Sheedy John Robert
Department of Zoology, The University of Melbourne, Parkville, VIC, Australia.
Methods Mol Biol. 2013;1055:81-97. doi: 10.1007/978-1-62703-577-4_7.
NMR-based biochemical profiling of natural products has become popular due to the development of high-resolution instruments (>400 MHz) and cryogenically cooled probes/preamplifiers, by increasing the sensitivity of NMR instruments several fold and reducing instrument noise levels. NMR provides a rapid, nondestructive, high-throughput method that requires minimal sample preparation, therefore maintaining the biological integrity of the sample. One-dimensional (1D) solution-state (1)H NMR is used in untargeted sample screening (metabolomics/metabonomics) to gain insight into spectral pattern changes associated with samples of different origins. Metabolomics and metabonomics contextually explains the systematic and quantitative measurement of metabolites that are produced from the biochemical reactions of living systems. This chapter describes some commonly used (1)H NMR experiments for identification and quantification of small molecular weight, water soluble metabolites in biological samples, some considerations for choosing the correct NMR experiment, and sample preparation protocols for isolating metabolites from a number of biological sample types.
由于高分辨率仪器（>400 MHz）以及低温冷却探头/前置放大器的发展，通过将核磁共振（NMR）仪器的灵敏度提高数倍并降低仪器噪声水平，基于NMR的天然产物生化分析已变得流行。NMR提供了一种快速、无损、高通量的方法，所需样品制备极少，因此能保持样品的生物完整性。一维（1D）溶液态氢核磁共振（¹H NMR）用于非靶向样品筛选（代谢组学/代谢物组学），以深入了解与不同来源样品相关的光谱模式变化。代谢组学和代谢物组学从背景角度解释了对生物系统生化反应产生的代谢物进行系统和定量测量的过程。本章介绍了一些常用于鉴定和定量生物样品中小分子水溶性代谢物的¹H NMR实验、选择正确NMR实验的一些注意事项以及从多种生物样品类型中分离代谢物的样品制备方案。
