Colchicine, labeled with tritium in the acetate group, has been prepared with a specific activity of 250 mc/mmole. U.S.P. colchicine was purified, then hydrolyzed to trimethylcolchicinic acid. The trimethylcolchicinic acid was next methylated to a mixture of deacetylcolchicine and isodeacetylcolchicine. The deacetylcolchicine was separated from the isodeacetylcolchicine by silica gel column chromatography and acetylated to colchicine with tritiated acetic anhydride. The newly synthesized colchicine (acetyl-3H) was separated from the reaction mixture and purified on a Bio-Sil-A silicic acid column. A method for identification of colchicine
"1CO2 produced in the Brookhaven 152-cm cyclotron was converted to formaldehyde, which in turn was used for the enzymatic conversion of deoxyuridine-5'-phosphate to ["Cithymidylate. Enzymatic treatment of the nucleotide with alkaline phosphatase gave ["C]thymidine.
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