High-pressure liquid chromatography of free nucleotide patterns in normal and abnormal keratinocytes

A method for analyzing free nucleotides in the epidermis of the guinea pig is presented. Free nucleotides were extracted by using a methylalethanol mixture, and the analysis was carried out by high-pressure liquid chromatography on a column of Lichrosorb-NH2 with a single buffer of potassium phosphate. The concentration of total free nucleotides in the epidermis is about 4 times greater than that in the liver, kidney, spleen, or intestinal epithelium.l The free nucleotide level is markedly elevated in the hyperkeratotic epidermis induced by n-hexadecane. The alternation of free nucleotides in hyperkeratotic epidermis is discussed in relation to nucleic acid content, DNase, disc-electrophoretic properties of DNase, and salvage pathway enzymatic activity. Significant increases in the enzyme activity of the salvage pathway and in neutral DNase were observed in the hyperkeratotic stage. However, the DNA content and acid DNase activity were decreased. It is suggested that the pool size of free nucleotides in the epidermis is affected by the salvage enzyme system.