The relevance of equilibrium in a multifragmentation reaction of very central 40Ca+40Ca collisions at 35 MeV/nucleon is investigated by using simulations of Antisymmetrized Molecular Dynamics (AMD). Two types of ensembles are compared. One is the reaction ensemble of the states at each reaction time t in collision events simulated by AMD, and the other is the equilibrium ensemble prepared by solving the AMD equation of motion for a many-nucleon system confined in a container for a long time. The comparison of the ensembles is performed for the fragment charge distribution and the excitation energies. Our calculations show that there exists an equilibrium ensemble which well reproduces the reaction ensemble at each reaction time t for the investigated period 80 ≤ t ≤ 300 fm/c.