Journal
ANALYTICAL BIOCHEMISTRY
Volume 625, Issue -, Pages -Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2021.114194
Keywords
Oligonucleotide; Ion-pairing reagent; Core-shell; Liquid chromatography mass spectrometry; Fully porous
Funding
- Natural Sciences Foundation of Guangdong Province [2019A1515012078]
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In this study, the correlation among stationary phases, ion-pairing reagents, and sequences for the analysis of oligonucleotide (ODN) using ion-pair reversed-phase liquid chromatography mass spectrometry was evaluated with C18 columns. It was found that silica-based porous particles retained ODN the strongest, while hexylamine produced the longest retention for all ODNs among the six ion-pairing reagents. Additionally, the separation ability of C18 columns depends on the type of ion-pairing reagent and ODN sequences, with little impact of particle type on signals of ODNs for most synthetic sequences.
The correlation among stationary phases, ion-pairing reagents (IPR) and sequences for ion-pair reversed-phase liquid chromatography mass spectrometry (IP-RP LC-MS) analysis of oligonucleotide (ODN) remains unclear. The present study aimed to evaluate such correlation using particle-packed C18 columns in order to search for the optimal combination among them. Five C18 columns packed with core-shell silica, polymer, porous silica and hybrid particles, respectively, were evaluated for the analysis of synthetic and chemically modified ODNs with six different IPRs. Our results showed that silica-based porous particles, compared to other particles, retained ODN the strongest no matter which IPR was used. Meanwhile, among the six IPRs hexylamine (HA) produced the longest retention for all ODNs, regardless of the types of C18 particles. For the separation of ODNs, C18 columns performed similarly under identical LC conditions. However, the separation ability of C18 columns is highly dependent on the type of IPR and ODN sequences. Moreover, the type of particles has little impact on the signals of ODNs for the majority of synthetic sequences, but such impact could be dramatic for chemically modified sequences. On the other hand, both the type of IPR and ODN sequence have a significant effect on MS signals for synthetic and chemically modified ODNs.
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