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开发一种基于水友互动的方法,用于提取,分离和确定治疗性寡核酸

Zuzana Vosáhlová¹, Szymon Bocian¹, Sylwia Studzińska¹

¹Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin Str., PL-87-100, Toruń, Poland.

Analytica chimica acta

|November 24, 2025

概括

这项研究开发了一种简单的,可重复的方法来提取和分析血清中的改性寡核酸,使用水友相互作用液态色谱. 新方法实现了高回收率,这对于精确的质谱检测寡核酸杂质和代谢物至关重要.

科学领域:

分析化学分析化学
生物化学生物化学

背景情况:

精确分析血清中的寡核酸杂质和代谢物,需要可靠的提取和质谱检测.
具有质谱学兼容的移动相的水友交互液态色谱 (HILIC) 是这种分析的理想选择.
现有的提取方法可能缺乏复杂生物样本所需的重复性和回收性.

研究的目的:

开发一种简单且可重复的方法来提取和分析不同长度 (4mer-14mer) 的改性寡核酸在血清中.
为提取和染色学分析优化水友相互作用条件.
为了实现高回收率,并确保与质谱检测的兼容性.

主要方法:

在不同的移动相组合和温度下,在zwiwterionic和amide柱上进行了一项全面的保留研究.
质谱参数被优化为最大的灵敏度,解决 adduct 形成的挑战.
一种新合成的胺基吸附剂被用于从丰富的血清提取物中分散固相提取 (dSPE).
基于pH驱动的弱离子交换机制,优化了dSPE的吸附和脱离条件.

主要成果:

胺基列被选择而不是基列,因为它的稳定性和能够化较长的寡核化物 (>10mer).
优化的质谱参数提高了灵敏度.
使用胺基吸附剂开发的dSPE方法实现了高回收率 (>75%) 和优异的重复性 (RSD <5%) 对于血清提取物中修饰的寡核酸.

结论:

基于完全水性相互作用的程序首次成功地用于提取和分析血清中修饰的寡核酸混合物.
该方法表现出简单性,可重复性和高回收率,使其适合分析寡核酸杂质和代谢物.
这一进步有助于使用质谱仪在复杂的生物基质中更可靠,更灵敏地检测寡核化物.

关键词:

氨基酸吸附剂是一种氨基酸吸附剂.分散固体相提取方法分散固体相提取方法水友互动液体色谱学液体色谱学质谱测量质量谱测量修改后的反意义寡核酸.

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