A key limiting step for high-throughput NMR-based metabolomics is the lack of rapid and accurate tools for absolute quantification of many metabolites. We developed, implemented, and evaluated an algorithm, AQuA (Automated Quantification Algorithm), for targeted metabolite quantification from complex ¹H NMR spectra. AQuA operates based on spectral data extracted from a library consisting of one standard calibration spectrum for each metabolite. It uses one preselected NMR signal per metabolite for determining absolute concentrations and does so by effectively accounting for interferences caused by other metabolites. AQuA was implemented and evaluated using experimental NMR spectra from human plasma. The accuracy of AQuA was tested and confirmed in comparison with a manual spectral fitting approach using the ChenomX software, in which 61 out of 67 metabolites quantified in 30 human plasma spectra showed a goodness-of-fit (r²) close to or exceeding 0.9 between the two approaches. In addition, three quality indicators generated by AQuA, namely, occurrence, interference, and positional deviation, were studied. These quality indicators permit evaluation of the results each time the algorithm is operated. The efficiency was tested and confirmed by implementing AQuA for quantification of 67 metabolites in a large data set comprising 1342 experimental spectra from human plasma, in which the whole computation took less than 1 s.

中文翻译：

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AQuA：基于高通量核磁共振的代谢组学的自动定量算法及其在人血浆中的应用

高通量基于NMR的代谢组学的关键限制步骤是缺乏快速准确的工具，无法对多种代谢物进行绝对定量。我们开发，实施和评估了一种算法AQuA（自动定量算法），用于复杂^1的目标代谢物定量1 H NMR谱。AQuA基于从库中提取的光谱数据进行操作，该库由每种代谢物的一个标准校准光谱组成。它使用每个代谢物一个预选的NMR信号来确定绝对浓度，并通过有效地考虑其他代谢物引起的干扰来确定绝对浓度。使用人血浆的实验NMR光谱实施并评估了AQuA。通过使用ChenomX软件与手动光谱拟合方法进行比较，测试并证实了AQuA的准确性，其中在30个人血浆中量化的67种代谢产物中，有61种的拟合度（r ²）接近或超过0.9两种方法之间。此外，AQuA产生了三个质量指标，即发生率，研究了干涉和位置偏差。这些质量指标允许每次算法运行时对结果进行评估。通过实施AQuA定量分析大型数据集（包括来自人类血浆的1342个实验光谱）中的67种代谢物的效率，进行了测试并证实了其效率，整个过程花费了不到1 s的时间。