使用可变数据独立获取用于基于毛细管电泳的非目标代谢学.
Saki Kiuchi1, Yasuhiro Otoguro2, Tomoaki Nitta2
1Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Journal of the American Society for Mass Spectrometry
|August 13, 2024
概括
使用毛细电泳-并联质谱 (CE-MS/MS) 的可变数据独立获取 (vDIA) 通过提高光谱质量和注释率来改善代谢分析. 这个优化的平台准确地识别复杂的生物样本中的代谢物,包括新型代谢物.
科学领域:
- 分析化学 分析化学
- 代谢学 代谢学 代谢学
- 生物化学 生物化学
背景情况:
- 毛细管电泳与双重质谱学 (CE-MS/MS) 结合,为代谢分析提供了高峰容量和灵敏度.
- 在MS/MS中数据独立获取 (DIA) 在分析化学和信息学方面存在限制,以实现最佳的代谢分析.
- 现有的DIA方法缺乏标准化程序,阻碍了全面的代谢分析.
研究的目的:
- 在CE-MS/MS中评估和比较全离子碎片 (AIF) 和可变DIA (vDIA) 技术的质谱质量.
- 开发和验证一个优化的信息学框架,以使用CE-MS/MS进行增强的代谢分析.
- 使用开发的平台,研究脂聚糖 (LPS) 诱导的巨细胞中的代谢变化.
主要方法:
- 评估了前体离子 (60-800 Da) 隔离的 AIF 和 vDIA 技术,并评估了光谱质量和注释率.
- 实施了使用内部标准进行精确峰值对齐的线性迁移时间 (MT) 校正方法.
- 使用MS-DIAL,MS-FINDER和分子频谱网络进行数据分析,峰值注释和结构阐明.
主要成果:
- 与MS-DIAL算法相结合,vDIA与AIF相比显示出更高的光谱匹配得分和注释率.
- MT校正方法实现了不到0.1分钟的MT漂移,相当于逆相液态染色学.
- 一个大规模的MT预测 (469,870种化合物) 实现了<1.5分钟的平方根平均精度;在LPS诱导的巨细胞中表征了170种代谢物,包括新发现的甘氨胺核酸.
结论:
- 优化的CE-MS/MS与VDIA和先进的计算工具提供了一个强大的平台,用于高通量代谢分析.
- 这种方法提高了代谢物识别的准确性,并使以前没有注释的或新型化合物的特征成为可能.
- 该研究强调了CE-MS/MS-DIA和计算质谱学的潜力,以推动代谢学研究.
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