The nucleation of hydroxyapatite (HAp) crystal through chemical interaction with collagen was investigated. A collagen membrane was soaked in a supersaturated simulated body fluid (1.5 SBF) solution with ion concentrations at 1.5 times that of normal simulated body fluid (1.0 SBF). A few carbonate-containing HAp crystals were formed mostly on the edge-side of the collagen membrane after 4 weeks, In the Fourier-transform infrared spectometry (FTIR) results, the carboxylate band of the collagen membrane showed red chemical shifts after the formation of HAp crystals, which coincided well with the decrease of the calculated bond orders of the carboxylate group when chelated with a calcium ion, which emulated the first-step nucleation of HAp crystal on the carboxylate group of collagen. The result implies that the binding of a calcium ion to the carboxylate group of collagen is one of the key factors for the nucleation of HAp crystals in a 1.5 SBF solution.