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Pre-analytic evaluation of volumetric absorptive microsampling and integration in a mass spectrometry-based metabolomics workflow

机译：基于质谱的代谢组学工作流程中的体积吸收微采样和积分的预分析评估

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Volumetric absorptive microsampling (VAMS) is a novel approach that allows single-drop (10 μL) blood collection. Integration of VAMS with mass spectrometry (MS)-based untargeted metabolomics is an attractive solution for both human and animal studies. However, to boost the use of VAMS in metabolomics, key pre-analytical questions need to be addressed. Therefore, in this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. We first evaluated the best extraction procedure for the polar metabolome and found that the highest number and amount of metabolites were recovered upon extraction with acetonitrile/water (70:30). In contrast, basic conditions (pH 9) resulted in divergent metabolite profiles mainly resulting from the extraction of intracellular metabolites originating from red blood cells. In addition, the prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but once the VAMS devices were stored at − 80 °C, the metabolome remained stable for up to 6 months. The time used for drying the sample did also affect the metabolome. In fact, some metabolites were rapidly degraded or accumulated in the sample during the first 48 h at room temperature, indicating that a longer drying step will significantly change the concentration in the sample.
机译：容积吸收微量采样（VAMS）是一种允许单滴（10μL）血液采集的新颖方法。 VAMS与基于质谱（MS）的非靶向代谢组学的集成，对于人类和动物研究都是有吸引力的解决方案。但是，为了促进在代谢组学中使用VAMS，需要解决关键的分析前问题。因此，在这项工作中，我们将VAMS整合到了基于MS的非靶向代谢组学工作流程中，并研究了分析前的策略，例如样品提取步骤和在不同储存条件下的代谢组稳定性。我们首先评估了极性代谢物的最佳提取程序，发现用乙腈/水（70:30）提取后，回收的代谢物数量和数量最高。相反，基本条件（pH 9）导致代谢产物分布不同，这主要是由于提取自红细胞的细胞内代谢产物所致。此外，室温下长时间的血液样本存储会导致代谢组的组成发生重大变化，但是一旦VAMS设备存储在− 80°C，代谢组将保持稳定长达6个月。用于干燥样品的时间也确实影响了代谢组。实际上，在室温下的最初48小时内，一些代谢产物在样品中迅速降解或积累，这表明更长的干燥步骤将显着改变样品中的浓度。

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