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Acetylsprimycin (ASPM) is known to be a kind of potent macrolide antibiotics in vivo.It has four active components(mono-ASPM Ⅱ, Ⅲ and bi-ASPM Ⅱ, Ⅲ) coexisting and their total contents should be no less than 75% in the tablet according to Chinese pharmacopoeia.For determination of the total contents of the four ASPM active components, an HPLC method, in which using C18 colunmn, acetonitrile-0.1 mol L-1 NH4Ac(60∶40, pH 7.2) eluent, and UV detector 232nm, was adopted in the Chinese pharmacopoeia.