Analytical Sciences

Ion-pair Reversed-phase Liquid Chromatographic Separation of Oligonucleotides

Takumi TAKANO,* Chiaki AOYAMA,* Yoshiro TERASAKI,* Kenichi SUZUKI,* Aki ANDO,* Yanting SONG,** and Makoto TSUNODA***

*GL Sciences Inc., 6-22-1 Nishi Shinjuku, Shinjuku, Tokyo 163-1130, Japan
**School of Pharmaceutical Sciences, Hainan University, Haikou 570228, China
***Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033, Japan

Therapeutic oligonucleotides have recently been approved in the United States, the EU, and Japan. Hence, the analysis of oligonucleotides is an important topic in drug development. Liquid chromatographic techniques are commonly used for purity verification and the determination of oligonucleotides. In ion-pair reversed-phase separation, several parameters, such as the pore size of the stationary phase, mobile phase additives, and column temperature, were investigated using three types of oligonucleotides (18, 19, and 20 mer). All of the investigated parameters could influence the separation, and they are expected to be useful for optimizing oligonucleotide separation.

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