Straube Henryk, Witte Claus-Peter, Herde Marco
Department of Molecular Nutrition and Biochemistry of Plants, Leibniz Universität Hannover, 30419 Hannover, Germany.
Cells. 2021 Mar 20;10(3):689. doi: 10.3390/cells10030689.
Nucleotides fulfill many essential functions in plants. Compared to non-plant systems, these hydrophilic metabolites have not been adequately investigated in plants, especially the less abundant nucleotide species such as deoxyribonucleotides and modified or damaged nucleotides. Until recently, this was mainly due to a lack of adequate methods for in-depth analysis of nucleotides and nucleosides in plants. In this review, we focus on the current state-of-the-art of nucleotide analysis in plants with liquid chromatography coupled to mass spectrometry and describe recent major advances. Tissue disruption, quenching, liquid-liquid and solid-phase extraction, chromatographic strategies, and peculiarities of nucleotides and nucleosides in mass spectrometry are covered. We describe how the different steps of the analytical workflow influence each other, highlight the specific challenges of nucleotide analysis, and outline promising future developments. The metabolite matrix of plants is particularly complex. Therefore, it is likely that nucleotide analysis methods that work for plants can be applied to other organisms as well. Although this review focuses on plants, we also discuss advances in nucleotide analysis from non-plant systems to provide an overview of the analytical techniques available for this challenging class of metabolites.
核苷酸在植物中发挥着许多重要功能。与非植物系统相比，这些亲水性代谢物在植物中尚未得到充分研究，尤其是脱氧核糖核苷酸以及修饰或受损的核苷酸等含量较少的核苷酸种类。直到最近，这主要是由于缺乏对植物中核苷酸和核苷进行深入分析的适当方法。在这篇综述中，我们重点介绍了液相色谱-质谱联用技术在植物核苷酸分析方面的最新进展，并描述了近期的主要突破。内容涵盖了组织破碎、淬灭、液-液萃取和固相萃取、色谱策略以及核苷酸和核苷在质谱分析中的特性。我们阐述了分析流程的不同步骤是如何相互影响的，突出了核苷酸分析的具体挑战，并概述了有前景的未来发展方向。植物的代谢物基质尤为复杂。因此，适用于植物的核苷酸分析方法很可能也可应用于其他生物。尽管本综述聚焦于植物，但我们也讨论了非植物系统中核苷酸分析的进展，以便对这类具有挑战性的代谢物的可用分析技术进行概述。
