RAPID ANALYSIS OF RIBONUCLEOSIDES AND BASES AT PICOMOLE LEVEL USING PELLICULAR CATION EXCHANGE RESIN IN NARROW BORE COLUMNS

The separation of the four major bases and nucleosides of RNA has been investigated under conditions utilizing wide ranges of pH, temperature, and flow rates. Smallbore columns packed with pellicular cation-exchange resin were used in a high-pressure liquid chromatograph equipped with a micro UV detector. Dilute acidic potassium or ammonium phosphate solutions served as eluents. The four bases or nucleosides were analyzed at the subnanomole level in less than 6 minutes using 3-meter colunms and high-flow velocities. With a shorter column operated at relatively low flow velocities, a few picomoles of ribonucleosides can be separated and determined without resorting to radioactive techniques. © 1969, American Chemical Society. All rights reserved.