Abstract
There has been a significant increase in the use of hydrophilic interaction liquid chromatography (HILIC) to separate oligonucleotides. This rise in the use of HILIC has correlated to the increasing success of oligonucleotides as therapeutic treatments and reagents in biomedical research. As more scientists need to routinely analyze oligonucleotides in addition to small molecules, peptides, and proteins using the same analytical instruments, it becomes difficult to use traditional types of analyses such as ion-pair reversed-phase chromatography. This increased use has led to new approaches that have improved the utility of HILIC to the point where it has become a legitimate alternative approach to ion-pair reversed-phase chromatography. This review highlights recent advances in HILIC separations of oligonucleotides with a focus on the underlying mechanisms of action. While HILIC has made significant gains in performance, there still remain challenges, which if properly addressed will continue to propel this approach forward.
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