Thin-layer chromatography of nucleotides on DEAE- and ECTEOLA-cellulose is superior to the methods of seperation in use up to the present. Using dilute hydrochloric acid, and without a gradient elution technique, it is possinle to acheive a group seperation of monodi-, and tri-phosphates as well as subfractionation within these groups depending on the basicity of the nucleobases. Thin-layer chromatography is appreciably more sensitive than paper chromatography; 5×10−4 mole adenine neuclotides can be still be detected. The total quantity of material that can be chromatographed without tailing is greater in this procedure than that with paper chromatography; a mixture consisting of 1 μmole ATP and ADP per starting spot can be resolved with ease . Hence the procedure can also be applied for micro-scale preparative seperations . A mixtures of 10−2 μmole ADP and ATP, respectively, can be seperated completely in four to ten minutes.