使用重原子同位素标记为l-和d-Marfey的试剂的立体选择性氨基组学.
Nitish R Mishra1, William G Gutheil1
1Division of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Missouri─Kansas City, Kansas City, Missouri 64108, United States.
Journal of the American Society for Mass Spectrometry
|April 29, 2024
概括
一种新方法使用重标记的马菲试剂进行精确的生物胺和氨基酸的立体异构体分析. 这种技术通过改进性化合物的识别和量化来增强"氨基组学"研究.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 代谢学 代谢学 代谢学
背景情况:
- 生物氨基和氨基酸对于许多生化过程至关重要.
- 对"氨基组学"来说,分析性氨基和氨基酸的立体异构体至关重要,但由于性分离方法的局限性而具有挑战性.
- 之前的工作确立了马菲的试剂,通过LC-MS/MS通过氨基和氨基酸立体异构体进行溶解.
研究的目的:
- 开发一种重原子同位素标记的马菲试剂方法,用于对胺和氨基酸的立体选择检测和定量.
- 通过实现精确的立体异构体识别来增强"氨基组学"的分析能力.
主要方法:
- 重型 (13C2) l-Marfey's (Hl-Mar) 和重型 (2H3) d-Marfey's (Hd-Mar) 试剂的合成. 这两种试剂的组合过程中,重型 (13C2) l-Marfey's (Hl-Mar) 和重型 (2H3) d-Marfey's (Hd-Mar) 试剂的合成过程中,重型 (13C2) l-Marfey's (Hl-Mar) 和重型 (2H3) d-Marfey's (Hd-Mar) 试剂的合成过程中使用的试剂.
- 使用轻型和重型马菲试剂来衍生氨基混合物.
- 通过液体染色学四极点飞行时间高分辨率质谱法 (LC-QToF-HRMS) 进行分析.
- 对质量差异的比较分析 (轻与重的马菲 adducts) 和保留时间来识别立体同位素.
主要成果:
- 成功合成了重标记的马菲试剂 (Hl-Mar和Hd-Mar).
- 根据质量差异,证明了识别单元,二元和三元马菲 adducts的能力.
- 通过l-和d-Mar衍生物之间的保留时间差异来准确识别立体同位素.
- 该方法的应用,以识别甲素耐药黄金葡萄球菌 (MRSA) 提取物中的奇拉和阿奇拉成分.
结论:
- 重原子同位素标记的马菲试剂方法为氨基和氨基酸立体异构体分析提供了高的分析选择性和可重复性.
- 这种方法通过使精确的立体异构体量化,显著提高了"氨基组学"研究的能力.
- 开发的技术是强大的,适用于复杂的生物样本.
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