The optimization and validation of a gas chromatography-mass spectrometry method to analyze the concentration of acetate, propionate and butyrate in human plasma or serum

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2024-09-07 DOI:10.1016/j.jchromb.2024.124299

Greet Vandermeulen, Riet Rosseel , Georgia Chatonidi, Lise Deroover , Eef Boets , Kristin Verbeke

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引用次数: 0

Abstract

Fermentation-derived short-chain fatty acids (SCFA)⁴ are potential mediators of the health benefits associated with dietary fiber intake. SCFA affect physiological processes locally in the gut and on distant organs via the systemic circulation. Since SCFA are used as energy source for colonocytes and substrate for the liver metabolism, their concentrations in the systemic circulation are low. Therefore, quantification of systemic SCFA requires sensitive analytical techniques. This article covers the optimization and validation of a gas chromatography-mass spectrometry method to measure systemic SCFA concentrations following derivatization with 2,4-difluoroaniline (DFA)⁵ and extraction in ethyl acetate. Sample preparation was optimized by varying the amount of DFA, coupling agent 1,3-dicyclohexylcarbodiimide, ethyl acetate and sodium bicarbonate, which is used to quench derivatization. In addition, evaporation of the samples using a vacuum concentrator resulted in less contamination, notably of acetate, compared to drying with N₂ gas. The method showed excellent linearity with coefficient of variation (R²) > 0.99 and a good precision (relative standard deviation < 20 %) and accuracy. Finally, systemic concentrations of SCFA in human plasma samples could successfully be determined.

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优化和验证分析人体血浆或血清中乙酸盐、丙酸盐和丁酸盐浓度的气相色谱-质谱法

发酵产生的短链脂肪酸（SCFA）4 是膳食纤维摄入对健康有益的潜在媒介。SCFA 通过全身循环影响肠道局部和远处器官的生理过程。由于 SCFA 被用作结肠细胞的能量来源和肝脏代谢的底物，因此其在全身循环中的浓度很低。因此，全身 SCFA 的定量需要灵敏的分析技术。本文介绍了一种气相色谱-质谱分析方法的优化和验证，该方法采用 2,4-二氟苯胺 (DFA)5 进行衍生，并在乙酸乙酯中萃取，用于测量全身 SCFA 的浓度。通过改变 DFA、偶联剂 1,3-二环己基碳二亚胺、乙酸乙酯和用于淬灭衍生化的碳酸氢钠的用量，对样品制备进行了优化。此外，与使用 N2 气体干燥相比，使用真空浓缩器蒸发样品可减少污染，尤其是醋酸盐的污染。该方法线性关系良好，变异系数（R2）为 0.99，精密度（相对标准偏差为 20%）和准确度也很高。最后，该方法成功地测定了人体血浆样本中SCFA的系统浓度。

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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.