脂质组学和代谢组学的新兴学科显示出发现诊断生物标志物的巨大潜力,但适当的分析前样品处理程序是关键的,因为在样品收集过程中,几种分析物易于离体变形。为了测试来自K3EDTA全血收集管的血浆样品的中间储存温度和储存期如何影响分析物浓度,我们评估了非空腹健康志愿者(n=9)的广谱代谢物样本,包括脂质和脂质介质,使用完善的基于LC-MS的平台。我们使用基于倍数变化的方法作为分析物稳定性的相对量度来评估489种分析物,采用靶向LC-MS/MS和LC-HRMS筛查的组合。许多分析物的浓度被发现是可靠的,通常证明不太严格的样品处理是合理的;然而,某些分析物不稳定,配套需要细致的加工。我们为严格程度不同的样品处理方案提出了四个数据驱动的建议,基于分析物的最大数量和常规临床实施的可行性。这些方案还能够基于其对离体畸变的分析物特异性脆弱性来简单评估生物标志物候选物。总之,分析前样品处理对某些代谢物作为生物标志物的适用性有重大影响,包括几种脂质和脂质介质。我们的样品处理建议将提高样品的可靠性和质量,当这些代谢物是常规临床诊断所必需时。
The emerging disciplines of lipidomics and metabolomics show great potential for the discovery of diagnostic biomarkers, but appropriate pre-analytical sample-handling procedures are critical because several analytes are prone to ex vivo distortions during sample collection. To test how the intermediate storage temperature and storage period of plasma samples from K3EDTA whole-blood collection tubes affect analyte concentrations, we assessed samples from non-fasting healthy volunteers (n = 9) for a broad spectrum of metabolites, including lipids and lipid mediators, using a well-established LC-MS-based platform. We used a fold change-based approach as a relative measure of analyte stability to evaluate 489 analytes, employing a combination of targeted LC-MS/MS and LC-HRMS screening. The concentrations of many analytes were found to be reliable, often justifying less strict sample handling; however, certain analytes were unstable, supporting the need for meticulous processing. We make four data-driven recommendations for sample-handling protocols with varying degrees of stringency, based on the maximum number of analytes and the feasibility of routine clinical implementation. These protocols also enable the simple evaluation of biomarker candidates based on their analyte-specific vulnerability to ex vivo distortions. In summary, pre-analytical sample handling has a major effect on the suitability of certain metabolites as biomarkers, including several lipids and lipid mediators. Our sample-handling recommendations will increase the reliability and quality of samples when such metabolites are necessary for routine clinical diagnosis.