Separation of spiramycin components using high-speed counter-current chromatography

High-speed counter-current chromatography was successfully applied to the separation of the main components of spiramycin. A 25-mg quantity of sample was separated using a two-phase solvent system composed of n-hexane–ethyl acetate–methanol–water (3:6:5:5), and the fractions were analyzed by high-performance liquid chromatography and fast atom bombardment mass spectrometry. The separation yielded 13.4, 0.7 and 1.7 mg of spiramycins I, II, and III with purities of 98.2, 92.3 and 97.4%, respectively.