Dunn Warwick B, Broadhurst David, Ellis David I, Brown Marie, Halsall Anthony, O'Hagan Steven, Spasic Irena, Tseng Andrew, Kell Douglas B
Bioanalytical Sciences Group, School of Chemistry, Manchester Interdiscplinary Biocentre, University of Manchester, 131 Princess Street, Manchester, M1 7ND, UK.
Int J Epidemiol. 2008 Apr;37 Suppl 1:i23-30. doi: 10.1093/ije/dym281.
The stability of mammalian serum and urine in large metabolomic investigations is essential for accurate, valid and reproducible studies. The stability of mammalian serum and urine, either processed immediately by freezing at -80 degrees C or stored at 4 degrees C for 24 h before being frozen, was compared in a pilot metabolomic study of samples from 40 separate healthy volunteers.
Metabolic profiling with GC-TOF-MS was performed for serum and urine samples collected from 40 volunteers and stored at -80 degrees C or 4 degrees C for 24 h before being frozen at -80 degrees C. Subsequent Wilcoxon rank sum test and Principal Components Analysis (PCA) methods were used to assess whether differences in the metabolomes were detected between samples stored at 4 degrees C for 0 or 24 h.
More than 700 unique metabolite peaks were detected, with over 200 metabolite peaks detected in any one sample. PCA and Wilcoxon rank sum tests of serum and urine data showed as a general observation that the variance associated with the replicate analysis per sample (analytical variance) was of the same magnitude as the variance observed between samples stored at 4 degrees C for 0 or 24 h. From a functional point of view the metabolomic composition of the majority of samples did not change in a statistically significant manner when stored under two different conditions.
Based on this small pilot study, the UK Biobank sampling, transport and fractionation protocols are considered suitable to provide samples, which can produce scientifically robust and valid data in metabolomic studies.
在大规模代谢组学研究中，哺乳动物血清和尿液的稳定性对于准确、有效和可重复的研究至关重要。在一项针对40名独立健康志愿者样本的初步代谢组学研究中，比较了哺乳动物血清和尿液的稳定性，这些样本要么立即在-80℃冷冻处理，要么在4℃储存24小时后再冷冻。
对从40名志愿者收集的血清和尿液样本进行气相色谱-飞行时间质谱（GC-TOF-MS）代谢谱分析，样本在-80℃冷冻前在4℃储存0或24小时。随后使用Wilcoxon秩和检验和主成分分析（PCA）方法评估在4℃储存0或24小时的样本之间是否检测到代谢组的差异。
检测到700多个独特的代谢物峰，任何一个样本中检测到的代谢物峰超过200个。血清和尿液数据的PCA和Wilcoxon秩和检验总体表明，每个样本重复分析相关的方差（分析方差）与在4℃储存0或24小时的样本之间观察到的方差大小相同。从功能角度来看，大多数样本在两种不同条件下储存时，其代谢组组成没有发生统计学上的显著变化。
基于这项小型初步研究，英国生物银行的采样、运输和分馏方案被认为适合提供样本，这些样本能够在代谢组学研究中产生科学可靠和有效的数据。
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