开发多重切割心脏的二维液态色谱,在两个维度中进行离子配对逆相分离,用于分析治疗性寡核酸中的杂质
Dwight Stoll1, Maria Sylvester1, Daniel Meston1
1Department of Chemistry, Gustavus Adolphus College, Saint Peter, MN 56082, USA.
Journal of chromatography. A
|December 16, 2023
概括
二维液态色谱 (2D-LC) 为分析复杂的寡核酸杂质提供了强大的解决方案. 本研究提出了一种代方法开发方法,用于在2D-LC.中快速,高选择性梯度生成.
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:
- 由于其复杂性,寡核酸存在重大生物分析挑战.
- 一维 (1D) 液态染色学 (LC) 方法不足以解决寡核酸样本中的所有杂质.
- 二维液态色谱 (2D-LC) 为复杂的混合物提供了增强的分离能力.
研究的目的:
- 开发一种简单的2D-LC方法,用于快速生成高选择性梯度.
- 应用一个代的保留建模方法,以优化寡核酸杂质分析.
- 为了证明在制药性寡核酸样本中阐明关键杂质对.
主要方法:
- 离子配对逆相 (IPRP-IPRP) 2D-LC方法的开发.
- 使用两次训练运行进行代梯度生成,以预测在较浅的梯度中的保留.
- 在第二维中应用量身定制的高选择性梯度,用于关键对的分离.
主要成果:
- 从训练运行中成功生成12%梯度的代生成,预测在2%梯度中保持低误差 (3%和11%).
- 对于全长产品和酸杂质,实现了低至1:1000的杂质概况.
- 在复杂的原始制药性寡核酸样本中证明有效地阐明关键杂质对.
结论:
- 代保留建模方法使复杂的寡核酸分离能够快速和轻松地开发2D-LC方法.
- 2D-LC是解决寡核酸杂质带来的生物分析挑战的强大工具.
- 这种方法有助于改善药品寡核酸产品的质量控制和表征.
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