The objective of this work was to obtain Spirulina (Arthrospira maxima) nanoparticles (SNPs) by using high-impact mechanical milling and to characterize them by electron microscopy and spectroscopy techniques. The milling products were analyzed after various processing times (1–4 h), and particle size distribution and number mean size (NMS) were determined by analysis of high-resolution scanning electron microscopic images. The smallest particles are synthesized after 3 h of milling, had an NMS of 55.6±3.6 nm, with 95% of the particles being smaller than 100 nm. High-resolution transmission electron microscopy showed lattice spacing of ~0.27±0.015 nm for SNPs. The corresponding chemical composition was obtained by energy-dispersive X-ray spectroscopy, and showed the presence of Ca, Fe, K, Mg, Na, and Zn. The powder flow properties showed that the powder density was higher when the average nanoparticle size is smaller. They showed free flowability and an increase in their specific surface area (6.89±0.23 m2/g) up to 12–14 times larger than the original material (0.45±0.02 m2/g). Fourier transform infrared spectroscopy suggested that chemical damage related to the milling is not significant.