Quantitative analysis of blood plasma metabolites using isotope enhanced NMR methods.
Quantitative analysis of blood plasma metabolites using isotope enhanced NMR methods.
复制标题
DOI：
10.1021/ac101938w
复制
发表时间：
2010-11-01
影响因子：
7.4
通讯作者：
Raftery, Daniel
中科院分区：
文献类型：
--
作者：
Gowda, G. A. Nagana;Tayyari, Fariba;Ye, Tao;Suryani, Yuliana;Wei, Siwei;Shanaiah, Narasimhamurthy;Raftery, Daniel
核磁共振波谱是一种强有力的定性和定量分析工具。然而，在复杂流体如人血浆中的精确定量分析是具有挑战性的，并且使用一维NMR的分析受到信号重叠的限制。由于灵敏度低，因此在常规基础上使用涉及天然丰度13 C的异谱实验是不切实际的，尽管它们的分辨率有所提高。围绕规避这样的瓶颈，这项研究表明，利用同位素标记的NMR实验的组合来分析人体血浆中的代谢物。同位素标记样品的1H-15 N HSQC和1H-13 C HSQC实验与常规1H一维和1H-1H TOCSY实验相结合，提供了血浆中大量代谢物的定量信息。首先对人血液中常见的28种合成代谢物类似物的混合物进行了测试;然后鉴定并定量了标准NIST（美国国家标准与技术研究所）人血浆中的27种代谢物，其中17种代谢物的平均变异系数为2.4%，考虑所有代谢物时为5.6%。羧酸和胺代表体液中的大部分代谢物，并且通过同位素标记对它们的分析能够显著增强用于生物标志物发现应用的代谢库。通过15 N和13 C同位素标记赋予的NMR实验的改进的灵敏度和分辨率对于可检测的代谢池的增强和血浆代谢物的准确分析都是有吸引力的。该方法可以很容易地扩展到几乎任何生物混合物中的许多其他代谢物。
NMR spectroscopy is a powerful analytical tool for both qualitative and quantitative analysis. However, accurate quantitative analysis in complex fluids such as human blood plasma is challenging, and analysis using one-dimensional NMR is limited by signal overlap. It is impractical to use heteronuclear experiments involving natural abundance 13C on a routine basis due to low sensitivity, despite their improved resolution. Focusing on circumventing such bottlenecks, this study demonstrates the utility of a combination of isotope tagged NMR experiments to analyze metabolites in human blood plasma. 1H-15N HSQC and 1H-13C HSQC experiments on the isotope tagged samples combined with the conventional 1H one-dimensional and 1H-1H TOCSY experiments provide quantitative information on a large number of metabolites in plasma. The methods were first tested on a mixture of 28 synthetic analogues of metabolites commonly present in human blood; twenty-seven metabolites in a standard NIST (National Institute of Standards and Technology) human blood plasma were then identified and quantified with an average coefficient of variation of 2.4 % for 17 metabolites and 5.6% when all the metabolites were considered. Carboxylic acids and amines represent a majority of the metabolites in body fluids and their analysis by isotope tagging enables a significant enhancement of the metabolic pool for biomarker discovery applications. Improved sensitivity and resolution of NMR experiments imparted by 15N and 13C isotope tagging is attractive for both the enhancement of the detectable metabolic pool and accurate analysis of plasma metabolites. The approach can be easily extended to many additional metabolites in almost any biological mixture.
登录
查看更多内容
HMDB：人类代谢组数据库。
影响因子：
14.9
作者：
Wishart, David S;Tzur, Dan;Knox, Craig;Eisner, Roman;Guo, An Chi;Young, Nelson;Cheng, Dean;Jewell, Kevin;Arndt, David;Sawhney, Summit;Fung, Chris;Nikolai, Lisa;Lewis, Mike;Coutouly, Marie-Aude;Forsythe, Ian;Tang, Peter;Shrivastava, Savita;Jeroncic, Kevin;Stothard, Paul;Amegbey, Godwin;Block, David;Hau, David D;Wagner, James;Miniaci, Jessica;Clements, Melisa;Gebremedhin, Mulu;Guo, Natalie;Zhang, Ying;Duggan, Gavin E;Macinnis, Glen D;Weljie, Alim M;Dowlatabadi, Reza;Bamforth, Fiona;Clive, Derrick;Greiner, Russ;Li, Liang;Marrie, Tom;Sykes, Brian D;Vogel, Hans J;Querengesser, Lori
通讯作者：
Querengesser, Lori
影响因子：
7.4
作者：
de Graaf, RA;Behar, KL
通讯作者：
Behar, KL
影响因子：
7.4
作者：
Rai, Ratan Kumar;Tripathi, Pratima;Sinha, Neeraj
通讯作者：
Sinha, Neeraj
影响因子：
1.8
作者：
Nicholson, JK;Lindon, JC;Holmes, E
通讯作者：
Holmes, E
影响因子：
7.4
作者：
Fonville, Judith M.;Maher, Anthony D.;Nicholson, Jeremy K.
通讯作者：
Nicholson, Jeremy K.