Current state of the art of analyte scope in urine metabolome analysis by non-hydrogenative PHIP

Nele Reimets, Kerti Ausmees, Indrek Reile
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Abstract

Non-hydrogenative PHIP (nh-PHIP) is an NMR signal enhancement technique that offers several orders of magnitude gains in detection sensitivity. It is one of the few hyperpolarization methods that have been demonstrated to be applicable to chemical analysis of biological samples and potentially metabolomics. It is, however, a chemoselective method and needs to be tuned to particular analyte and metabolite classes at a time.
Herein, we present a systematic study where we apply four nh-PHIP modifications to urine samples from two different species – human and dog. Firstly, this allows to explore the whole analyte class scope and present what information is nh-PHIP capable of providing by varying the composition of the nh-PHIP catalyst system and the sample preparation protocol. Secondly, comparing hyperpolarized spectra from urines from different species demonstrates that this hyperpolarization technique is robust and tolerant of possibly considerable matrix differences: signals of the same metabolites appear at same chemical shifts from urines that differ from one-another much more than is likely in a realistic metabolomics study. Thereby we propose the idea that nh-PHIP is ready for application in metabolomics experiments.

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利用非氢化 PHIP 分析尿液代谢组中分析物范围的技术现状
非氢化 PHIP(nh-PHIP)是一种核磁共振信号增强技术,可将检测灵敏度提高几个数量级。它是少数几种已被证明适用于生物样本化学分析和潜在代谢组学分析的超极化方法之一。不过,它是一种化学选择性方法,需要针对特定的分析物和代谢物类别进行调整。在这里,我们介绍了一项系统性研究,将四种 nh-PHIP 修饰方法应用于人和狗这两种不同物种的尿液样本。首先,通过改变 nh-PHIP 催化剂系统的组成和样品制备方案,我们可以探索整个分析物类别的范围,并展示 nh-PHIP 能够提供哪些信息。其次,比较不同物种尿液的超极化光谱表明,这种超极化技术具有很强的鲁棒性,并能容忍可能存在的巨大基质差异:尿液中相同代谢物的信号出现在相同的化学位移上,而这些尿液之间的差异要比实际代谢组学研究中可能出现的差异大得多。因此,我们认为 nh-PHIP 可以应用于代谢组学实验。
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