The demand for hydroxyapatite (HA) is increasing annually due to the growing number of orthopedic patients, but it is a challenge to meet this growing demand without interfering with the integrity of the environment. Here, natural HA was produced from cow bone through ultrasound treatment followed by a calcination process at various temperatures. Characterization of the produced HA was performed by techniques such as thermogravimetric analysis, field emission scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction analysis. The elemental composition of the HA was quantified through X-ray photoelectron spectroscopy, energy dispersive X-ray spectroscopy, X-ray fluorescence, as well as carbon, hydrogen, nitrogen, sulfur (CHNS) analysis. The results demonstrate that pure HA was produced after calcination at 950°C, corresponding to pure HA phase.