Quantification of high-resolution ¹H-[¹³C] NMR spectra from rat brain extracts.
Quantification of high-resolution ¹H-[¹³C] NMR spectra from rat brain extracts.
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DOI：
10.1021/ac5006926
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发表时间：
2014-05-20
影响因子：
7.4
通讯作者：
Behar, Kevin L.
中科院分区：
文献类型：
--
作者：
de Graaf, Robin A.;Chowdhury, Golam M. I.;Behar, Kevin L.
核磁共振波谱结合13 C标记的底物输注是一种独特的技术，以获得有关动态代谢通量的信息，在体内无创。在许多情况下，在啮齿动物动态13 C研究过程中获得的体内信息内容可以通过对脑提取物进行高分辨率1H-[13 C] NMR光谱学来增强。此前，已有研究表明，利用化学位移和标量耦合的完整先验知识，利用模拟基组，通过光谱拟合程序可以准确量化大鼠脑提取物的1H NMR光谱。将13 C标记引入到各种代谢物中呈现出需要修改光谱拟合例程的复杂性。由于给定分子内的不同多重峰积累了不同量的13 C标记，因此必须放弃1H NMR谱中典型的多重峰之间的固定振幅关系。此外，13 C同位素效应导致光谱多重峰模式，其变得依赖于13 C标记累积的量，从而防止使用共同的基组。这里提出了一个修改后的光谱拟合例程，可容纳可变的13 C标记积累和13 C同位素效应。将光谱拟合结果与色谱柱分离样品的手动积分进行定量比较，其中光谱重叠最小化。
NMR spectroscopy in combination with 13C-labeled substrate infusion is a unique technique to obtain information about dynamic metabolic fluxes noninvasively in vivo. In many cases, the in vivo information content obtained during dynamic 13C studies in rodents can be enhanced by high-resolution 1H-[13C] NMR spectroscopy on brain extracts. Previously, it has been shown that 1H NMR spectra from rat brain extracts can be accurately quantified with a spectral fitting routine utilizing simulated basis sets using complete prior knowledge of chemical shifts and scalar couplings. The introduction of 13C label into the various metabolites presents complications that demand modifications of the spectral fitting routine. As different multiplets within a given molecule accumulate various amounts of 13C label, the fixed amplitude relationship between multiplets typical for 1H NMR spectra must be abandoned. In addition, 13C isotope effects lead to spectral multiplet patterns that become dependent on the amount of 13C label accumulation, thereby preventing the use of a common basis set. Here a modified spectral fitting routine is presented that accommodates variable 13C label accumulation and 13C isotope effects. Spectral fitting results are quantitatively compared to manual integration on column-separated samples in which spectral overlap is minimized.
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影响因子：
7.4
作者：
de Graaf, Robin A.;Chowdhury, Golam M. I.;Behar, Kevin L.
通讯作者：
Behar, Kevin L.
影响因子：
2.2
作者：
Mlynarik, Vladimir;Cudalbu, Cristina;Gruetter, Rolf
通讯作者：
Gruetter, Rolf
影响因子：
2.2
作者：
Pfeuffer, J;Tkác, I;Gruetter, R
通讯作者：
Gruetter, R
DOI：
10.1073/pnas.0405065101
发表时间：
2004-08-24
影响因子：
11.1
作者：
de Graaf, RA;Mason, GF;Behar, KL
通讯作者：
Behar, KL
影响因子：
2.9
作者：
Henry, PG;Öz, G;Gruetter, R
通讯作者：
Gruetter, R