Development of a pseudotargeted metabolomics approach for relative quantification of free fatty acid double-bond isomers in beagle plasma

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2025-08-21 DOI:10.1016/j.jchromb.2025.124765

Wen Xiao , Haihui Ren , Yueyi Zhang , Jiameng Han , Qiaorong Liu , Zunjian Zhang , Yuan Tian

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Abstract

Free fatty acids (FFAs) play a key role in living organisms and participate in metabolic processes, mainly as a source of energy for cells. The CC position isomer of FFAs is strongly associated with many diseases, but quantification of its double bond position isomer remains a challenge in lipid analysis. In this study, based on the requirements of the pseudotargeted metabolomics, an analytical method based on a two-step derivatization LC-MS/MS method and a multiple reaction monitoring (MRM) mode was established and fully validated in terms of linearity, precision, and stability. The results demonstrated that the method relatively quantified 30 FFAs in plasma samples. These included 8 saturated fatty acids (SFAs), 12 monounsaturated fatty acids (MUFAs), and 10 polyunsaturated fatty acids (PUFAs). The developed method was applied to analyze the increase or down-regulation of the relative content of FFAs before and after the administration of the drug in beagles which may help to explain the therapeutic effect of sacubitril valsartan. The establishment of this method allows the study of FFAs in beagles plasma to be no longer limited to analysis at the subclass level but can be focused at the isomer level.

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小猎犬血浆中游离脂肪酸双键异构体相对定量的假靶向代谢组学方法的发展

游离脂肪酸（FFAs）在生物体中发挥关键作用，参与代谢过程，主要是作为细胞的能量来源。FFAs的CC位置异构体与许多疾病密切相关，但其双键位置异构体的定量在脂质分析中仍然是一个挑战。本研究根据伪靶向代谢组学的要求，建立了一种基于两步衍生化LC-MS/MS法和多反应监测（MRM）模式的分析方法，并对其线性度、精密度和稳定性进行了充分验证。结果表明，该方法可相对定量血浆样品中的30种游离脂肪酸。其中包括8种饱和脂肪酸（sfa）， 12种单不饱和脂肪酸（mufa）和10种多不饱和脂肪酸（pufa）。应用所建立的方法分析给药前后小猎犬体内游离脂肪酸相对含量的升高或下调，有助于解释苏比利缬沙坦的治疗效果。该方法的建立使得对比格犬血浆中游离脂肪酸的研究不再局限于亚类水平的分析，而可以集中在同分异构体水平。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.