基于NMR的内分子13C分布自然丰富的方法适应小量的葡萄糖
Sophie Renou1, Mathilde Grand1, Valérie Daux2
1CEISAM, CNRS, Nantes Université, F-44322 Nantes, France.
Analytical chemistry
|July 6, 2023
概括
使用新的2D-NMR序列的定量核磁共振 (NMR) 能够对小型生物样本进行精确的,位置特定的碳同位素分析 (δ13C). 这一进步大大减少了对同位素比率测量的材料要求和实验时间.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 频谱学是一种光谱学.
背景情况:
- 量化核磁共振 (NMR) 用于同位素比测量 (irm-NMR) 对于位置特定的碳同位素分析 (δ13C) 是有价值的.
- 以前的IRM-NMR方法使用单脉冲序列,需要大量的样本和长时间的实验,限制了在生物组织中的应用.
- 研究植物中的糖代谢已经利用irm-NMR对衍生葡萄糖进行了研究.
研究的目的:
- 适应和优化2D-NMR序列用于定量同位素比率测量.
- 为了能够高精度 (<1 mUr) 分析小样本量 (10 mg) 的位置特定的 δ13C.
- 开发一种方法,以对原始2D-NMR数据进行校正,以达到标准的13C尺度.
主要方法:
- 研究了2D-NMR分析用于定量同位素比率测量的使用.
- 优化了2D-NMR序列,用于分析少量 (10毫克) 的葡萄糖衍生物 (二甲胺葡萄糖,DAGF).
- 开发了一种使用参考材料和用单脉冲序列进行比较分析的数据校正方法.
主要成果:
- 通过使用优化的2D-NMR序列,在每个C原子位置成功分析了少量 (10毫克) DAGF,精度高于1mUr.
- 建立了一种校正方法,以在传统的 δ 13 C 尺度上表达原始 C 丰度.
- 使用单脉冲和2D-NMR序列,比较来自不同植物代谢 (C3,C4,CAM) 的葡萄糖.
结论:
- 开发的2D-NMR方法显著减少了对特定位置同位素比率测量的样本要求和分析时间.
- 该方法提供了高精度和准确性,适用于分析生物样本和研究代谢途径.
- 该方法与绿色分析化学原则保持一致,通过尽量减少样品和资源的使用.
相关概念视频
¹³C NMR: ¹H–¹³C Decoupling
1.1K
The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
1.1K
Carbon-13 (¹³C) NMR: Overview
5.8K
Carbon-13 is a naturally occurring NMR-active isotope of carbon with a low natural abundance of 1.1%. In contrast, carbon-12 is the most abundant isotope of carbon with zero nuclear spin. Therefore, it is NMR inactive. The gyromagnetic ratio of carbon-13 is smaller than that of protons. As a result, carbon-13 resonance is about 6000 times weaker than proton resonance. For a given magnetic field strength, the resonance frequency of carbon-13 is about one-fourth of the resonance frequency for...
5.8K
Chemical Shift: Internal References and Solvent Effects
680
In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...
680
Other Nuclides: 31P, 19F, 15N NMR
418
Many organic, inorganic, and biological molecules contain spin-half nuclei such as nitrogen-15, fluorine-19, and phosphorus-31. As a result, NMR studies of these nuclei have found extensive applications in chemical and biological research.
While fluorine-19 and phosphorous-31 have high natural abundances (100%) and positive gyromagnetic ratios, nitrogen-15 has a low natural abundance and a negative gyromagnetic ratio. However, nitrogen-15 is still preferred over nitrogen-14 (which has a...
While fluorine-19 and phosphorous-31 have high natural abundances (100%) and positive gyromagnetic ratios, nitrogen-15 has a low natural abundance and a negative gyromagnetic ratio. However, nitrogen-15 is still preferred over nitrogen-14 (which has a...
418
Applications Of NMR In Biology
3.7K
Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
3.7K
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)
1.1K
When proton-coupled carbon-13 spectra are simplified by a broadband proton decoupling technique, structural information about the coupled protons is lost. Distortionless enhancement by polarization transfer (DEPT) is a technique that provides information on the number of hydrogens attached to each carbon in a molecule. While the DEPT experiment utilizes complex pulse sequences, the pulse delay and flip angle are specifically manipulated. The resulting signals have different phases depending on...
1.1K


