DISCLOSED ARE LEVOISOVALERYLSPIRAMYCIN I, II OR III, PREPARATIONS, PREPARING METHODS AND USES THEREOF. THE PREPARATIONS COMPRISE LEVOISOVALERYLSPIRAMYCIN I, II OR III AND PHARMACEUTICALLY ACCEPTABLE CARRIER AND/OR ADJUVANT, WHEREIN THE PURITY OF LEVOISOVALERYLSPIRAMYCIN I, II OR III IS ABOVE 90 WT%. THE LEVOISOVALERYLSPIRAMYCIN I, II OR III HAS A GOOD ANTIBACTERIAL ACTIVITY, AND THE PREPARATIONS INCLUDE SOLUTION FOR INJECTION, POWDER FOR INJECTION OR LYOPHILIZED POWDER FOR INJECTION. MOST ILLUSTRATIVE DIAGRAM: FIGURE 1