Disclosed are levoisovalerylspiramycin I, II or III, preparations, preparingmethods anduses thereof. The preparations comprise levoisovalerylspiramycin I, II or IIIandpharmaceutically acceptable carrier and/or adjuvant, wherein the purity oflevoisovalerylspiramycin I, II or III is above 90 wt%. Thelevoisovalerylspiramycin I, II orIII has a good antibacterial activity, and the preparations include solutionfor injection,powder for injection or lyophilized powder for injection.