A Practical Method for Preparing Copper-Free Bleomycin A2

  • Akio Fujii


A solution of 3.0 g of copper-containing bleomycin A2 hydrochloride (930 meg potency/mg. 4.28% copper content) in 100 ml of distilled water was passed through a column packed with 1 liter of Amberlite XAD-2 in distilled water to effect adsorption of bleomycin A2.Then 3 liters of a 5% aqueous solution of Na-EDTA was applied to the column to remove Cu-EDTA. The column was washed with 1 liter of a 5% sodium chloride solution and then with 2 liters of distilled water. Finally, 1.5 liters of a mixture (1: 1) of 2.5 mM hydrochloric acid and acetone was allowed to flow through the column to elute bleomycin A2.The latter half of the eluate, 700 ml in volume, was collected and concentrated under diminished pressure. The residue was subjected to reprecipitation from water-acetone, and dried to obtain 2.8 g of copper-free bleomycin A2 hydrochloride (945 meg potency/mg; 0.0083% copper content) in the form of a pale yellow powder; yield 93.3%, absorbance (aqueous solution) at 292 nm \( E_{1{\text{cm}}}^{1\% } = 103. \)


Public Health Aqueous Solution Analytical Chemistry Sodium Chloride Hydrochloric Acid 
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  1. 1.
    T. Takita, A. Fujii, Y. Muraoka, S. Mizuguchi, and H. Umezawa, U.S. Patent No.3, 929, 993 (1975).Google Scholar

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© Springer-Verlag New York Inc. 1979

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  • Akio Fujii

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