Computer-assisted retention prediction system for oligonucleotides in gradient anion-exchange chromatography

A technique is presented for the retention prediction of single-stranded homo-oligonucleotides under gradient elution conditions in anion-exchange chromatography. The prediction system is based on the theory proposed by Jandera and Churacek. In the present system, the theory was modified by the extrapolation method, utilizing the linear relationship between the log of the capacity factor and the number of nucleotides. The modified theory allowed the prediction of retention times for high-molecular-weight oligonucleotides, which could not be calculated from the original theory. Oligoadenylate was used as a standard oligonucleotide to demonstrate the accuracy of the prediction system. By use of this system, the retention times of oligoadenylates up to 25-35 bases in chain length were predicted within 8% errors under both binary-linear and binary-convex gradient shapes.