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We have reported the isolation') and the singlecrystal X-ray analysis' of a novel neutral macrolide antibiotic cytovaricin, from the culture of a Streptomyces sp. In this paper we report the isolation of methyl-;3-D-cymaroside as a methanolysis product of cytovaricin, which confirms the absolute configuration of cytovaricin as that shown in Fig. 1. Cytovaricin (200 mg) was dissolved in 0.4 ml of 0.25 % methanolic hydrogen chloride and left overnight at room temperature. Four products (one major and three minor) were revealed by spraying the TLC chromatogram of the methanolysis product with anisaldehyde-sulfuric acid. The reaction mixture was neutralized with dilute ammonium hydroxide and concentrated in vacuo to dryness. The residue was chromatographed on a silica gel column with the solvent system, ethyl acetate benzene (1: 1). The fraction containing the major product was further purified by preparative HPLC [Nucleosil 5C9, methanol water (23: 77)]. The pure product (12 mg) was obtained by distribution between ether and saturated aqueous sodium chloride. 1H NMR data (400 MHz) with assignment are

The absolute configuration of cytovaricin: isolation of methyl .BETA.-D-cymaroside by methanolysis.