Lipid extraction from dried blood spots and dried milk spots for untargeted high throughput lipidomics.,Molecular Omics

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Lipid extraction from dried blood spots and dried milk spots for untargeted high throughput lipidomics.
Molecular Omics ( IF 3.0 ) Pub Date : 2020-09-10 , DOI: 10.1039/d0mo00102c
Samuel Furse ₁ , Albert Koulman

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Dried blood spots (DBS) and dried milk spots (DMS) represent convenient matrices for collecting and storing human samples. However, the use of these sample types for researching lipid metabolism remains relatively poorly explored, and especially unclear is the efficiency of lipid extraction in the context of high throughput, untargeted lipidomics. A visual inspection of punched DBSs after standard extraction suggests that the samples remain largely intact. DMSs comprise a dense aggregate of milk fat globules on one side of the card, suggesting that part of the lipid fraction may be physically inaccessible. This led us to the hypothesis that decoagulating may facilitate lipid extraction from both DBSs and DMSs. We tested this hypothesis using a mixture of strong chaeotropes (guanidine and thiourea) in both DBS and DMS in the context of high throughput lipidomics (96/384w plate). Extraction of lipids from DMSs was tested with established extractions and one novel solvent mixture in a high throughput format. We found that exposure of DBSs to chaeotropes facilitated collection of the lipid fraction but was ineffective for DMSs. The lipid fraction of DMSs was best isolated without water, using a mixture of xylene/methanol/isopropanol (1 : 2 : 4). We conclude that decoagulation is essential for efficient extraction of lipids from DBSs and that a non-aqueous procedure using a spectrum of solvents is the best procedure for extracting lipids from DMSs. These methods represent convenient steps that are compatible with the sample structure and type, and with high throughput lipidomics.

中文翻译：

从干血斑和干奶斑中提取脂质，用于非靶向高通量脂质组学。

干血斑 (DBS) 和干奶斑 (DMS) 代表了用于收集和储存人体样本的便捷基质。然而，使用这些样本类型来研究脂质代谢的研究仍然相对较少，尤其是在高通量、非靶向脂质组学的背景下脂质提取的效率还不清楚。标准提取后对冲孔 DBS 的目视检查表明样品基本保持完好。DMS 在卡片的一侧包含致密的乳脂球聚集体，这表明部分脂质部分可能在物理上无法接近。这导致我们假设去凝可能有助于从 DBS 和 DMS 中提取脂质。我们在高通量脂质组学（96/384w 板）的背景下，在 DBS 和 DMS 中使用强离液剂（胍和硫脲）的混合物测试了这一假设。从 DMS 中提取脂质已通过已建立的提取物和一种高通量形式的新型溶剂混合物进行测试。我们发现将 DBS 暴露于离液素促进了脂质部分的收集，但对 DMS 无效。使用二甲苯/甲醇/异丙醇 (1 : 2 : 4) 的混合物，最好在不加水的情况下分离 DMS 的脂质部分。我们得出结论，去凝对于从 DBS 中有效提取脂质至关重要，并且使用一系列溶剂的非水性程序是从 DMS 中提取脂质的最佳程序。这些方法代表了与样品结构和类型兼容的便捷步骤，

更新日期：2020-11-03

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