Revised Methods for Purification of Ribosyl-trans-zeatin from Vinca rosea L. Crown Gall Tumor Tissue

Ribosyl-trans-zeatin has been purified from Vinca rosea L. crown gall tumor tissue by using two new sequences of isolation procedures. Identification of the compound has been established by mass spectrometry, ultraviolet absorbancy spectra, chromatographic values, and growth activity. The isolation sequences eliminate the exposures to pH extremes and the strong cation-exchange resin used in the purification reported earlier. The initial extraction procedures have been designed so as to avoid enzymatic alteration or production of active material and to prevent the inclusion in the extracts of nucleic acids which might serve as soures of the small active compounds. The production of ribosylzeatin by the tumor tissue is confirmed and the validity of isolation steps such as the use of cation exchangers is supported.