Reversed-phase high-performance liquid chromatography of protamines

The protamines from the gonads of the sturgeonAcipenser stellatus have been separated by high-performance liquid chromatography. The proteins were eluted with mixtures of water and ethanol having a gradient of ethanol concentrations in the presence of trifluoroacetic acid (TFA). The influence of the concentration of TFA and the temperature of the column on separation was studied. The quantitative (95–98%) isolation of the protamines from the column was achieved at a temperature of 30°C and a 0.15% concentration of TFA.