Abstract
In previous reports 1 2 we have shown that extracts of the urine of patients with leucemia contained substances capable of inciting cellular proliferation in the organs of guinea pigs similar to those found in human leucemias. In one of these reports 2 it was shown that the substances obtained were specific for the type of leucemia from which the urinary extracts were made.
It has seemed obvious that the chloroform extracts described in the earlier communication contained many inert materials as well as the active principles. Because of this we decided to fractionate the chloroform extracts and tried to trace the active principles preparatory to chemical analysis.
The following method has been developed to fractionate the extracts of urine of patients with chronic myeloid and chronic lymphoid leucemia. It applies particularly to the fractionation of the former.
The urine is extracted with chloroform as before. 2 The chloroform is removed by distillation and the residue is suspended in 200 to 300 cc of 10% sodium hydroxide. This suspension is extracted 8 to 10 times with 100 cc portions of ethyl ether to remove any substances which will not react with sodium hydroxide. The fraction soluble in ethyl ether (A) is recovered by removal of the solvent. The residual alkaline solution (B) is reacidified with 30 to 50 cc of concentrated hydrochloric acid. The acid solution is extracted with chloroform, the extract after removal of the chloroform is extracted with successive portions of petroleum ether (C); these extracts are combined, distilled in vacuo. The residue crystallizes readily in the cold.
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