Twin Derivatization Strategy for High-Coverage Quantification of Free Fatty Acids by Liquid Chromatography-Tandem Mass Spectrometry
Twin Derivatization Strategy for High-Coverage Quantification of Free Fatty Acids by Liquid Chromatography-Tandem Mass Spectrometry
复制标题
通过液相色谱-串联质谱法高覆盖度定量游离脂肪酸的双衍生化策略
DOI：
10.1021/acs.analchem.7b03020
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发表时间：
2017-11-21
影响因子：
7.4
通讯作者：
Xu, Fengguo
中科院分区：
文献类型：
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作者：
Jiang, Ruiqi;Jiao, Yu;Xu, Fengguo
游离脂肪酸（Free Fatty Acids，FFA）是脂质的重要组成部分，以多种方式调节生物代谢。尽管游离脂肪酸的分子结构简单，但由于其独特的性质和广泛的浓度范围，其分析仍然具有挑战性。建立了高覆盖度液相色谱-串联质谱（LC-MS/MS）法测定血清样品中游离脂肪酸的方法，采用两种结构类似物5-二甲氨基萘磺酰哌嗪（Dns-PP）和二乙氨基萘磺酰哌嗪（Dens-PP）作为双衍生试剂。Dns标记的游离脂肪酸可以显着提高其MS响应，通过引入容易电离的部分，含叔胺的部分，并帮助在多反应监测（MRM）模式的碎片。结果表明，与非衍生化方法相比，FFAs的检测灵敏度提高了50-1500倍。同时，使用Dens标记的标准品作为一对一的内标，以确保准确定量。涵盖短链、中链和长链的38种游离脂肪酸可在宽动态范围内定量，定量下限（LLOQ）为2至20 nM。使用这种方法，我们分别分析了顺铂诱导的肾毒性和伊立替康诱导的胃肠道毒性大鼠模型中的血清FFAs。这些发现与先前非靶向代谢组学所揭示的结果进行了进一步比较。结果表明，基于孪生衍生的LC-MS提供了更准确的全局FFA变化的视图，并在靶向代谢组学领域具有巨大的应用潜力。
Free fatty acids (FFAs) are vitally important components of lipids that modulate biological metabolism in various ways. Although the molecular structures are simple, the analysis of FFAs is still challenging due to their unique properties and wide concentration range. In the present study, a high-coverage liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the quantification of FFAs in serum samples using two structural analogues 5-(dimethylamino)naphthalene-1-sulfonyl piperazine (Dns-PP) and (diethylamino)naphthalene-1-sulfonyl piperazine (Dens-PP) as twin derivatization reagents. The Dns labeling of FFAs could significantly enhance their MS response via the introduction of the easily ionizable moiety of a tertiary amine-containing part and aid fragmentation in the multiple reaction monitoring (MRM) mode. Our results demonstrated that the detection sensitivities of FFAs were increased by 50-1500 fold compared with the nonderivatization method. At the same time, Dens-labeled standards were used as one-to-one internal standards to ensure accurate quantifications. Thirty-eight FFAs, covering short-, medium-, and long-chain, could be quantified in wide dynamic range with the lower limit of quantification (LLOQ) varied from 2 to 20 nM. Using this method, we analyzed serum FFAs in rat models of cisplatin-induced nephrotoxicity and irinotecan-induced gastrointestinal toxicity, respectively. The findings were further compared with those revealed by previous untargeted metabolomics. The results indicate that twin derivatization-based LC-MS provides a more accurate view of global FFA alternation and has great application potential in the fields of targeted metabolomics.