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首页> 外文期刊>RSC Advances >A rapid and sensitive profiling of free fatty acids using liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) after chemical derivatization
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A rapid and sensitive profiling of free fatty acids using liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) after chemical derivatization

机译：使用液相色谱电离电离串联质谱（LC / ESI-MS / MS）在化学衍生化之后的快速和敏感的游离脂肪酸分析

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摘要

Free fatty acids (FFAs) have diverse roles in cellular energy and signaling and they are critical molecules in various biological states. Due to the poor ionization efficiency of FFAs under electrospray ionization mass spectrometry (ESI-MS) conditions, it is a challenging aspect to construct a robust platform for profiling of various FFAs in biological samples using liquid chromatography ESI-MS. In the present study, we applied trimethylsilyldiazomethane (TMSD) derivatization to improve ionization efficiencies in the profiling of FFAs. Multiple reaction monitoring (MRM) was used for the selective quantification of methylated FFAs. The optimal TMSD methylation was validated for a reliable FFA profiling. Furthermore, the high-throughput analysis of FFAs was successfully performed in short analysis and derivatization times. To verify the utility and effectiveness of the developed method, we compared both methylation and nonmethylation (intact FFA) data in the profiling of FFAs in mice liver and plasma. It is noteworthy that the methylation derivatization provided better results in FFA profiling. Further, we performed statistical data analysis where HBV and mock mice tissues were discriminated when the methylated FFAs data were used. In the lipidomics field, the present method can also be applied for the profiling of FFAs in biological samples for biomarker discovery. The present validated LC/ESI-MS/MS assay method may also be used for FFA profiling modeling studies in other biomedical samples.
机译：游离脂肪酸（FFA）具有多种细胞能量和信号传导的作用，并且它们是各种生物态的关键分子。由于电喷雾电离质谱法（ESI-MS）条件下FFA的电离效率差，它是一种具有挑战性的方面，用于构建使用液相色谱ESI-MS在生物样品中的各种FFA的勘探平台的鲁棒平台。在本研究中，我们应用三甲基甲硅烷二甲基甲烷（TMSD）衍生化，以改善FFA的分析中的电离效率。多重反应监测（MRM）用于选择性定量甲基化FFA。验证了最佳TMSD甲基化以获得可靠的FFA分析。此外，在短期分析和衍生时间内成功进行了FFA的高通量分析。为了验证开发方法的实用性和有效性，我们将甲基化和非甲基化（完整的FFA）数据与小鼠肝脏和血浆中FFA的分析中的差异化。值得注意的是，甲基化衍生化在FFA分析中提供了更好的结果。此外，我们进行了统计数据分析，其中当使用甲基化的FFA数据时，区分HBV和模拟小鼠组织。在脂质体域，本方法也可以应用于生物标志物发现的生物样品中FFA的分析。本验证的LC / ESI-MS / MS测定方法也可用于其他生物医学样品中的FFA分析建模研究。

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