Separation of nucleotides by micellar electrokinetic chromatography

A novel method for the separation of nucleotides by micellar electrokinetic chromatography with diode array detection was developed. The effect of some key factors, such as concentration of buffer and sodium dodecyl sulfate (SDS), pH, surfactant, addition of organic solvent, capillary temperature and applied voltage on the separation of nucleotides was investigated. Under the optimized conditions, the baseline separation of five nucleotides was achieved with satisfactory repeatability and high separation efficiency in 6 min using 36 mmol/L borate buffer (pH 9.0) containing 3% (V/V) acetonitrile and 30 mmol/LSDS, under 25 kV applied voltage and at 25 degrees C. The linear range, the limit of detection and the repeatability were investigated under the optimized conditions. The RSDs (n = 6) of the migration time and peak area for each nucleotide are less than 1% and 3%, respectively.