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A method for chromatographic separation and quantitative determination of individual components of the antibiotic virginiamycin, produced by microbiological synthesis (Streptomyces virginiae strain 147), is described. The components, M1–2and S1–5, were isolated from fermentation broth and identified by HPTLC and HPLC (the results obtained using the two methods correlate well with each other). The conditions of culturing of the producer and the composition of nutritive media were optimized. Using UV irradiation as a mutagenic factor, the producer was selected for increased level of synthesis of the antibiotic; this was achieved by inducing mutations that impart resistance to virginiamycin and meta-fluorophenylalanine, an analog of phenylalanine.