Study on Retention Behaviour of Homo-Oligonucleotides in IP-RPLC Using Dual-Point Retention Time Correction on 'Standard Column' with Internal Standards

摘要

The retention behavior of homo-oligonucleotides was investigated in ion-pair reverse-phase high-performance liquid chromatography (IP-RPLC). Rectification of retention time was performed by introducing the concept of "standard column" with double-point retention time correction (DP-RTC) using two anchor oligonucleotides as internal standards to ensure the accuracy of the retention measurement. Through polynomial fitting, a quadric relationship was adopted between normalized retention time and chain length of (dA)_(10-21) and (dT)_(10-21). The retention times of homo-oligonucleotides depended on the number of bases and increased with increasing hydrophobicity, namely, A