This study evaluated four bridged‐ethylene hybrid (BEH) columns containing C₁₈ (130 Å), peptide C₁₈ (300 Å), phenyl, or a mixed‐mode charged surface hybrid (CSH C₁₈) using a wide range of antisense oligonucleotide therapeutics. The BEH C₁₈, 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 C₁₈, 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 C₁₈ column provided the best peak shape. The CSH C₁₈ 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.

中文翻译：

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评估烷基胺和固定相以改善寡核苷酸的LC-MS

这项研究评估了四个桥联乙烯杂化（BEH）色谱柱，其中包含C ₁₈（130Å），肽C ₁₈（300Å），苯基或使用多种反义寡核苷酸的混合模式带电表面杂化（CSH C ₁₈）疗法。BEH C ₁₈，肽和苯基色谱柱均能够在使用烷基胺和1,1,1,3,3,3，-六氟异丙醇（HFIP）的多个离子对系统中显着保留寡核苷酸样品。寡核苷酸的保留根据烷基胺的选择而变化，保留的顺序为二甲基环己胺>二异丙基乙胺>三乙胺。这些色谱柱对几种紧密洗脱的杂质的选择性相似。虽然总体上C_在图18中，发现肽和苯基柱均能够分析寡核苷酸治疗剂，发现苯基柱具有最大的保留性，而C ₁₈柱则提供了最佳的峰形。尽管流动相在色谱柱的pH稳定范围内，但发现CSH C ₁₈色谱柱会被烷基胺-HFIP流动相降解。