在没有离子配对试剂的情况下进行寡核酸分析的基静止相的系统评估
Athanasios Tsalmpouris1, Eric Largy2, Davy Guillarme1
1School of Pharmaceutical Sciences, University of Geneva, CMU - Rue Michel Servet 1 1211, Geneva, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, CMU - Rue Michel Servet 1 1211, Geneva, Switzerland.
Journal of chromatography. A
|October 2, 2025
概括
研究人员探索了基于的静止相,作为对离子对逆相液态色谱 (IP-RPLC) 的替代方法,用于分析寡核酸 (ON). 双和五烯 (PFP) 柱显示出有前途,提供无离子配对试剂的MS兼容分离.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 药品分析 药品分析
背景情况:
- 寡核酸 (ONs) 是重要的治疗药物,但由于电荷,修饰和杂质,它们的分析具有挑战性.
- 目前的黄金标准,离子对逆相液态色谱 (IP-RPLC),需要不良的离子对应试剂.
- 开发无离子对的方法对于强大的ON表征至关重要.
研究的目的:
- 评估基于的静止相作为IP-RPLC用于寡核酸 (ON) 分离的替代品.
- 研究控制ON与基相相互作用的保留机制.
- 证明基于的RPLC用于分析治疗性ONs的适用性.
主要方法:
- 系统选九个基改性静止相 (基-基,二基,二基,PFP,五基基).
- 优化色谱条件 (移动阶段,温度,梯度,流速).
- 使用量子力学建模和循环二元论来阐明保留机制的补充研究.
主要成果:
- 双和五烯 (PFP) 柱显示出优越的峰值容量和对ONs的选择性.
- 使用50mM氨酸 (pH 8),甲醇,50°C和20分钟梯度实现了最佳分离.
- 保留主要是由π-π堆叠驱动的,受ON二次结构和甲醇度的影响.
- 基于的RPLC表现出与IP-RPLC可比的峰值容量,并且与MS兼容.
结论:
- 基于的静态相,特别是双和PFP,为寡核酸分析提供了可行的,无离子对的替代方案.
- 这些方法是强大的,MS兼容,并补充传统的IP-RPLC.
- 这种方法扩大了用于表征治疗性寡核酸的分析能力.
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