Separation of nucleotides by high-voltage capillary electrophoresis.

Abstract

A new separation method using high-voltage capillary electrophoresis was applied to the analysis of several nucleotides. This system consisted of a micro-bore glass capillary column of 80-micron i.d., a high-voltage power supply, and a uv detector set at 254 nm. Seven nucleotides (cyclic AMP, AMP, ADP, ATP, GMP, GDP, and GTP) were separated completely from each other in 0.02 M phosphate buffer (pH 7) containing 0.5% ethylene glycol by applying about 150 V/cm. The theoretical plate number for AMP was nearly 45,000. A good separation of nucleotides in biological samples of rat blood, liver, and kidney was attained under the same conditions as above. Concentrations of nucleotides in these biological samples were measured by using pyridoxamine as an internal standard.