This study is focused on the ability of apatite formation on the surface of nano-hydroxyapatite (HA)/ chitosan (CH) composite in simulated body fluid (SBF) in vitro. At first, natural nano-HA was prepared according to a wet-balling method and the composite was prepared by combining the natural nano-hydroxyapatite and chitosan, and then in vitro biomineralization test of natural nano-HA/CH composite was carried out in standard SBF. Subsequently, the quantity of the weight of the particles formed on the composite surface in SBF was measured by analytical balance, and the morphology change on the surface of the composite was observed by a scanning electron microscope (SEM). Lastly, a Fourier transform infrared spectroscope (FTIR) was used to investigate the chemical components of the particles formed on the natural nano-HA/CH composite surface in SBF. The result of quantity assessment shows that the weight of the composite increased with the increase of soaking time. The SEM image shows that the particles were gradually formed on natural nano-HA/CH composite surface, and the FTIR spectrum of the particles on composite surface confirms that these particles were carbonate apatite. This study indicates that the nano-HA/CH composite has a good ability for apatite formation in SBF, which predicts the bone-inducing ability of natural nano-HA/CH composite in vivo.