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建立O标记的伊诺酸用于定量毛细管电泳-质谱:碎片化途径和与C标记的类似物进行比较

Guizhen Liu^1,2, Tobias Dürr-Mayer^1,2, Mengsi Lu¹

¹Institute of Organic Chemistry, University of Freiburg, Albertstrasse 21, 79104 Freiburg, Germany.

Analytical chemistry

|November 5, 2025

概括

标有Oxygen-18的内醇酸盐 (InsPs) 和热酸盐 (PP-InsPs) 为毛细电泳质谱法 (CE-MS) 量化提供了碳-13标准的经济有效替代品. 这种方法为复杂的细胞提取物提供了准确和可重复的结果.

科学领域:

分析化学分析化学
生物化学生物化学
分子生物学分子生物学

背景情况:

毛细电泳质谱 (CE-MS) 对于量化内醇酸盐 (InsPs) 和内醇热酸盐 (PP-InsPs) 是至关重要的.
新型InsP和PP-InsP异构体的发现需要新的重同位素标记的内部标准来准确分析复杂的生物样本.
目前的方法通常依赖于13C标记的标准,这些标准可能是昂贵和复杂的合成.

研究的目的:

评估18O标记的InsPs和PP-InsPs作为13C标记的CE-MS量化内部标准的负担得起和可访问的替代品.
为了研究MS2碎片化路径和18O标记化合物的同位素再分配.
通过使用18O标记的标准,确定最佳的碎片化过渡以进行可靠的定量分析.

主要方法:

合成各种18O标记的InsPs和PP-InsPs,其18O原子的数量和位置不同.
系统地研究MS2碎片化路径,以了解18O再分配.
应用CE-MS用于用18O标记标准对细胞提取物进行定量分析,并与基于13C的方法进行比较.

主要成果:

为18O标记的InsPs和PP-InsPs提出了两个主要的分离途径,阐明了18O再分配.
鉴定出HPO3的损失是一个合适的MS2过渡,它最大限度地减少同位素再分配,提供可重现的定量比率.
在分析来自Saccharomyces cerevisiae,HCT116细胞和Arabidopsis thaliana的提取物时,获得了与基于13C的方法可比的准确量化和精度.

结论:

18O标记的InsP和PP-InsP是CE-MS量化13C标记标准的可行,经济高效的替代品.
了解碎片化途径和选择适当的产物离子 (例如,HPO3的损失) 是可靠量化的关键.
这种方法使复杂细胞提取物的精确和准确分析成为可能,促进对生物系统中InsP和PP-InsP作用的研究.

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