Drift tube; Multiplexing; Oligonucleotides; Mass Spectrometry; Ion Mobility Spectrometry; Collision cross sections; Diastereomers
Abstract :
[en] Hydrophilic interaction liquid chromatography (HILIC) coupled to drift tube ion mobility spectrometry (DTIMS) was used to separate diastereomers of five-unit oligonucleotides containing 0, 1, 2 or 3 phosphorothioate (PS) linkages. Multiplexed DTIMS (where ions are pulsed into the drift tube according to a pre-encoded sequence) and post-acquisition processing using an innovative demultiplexing tool were investigated. The electric field inside the drift tube was optimized to achieve the highest resolving power. The entrance voltage providing the best two-peak resolution was -1000V with 3-bit multiplexing. Under optimized conditions, the eight diastereomers of an oligonucleotide with three PS linkages (5'-TC∗G∗T∗G-3') could be separated unambiguously. Indeed, those diastereomers differed in their collision cross section (CCS) values. The minimal CCS values difference between two adjacent diastereomers was 0.9% with maximal RSD on CCS values of 0.3%. The use of multiplexed ion mobility and the novel high-resolution demultiplexing tool represents a real breakthrough for resolution enhancement of diastereomers in linear DTIMS.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Demelenne, Alice ; Université de Liège - ULiège > Université de Liège - ULiège
Nys, Gwenael ; Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liege, Quartier Hôpital, Avenue Hippocrate 15, 4000, Liege, Belgium
Nix, Cindy ; Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Fjeldsted, John C; Agilent Technologies, Santa Clara, CA, 95051, United States
Crommen, Jacques ; Université de Liège - ULiège > Département de pharmacie
Fillet, Marianne ; Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Separation of phosphorothioated oligonucleotide diastereomers using multiplexed drift tube ion mobility mass spectrometry.
Alice Demelenne: experiments, redaction. Gwenael Nys: support for the experiments relative to IMS and redaction reviewing. Cindy Nix: support for the experiments relative to IMS. John C. Fjeldsted: support for the experiments relative to IMS. Jacques Crommen: redaction reviewing. Marianne Fillet: study design, financial support, Supervision, redaction reviewing.
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