A critical evaluation of the use of gas chromatography- and high performance liquid chromatography-mass spectrometry techniques for the analysis of microbial metabolites in human urine after consumption of orange juice
A critical evaluation of the use of gas chromatography- and high performance liquid chromatography-mass spectrometry techniques for the analysis of microbial metabolites in human urine after consumption of orange juice
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DOI：
10.1016/j.chroma.2018.09.016
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发表时间：
2018-11-09
影响因子：
4.1
通讯作者：
Manuel Moreno-Rojas, Jose
中科院分区：
文献类型：
--
作者：
Luis Ordonez, Jose;Pereira-Caro, Gema;Manuel Moreno-Rojas, Jose
本研究比较并验证了两种分析方法（HPLC-HRMS 和 GC-MS），使用 MSTFA 作为衍生剂，用于分析尿液中累积的微生物衍生的酚酸和芳香族化合物，这些物质是在饮用 500 mL 橙汁后 24 小时内收集的。此外，当使用 HPLC-HRMS 技术时，比较了使用 SDB-L 和 HLB 固相柱的纯化程序。 HPLC-HRMS 和 GC-MS 方法在特异性、灵敏度、检测限和定量、回收率和基质效应方面均得到成功验证。与 GC-MS 不同，HPLC-HRMS 不需要在分析前对样品进行衍生化。由于缺乏挥发性，GC-MS 不适用于硫酸酚和葡萄糖醛酸代谢物的分析。然而，这些 II 相代谢物可以通过 HPLC-HRMS 进行分析，从而提供有关源自微生物介导的橙汁（多）酚降解的酚类化合物的更详细和完整的信息。此外，在 HPLC-HRMS 分析之前使用 SDB-L 和 HLB 柱进行样品纯化适用于游离酚类和葡萄糖醛酸代谢物，但不适用于硫酸盐衍生物。这些发现强调，使用不适当的分析方案可能会对膳食（多）酚的生物利用度研究产生不利影响，其中微生物群衍生的酚类分解代谢物发挥着重要作用。 (C) 2018 Elsevier B.V. 保留所有权利。
The present study compared and validated two analytical methods, HPLC-HRMS, and GC-MS using MSTFA as derivatization agent, for the analysis of microbiota-derived phenolic acids and aromatic compounds accumulating in urine, collected over a 24 h period after the consumption of 500 mL of orange juice. In addition, purification procedures using SDB-L and HLB solid phase cartridges were compared when HPLC- HRMS technique was used. Both HPLC-HRMS and GC-MS methodologies were successfully validated in terms of specificity, sensitivity, limit of detection and quantification, recovery and matrix effects. HPLC- HRMS, unlike GC-MS, does not require sample derivatization prior to analysis. GC-MS was not suitable for the analysis of phenolic sulfate and glucuronide metabolites because of their lack of volatility. These phase II metabolites could, however, be analysed by HPLC-HRMS which, as a consequence, provided more detailed and complete information on the phenolic compounds derived from microbiota-mediated degradation of orange juice (poly)phenols. Furthermore, the use of SDB-L and HLB cartridges for sample purification prior to HPLC-HRMS analysis is suitable for free phenolics and glucuronide metabolites but not sulfate derivatives. These findings highlight that the use of an inappropriate analytical protocol can adversely affect studies on the bioavailability of dietary (poly)phenols in which microbiota-derived phenolic catabolites play an important role. (C) 2018 Elsevier B.V. All rights reserved.