Objective To know the efficacy of rinse with fresh distilled water and stored distilled water in eliminating pyrogen.Methods Limulus amebocyte lysate test was used to examine the glass syringe which had been immersed in triple-effect pyrogen inactivator for washing off pyrogen.Results The pyrogen examination eligible rate of the syringe washed with fresh filtrated flowing distilled water was 98.3%,while that of the syringe washed with the distilled water which had been stored for 24 h was 65.0% and that of the syringe washed with the distilled water which had been stored for 48 h was 45.0%.Conclusion The glass appliances after treated for eliminating pyrogen should be washed with fresh flowing distilled water,while washing with distilled water which had been stored for longer than 24 h would influence the final efficacy in eliminating pyrogen.