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Evaluation of Fumaric Acid and Maleic Acid as Internal Standards for NMR Analysis of Protein Precipitated Plasma, Serum, and Whole Blood
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-02-04 , DOI: 10.1021/acs.analchem.0c04766 G A Nagana Gowda , Natalie N Hong , Daniel Raftery 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-02-04 , DOI: 10.1021/acs.analchem.0c04766 G A Nagana Gowda , Natalie N Hong , Daniel Raftery 1
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Significant advances have been made in unknown metabolite identification and expansion of the number of quantifiable metabolites in human plasma, serum, and whole blood using NMR spectroscopy. However, reliable quantitation of metabolites is still a challenge. A major bottleneck is the lack of a suitable internal standard that does not interact with the complex blood sample matrix and also does not overlap with metabolite peaks apart from exhibiting other favorable characteristics. With the goal of addressing this challenge, a comprehensive investigation of fumaric and maleic acids as potential internal standards was made along with a comparison with the conventional standards, TSP (trimethylsilylpropionic acid) and DSS (trimethylsilylpropanesulfonic acid). Both fumaric acid and maleic acid exhibited a surprisingly high performance with a quantitation error <1%, while the TSP and DSS caused an average error of up to 35% in plasma, serum, and whole blood. Further, the results indicate that while fumaric acid is a robust standard for all three biospecimens, maleic acid is suitable for only plasma and serum. Maleic acid is not suited for the analysis of whole blood due to its overlap with coenzyme peaks. These findings provide new opportunities for improved and accurate quantitation of metabolites in human plasma, serum, and whole blood using NMR spectroscopy. Moreover, the use of protein precipitation prior to NMR analysis mirrors the sample preparation commonly used for mass spectrometry based metabolomics, such that these findings further strengthen efforts to combine and compare NMR and MS based metabolite data of human plasma, serum, and whole blood for metabolomics based research.
中文翻译：
评价富马酸和马来酸作为蛋白质沉淀血浆、血清和全血 NMR 分析的内标
使用 NMR 光谱在未知代谢物鉴定和扩大人血浆、血清和全血中可量化代谢物数量方面取得了重大进展。然而，代谢物的可靠定量仍然是一个挑战。一个主要瓶颈是缺乏合适的内标，该内标不与复杂的血液样品基质相互作用，也不与代谢物峰重叠，除了表现出其他有利特征。为了应对这一挑战，我们对作为潜在内标的富马酸和马来酸进行了全面研究，并与传统标准 TSP（三甲基甲硅烷基丙酸）和 DSS（三甲基甲硅烷基丙磺酸）进行了比较。富马酸和马来酸都表现出令人惊讶的高性能，定量误差 <1%，而 TSP 和 DSS 在血浆、血清和全血中的平均误差高达 35%。此外，结果表明，虽然富马酸是所有三种生物样本的可靠标准，但马来酸仅适用于血浆和血清。马来酸不适合分析全血，因为它与辅酶峰重叠。这些发现为使用 NMR 光谱法改进和准确定量人血浆、血清和全血中的代谢物提供了新的机会。此外，在 NMR 分析之前使用蛋白质沉淀反映了通常用于基于质谱的代谢组学的样品制备，
更新日期：2021-02-16
中文翻译：
评价富马酸和马来酸作为蛋白质沉淀血浆、血清和全血 NMR 分析的内标
使用 NMR 光谱在未知代谢物鉴定和扩大人血浆、血清和全血中可量化代谢物数量方面取得了重大进展。然而，代谢物的可靠定量仍然是一个挑战。一个主要瓶颈是缺乏合适的内标，该内标不与复杂的血液样品基质相互作用，也不与代谢物峰重叠，除了表现出其他有利特征。为了应对这一挑战，我们对作为潜在内标的富马酸和马来酸进行了全面研究，并与传统标准 TSP（三甲基甲硅烷基丙酸）和 DSS（三甲基甲硅烷基丙磺酸）进行了比较。富马酸和马来酸都表现出令人惊讶的高性能，定量误差 <1%，而 TSP 和 DSS 在血浆、血清和全血中的平均误差高达 35%。此外，结果表明，虽然富马酸是所有三种生物样本的可靠标准，但马来酸仅适用于血浆和血清。马来酸不适合分析全血，因为它与辅酶峰重叠。这些发现为使用 NMR 光谱法改进和准确定量人血浆、血清和全血中的代谢物提供了新的机会。此外，在 NMR 分析之前使用蛋白质沉淀反映了通常用于基于质谱的代谢组学的样品制备，
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