEM observations revealed that the rod-like CNCs possessed average length and diameter ranges of 96 to 211 nm and 5 to 13 nm, respectively, which led to an aspect ratio range of 16–19. The optimum conditions for maximum crystallinity were achieved at an oxidation temperature of 70 °C, a reaction time of 16 h, and an APS concentration of 1.5 M. All CNCs exhibited lower thermal stability compared to the STFs. The CNCs could be produced from the STFs through the APS oxidation process and showed potential as nanocomposite reinforcement materials.