The invention relates to calcitonin long-acting slow-release microspheres which comprise the following components in percentage by weight: 0.5%-4% of acidic glucosan, 1.5%-12% of calcitonin and 84%-98% of slow-release or controlled-release high molecules. The particle diameter of each calcitonin long-acting slow-release microsphere is 1-200 micrometers. A plurality of calcitonin long-acting slow-release microspheres which contain different slow-release or controlled-release high molecules are combined and different calcitonin long-acting slow-release microspheres which are mutually separated and mutually continued in the concentrated release time are selected for combination to obtain the calcitonin long-acting slow-release microsphere combination. The invention also discloses a preparation method of the calcitonin long-acting slow-release microspheres. The preparation method disclosed by the invention has the advantage that the slow-release or controlled-release high molecules can be completely released by reducing the adsorption of the calcitonin on the slow-release or controlled-release high molecules by causing a local acid environment by adding the acidic glucosan, thereby overcoming the defect that a protein polypeptide medicament is easy to be adsorbed in a slow-release material without being released; and the calcitonin long-acting slow-release microsphere combination which has the advantages of stability, long action, slow release and less burst release through the combination of the calcitonin microspheres in different prescriptions can be obtained.