Separation/analysis method for mixture of oligonucleotides
The present invention relates to a method of separating and analyzing a mixture of oligonucleotides, including performing liquid chromatography using a column packed with a packing material obtained by fixing a diol to a surface of each of porous particles formed of a crosslinked organic polymer. According to this method, the oligonucleotides can be separated and analyzed with higher sensitivity compared to cases where columns having silica gel as a base material are used. In addition, the column can be washed with an alkaline solution.
1. A method of separating and analyzing a mixture of oligonucleotides, including performing liquid chromatography using a column packed with a packing material obtained by fixing a diol to a surface of each of porous particles consisting of a crosslinked organic polymer selected from the group consisting of a (meth)acrylic resin and a crosslinked polyvinyl alcohol,
wherein a structure of the diol includes a structure represented by the following formula (I):
R—O—CH 2 —CH(OH)—CH 2 (OH) (I)
where R represents a partial structure of each of the porous particles, and
wherein R consists of a crosslinked organic polymer selected from the group consisting of a (meth)acrylic resin and a crosslinked polyvinyl alcohol.
2. The method according to claim 1 , wherein the crosslinked organic polymer is crosslinked polyvinyl alcohol.
3. The method according to claim 1 , wherein a mobile phase of the liquid chromatography is a mixed liquid of an aqueous solution of a volatile salt and a water-soluble organic solvent.
4. The method according to claim 1 , further including detecting an eluted component separated through the column with an ultraviolet/visible detector and/or a mass spectrometer.
5. The method according claim 1 , wherein the liquid chromatography is performed by gradient elution of a mobile phase.
6. The method according claim 1 , further including, before or after the performing liquid chromatography, a step of washing the column by passing an alkaline solution having a pH of from 10.0 to 13.0 through the column.
7. The method according to claim 1 , wherein the oligonucleotides are synthetic oligonucleotides.
8. The method according to claim 6 , wherein the crosslinked organic polymer is crosslinked polyvinyl alcohol.