Analysis of antimycin A by reversed-phase liquid chromatography/nuclear magnetic resonance spectrometry

A mixture of closely related streptomyces fermentation products, antimycin A, is separated, and the components are identified by using reversed-phase high-performance liquid chromatography with directly linked 400-MHz proton nuclear magnetic resonance detection. Analyses of mixtures of three amino acids, alanine, glycine, and valine, are used to determine optimal measurement conditions. Sensitivity increases of as much as a factor of 3 are achieved, at the expense of some loss in chromatographic resolution, by use of an 80-microL NMR cell, instead of a smaller 14-microL cell. Analysis of the antimycin A mixture, using the optimal analytical high-performance liquid chromatography/nuclear magnetic resonance conditions, reveals it to consist of at least 10 closely related components.