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Evaluation of alkylamines and stationary phases to improve LC–MS of oligonucleotides
Author(s) -
Bartlett Michael G.,
Omuro Shogo
Publication year - 2021
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.5045
Subject(s) - chemistry , oligonucleotide , triethylamine , chromatography , selectivity , peptide , phase (matter) , combinatorial chemistry , impurity , organic chemistry , catalysis , dna , biochemistry
Abstract This study evaluated four bridged‐ethylene hybrid (BEH) columns containing C 18 (130 Å), peptide C 18 (300 Å), phenyl, or a mixed‐mode charged surface hybrid (CSH C 18 ) using a wide range of antisense oligonucleotide therapeutics. The BEH C 18 , peptide, and phenyl columns were all capable of providing significant retention of oligonucleotide samples across multiple ion‐pairing systems using alkylamines and 1,1,1,3,3,3,‐hexafluoroisopropanol (HFIP). The retention of the oligonucleotides varied depending on the choice of alkylamine, with the order of retention being dimethylcyclohexylamine > diisopropylethylamine > triethylamine. The selectivity of these columns for several closely eluting impurities was similar. Although overall the C 18 , peptide, and phenyl columns were all found to be capable of analyzing oligonucleotide therapeutics, the phenyl column was found to be the most retentive and the C 18 column provided the best peak shape. The CSH C 18 column was found to be degraded by the alkylamine‐HFIP mobile phase despite the mobile phase being within the pH stability range of the column.