Disclosed are crystalline levoisovalerylspiramycin II and corresponding preparations, preparing methods and uses thereof. The preparations comprise crystalline levoisovalerylspiramycin II and pharmaceutically acceptable carrier and/or adjuvant, wherein the purity of levoisovalerylspiramycin II is above 90 wt%. The crystalline levoisovalerylspiramycin II has a good antibacterial activity, and the preparations include solution for injection, powder for injection or lyophilized powder for injection.