The influence of EDTA and citrate anticoagulant addition to human plasma on information recovery from NMR-based metabolic profiling studies
The influence of EDTA and citrate anticoagulant addition to human plasma on information recovery from NMR-based metabolic profiling studies
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DOI：
10.1039/b907021d
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发表时间：
2010-01-01
影响因子：
--
通讯作者：
Lindon, John C.
中科院分区：
文献类型：
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作者：
Barton, Richard H.;Waterman, Daniel;Lindon, John C.
广泛使用的血液抗凝剂柠檬酸盐和EDTA在流行病学和临床研究中收集的血浆样本的H-1 NMR光谱中产生突出峰，这些对恢复内源性代谢物的生化信息造成不同程度的干扰。为了研究这些物质引起的潜在代谢信息丢失以及抗凝剂和内源性组分之间任何可能的分子间相互作用，分析了从20名个体中采集的40份柠檬酸盐或EDTA分离人血浆样本的H-1 NMR光谱。内源性代谢物峰被较大的柠檬酸盐峰或游离EDTA及其钙和镁复合物峰选择性遮蔽。结果表明，产生被EDTA或柠檬酸盐遮蔽的峰的内源性代谢物几乎总是具有其他共振，从而可以对其进行鉴别和潜在定量。此外，代谢信息恢复可以最大限度地通过使用频谱编辑技术，如自旋回波，扩散编辑和J-分辨实验。发现添加的柠檬酸盐或EDTA与内源性组分之间的任何相互作用的NMR光谱效应可忽略不计。最后，使用简单的多元统计方法，如主成分分析，分离样本的识别是可行的。因此，即使使用NMR不合适的柠檬酸盐或EDTA抗凝剂收集了遗留流行病学血浆样本，仍然可以有效地回收有用的生化信息。
The widely-used blood anticoagulants citrate and EDTA give rise to prominent peaks in H-1 NMR spectra of plasma samples collected in epidemiological and clinical studies, and these cause varying levels of interference in recovering biochemical information on endogenous metabolites. To investigate both the potential metabolic information loss caused by these substances and any possible inter-molecular interactions between the anticoagulants and endogenous components, the H-1 NMR spectra of 40 split human plasma samples collected from 20 individuals into either citrate or EDTA have been analysed. Endogenous metabolite peaks were selectively obscured by large citrate peaks or those from free EDTA and its calcium and magnesium complexes. It is shown that the endogenous metabolites that give rise to peaks obscured by those from EDTA or citrate almost invariably also have other resonances that allow their identification and potential quantitation. Also, metabolic information recovery could be maximised by use of spectral editing techniques such as spin-echo, diffusion-editing and J-resolved experiments. The NMR spectral effects of any interactions between the added citrate or EDTA and endogenous components were found to be negligible. Finally, identification of split samples was feasible using simple multivariate statistical approaches such as principal components analysis. Thus even when legacy epidemiological plasma samples have been collected using the NMR-inappropriate citrate or EDTA anticoagulants, useful biochemical information can still be recovered effectively.