Metabolite quantification by NMR and LC-MS/MS reveals differences between unstimulated, stimulated, and pure parotid saliva
Metabolite quantification by NMR and LC-MS/MS reveals differences between unstimulated, stimulated, and pure parotid saliva
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DOI：
10.1016/j.jpba.2017.03.037
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发表时间：
2017-06-05
影响因子：
3.4
通讯作者：
Ohman, Anders
中科院分区：
文献类型：
--
作者：
Figueira, Joao;Gouveia-Figueira, Sandra;Ohman, Anders
唾液是一种容易获得的生物流体，其对代谢变化敏感，并且可以通过快速和非侵入性的收集程序来收集，并且其显示出用于临床代谢组学研究的巨大前景。这项工作研究了代谢物的组成，和之间的差异，唾液样本收集的无刺激的吐痰/流口水，石蜡咀嚼刺激吐痰，腮腺抽吸使用有针对性的核磁共振（NMR）光谱和液相色谱耦合串联质谱（LC-MS/MS）代谢物定量。正如本文所应用的，这两种分析技术提供了互补的代谢物信息，并通过对可溶性代谢物的稳健NMR定量和对特定代谢途径中的生物活性脂质的灵敏靶向LC-MS/MS分析，共同扩展了代谢物组的覆盖范围。对超滤（3 kDa截止值）唾液样品进行NMR分析，共得到45种定量代谢物。对过滤和未过滤样品进行LC-MS/MS分析，并对两种内源性大麻素（AEA和PEA）和22种氧化脂质进行定量，这是目前对人类唾液中生物活性脂质最全面的靶向分析。在三种唾液样本采集方法之间观察到代谢物组成的重要差异，在设计唾液代谢组学研究时应考虑到这一点。此外，结合使用两个代谢组学平台（NMR和LC-MS/MS）被证明是可行的唾液代谢组的研究和临床研究。(C)2017 Elsevier B. V.版权所有。
Saliva is a readily available biofluid that is sensitive to metabolic changes and can be collected through rapid and non-invasive collection procedures, and it shows great promise for clinical metabolomic studies. This work studied the metabolite composition of, and the differences between, saliva samples collected by unstimulated spitting/drooling, paraffin chewing-stimulated spitting, and parotid gland suction using targeted nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for metabolite quantification. As applied here, these two analytical techniques provide complementary metabolite information and together extend the metabolome coverage with robust NMR quantification of soluble metabolites and sensitive targeted LC-MS/MS analysis of bioactive lipids in specific metabolic pathways. The NMR analysis was performed on ultrafiltrated (3 kDa cutoff) saliva samples and resulted in a total of 45 quantified metabolites. The LC-MS/MS analysis was performed on both filtered and unfiltered samples and resulted in the quantification of two endocannabinoids (AEA and PEA) and 22 oxylipins, which at present is the most comprehensive targeted analysis of bioactive lipids in human saliva. Important differences in the metabolite composition were observed between the three saliva sample collection methods, which should be taken into consideration when designing metabolomic studies of saliva. Furthermore, the combined use of the two metabolomics platforms (NMR and LC-MS/MS) proved to be viable for research and clinical studies of the salivary metabolome. (C) 2017 Elsevier B.V. All rights reserved.