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通过方便和定量甲基化策略,扩大多段注射-非水性毛细管电泳-质谱的脂质覆盖范围

Ritchie Ly¹, Lucas Christian Torres¹, Nicholas Ly¹

¹Department of Chemistry and Chemical Biology, McMaster University, 1280 Main Street West, Hamilton, Ontario, Canada L8S 4M1.

Analytical chemistry

|November 22, 2023

概括

一种新方法使用一种更安全的衍生协议,通过毛细管电泳质谱分析进行脂分析. 这种技术改善了脂层层覆盖范围,为复杂的生物样本提供了更高的分辨率,灵敏度和吞吐量.

科学领域:

分析化学分析化学
生物化学生物化学
质谱测量质量谱测量
利皮多米克 (Lipidomics) 是一种消化剂.

背景情况:

描述复杂的人类脂质组需要先进的分离技术与高分辨率质谱相结合.
与染色体和离子流动性方法相比,电泳分离在脂管学中未得到充分利用.
现有的方法面临着异干扰和离子抑制的挑战,限制了全面的脂管组分析.

研究的目的:

为量化脂分析引入一种新的,更安全的衍生协议.
为了使脂的快速分析使用多段注射-非水性毛细血管电泳-质谱 (MSI-NACE-MS).
扩大脂质组覆盖范围,提高复杂生物样本的分析性能.

主要方法:

开发了一种使用3 - 甲基-1-p - 托利特里亚 (MTT) 的衍生协议,作为脂甲基化中对二甲的更安全替代品.
实施了充电开关衍生策略,涉及9 - 烯甲基氧碳化物和逆提取,以最大限度地减少干扰.
使用MSI-NACE-MS分析人体血提取物,对已建立的脂管学数据和参考材料进行方法验证.

主要成果:

实现了高效的定量脂甲基化 (∼90%) 和快速样本分析 (∼3.5分钟/样本).
证明了更好的分辨率,灵敏度和吞吐量,特别是对于分析为酸甲基的酸.
在人体血中成功比较了甲基和甲基物种,与实验室间研究结果保持一致,并使用NIST SRM-1950显示了量化潜力.

结论:

基于MTT的衍生协议通过MSI-NACE-MS为脂分析提供了一种实用且更安全的方法.
这种充电切换策略显著扩大了脂质组覆盖范围,增强了脂质组学中的毛细血管电泳的功能.
该方法为定量脂管学提供了一个强大的平台,通过电泳性移动性建模和MS/MS支持脂质识别.

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