The nano crystalline hydroxyapatite with highly uniform spherical morphology has successfully been prepared using the new cationic surfactant Cetrimide as template by co-precipitation method at ambient temperature. The sample was calcinated at 750 oC for 8h. The FTIR spectrum shows the chemical composition of HAP. The XRD pattern confirms the characteristic peaks and the hexagonal structure of HAP. The lattice parameters calculated from the XRD pattern are a = b ~ 0.93873 nm, c = 0.68486 nm, the axial ratio ~0.7296, the volume of the unit cell ~522.6535x10-30 m3. The size and morphology of the sample were analyzed by FESEM, which shows that the spherical HAp particles with diameter 150-200 nm have successfully been synthesized. All the inspections confirm the successful preparation of nano spherical HAP with excellent morphology control, and uniform size.