Die Reingewinnung des Digitonins

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Abstract
Com-mercial (Merck) digitonin was purified by paper chromatography and counter current distribution. The commercial product was 1st recrystallized 4 times from 85% ethanol, and then subjected to chromatography using as the solvent the upper phase of a mixture of butanol-water-glacial acetic acid, 4:5:1. Two bands were obtained, Rf 0.36 and 0.51 (substances I and II). The bands were eluted, and subjected to a 24-tube counter current distribution analysis. Substance I was homogeneous; II gave 2 peaks which could be separated. Biological assays of digitonin were carried out by causing it to be bound by cholesterol on the surface of red blood cells. Substance I had a cholesterol titer of 97.5; calculated for pure digitonin of M.W. 1229 would have been 100. II had a titer of 85; in this case however it was subsequently found that the M.W. was less than 1229. A sugar analysis revealed that I contained 2 molecules of glucose and galactose, and one of pentose; II contained one glucose molecule less. The infra-red spectrum of I, which is assumed to be pure digitonin, is given. Attempts to characterize the genins of the 2 substances were unsuccessful. It was also established that under the experimental conditions used the formation of the digitonide with cholesterol was independent of the magnitude of the digitonin excess.

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