Measurement of absolute concentrations of individual compounds in metabolite mixtures by gradient-selective time-zero 1H-13C HSQC with two concentration references and fast maximum likelihood reconstruction analysis.
Measurement of absolute concentrations of individual compounds in metabolite mixtures by gradient-selective time-zero 1H-13C HSQC with two concentration references and fast maximum likelihood reconstruction analysis.
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DOI：
10.1021/ac201948f
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发表时间：
2011-12-15
影响因子：
7.4
通讯作者：
Markley, John L.
中科院分区：
文献类型：
--
作者：
Hu, Kaifeng;Ellinger, James J.;Chylla, Roger A.;Markley, John L.
与传统的 2D NMR 相比，零时 2D 13C HSQC (HSQC0) 光谱在定量分析含有化合物混合物的溶液方面具有优势，因为信号强度与成分浓度成正比。 HSQC0 频谱是通过将重复时间外推至零，从基本 HSQC 块内随着重复次数增加而收集的一系列频谱得出的。在这里，我们提出了另一种数据收集方法，即梯度选择性零时间 1H-13C HSQC0 与快速最大似然重建 (FMLR) 数据分析相结合，并使用两个浓度参考来确定绝对浓度。梯度选择性数据采集可产生更清晰的光谱，并且可以在恒定时间和非恒定时间模式下采集 NMR 数据。 FMLR 方法支持半自动数据分析，该方法用于对光谱进行解卷积并提取峰体积。通过参考两种已知浓度的内参化合物的峰体积，将从此分析中获得的峰体积转换为绝对浓度：低浓度限值的 DSS（4,4-二甲基-4-硅戊烷-1-磺酸）（也用作化学位移参考）和高浓度限值的 MES（2-(N-吗啉代)乙磺酸）。使用两个参考比使用一个参考可以更好地定义峰体积和浓度之间的线性关系，并且测量的混合物中单个化合物的绝对浓度更准确。我们将半自动 gsHSQC0 的结果与原始手动阶段循环 HSQC0 方法获得的结果进行比较。新方法适用于通过同时定量复杂混合物中的多种代谢物来自动进行代谢物分析。
Time-zero 2D 13C HSQC (HSQC0) spectroscopy offers advantages over traditional 2D NMR for quantitative analysis of solutions containing a mixture of compounds because the signal intensities are directly proportional to the concentrations of the constituents. The HSQC0 spectrum is derived from a series of spectra collected with increasing repetition times within the basic HSQC block by extrapolating the repetition time to zero. Here we present an alternative approach to data collection, gradient-selective time-zero 1H-13C HSQC0 in combination with fast maximum likelihood reconstruction (FMLR) data analysis and the use of two concentration references for absolute concentration determination. Gradient-selective data acquisition results in cleaner spectra, and NMR data can be acquired in both constant-time and non-constant time mode. Semi-automatic data analysis is supported by the FMLR approach, which is used to deconvolute the spectra and extract peak volumes. The peak volumes obtained from this analysis are converted to absolute concentrations by reference to the peak volumes of two internal reference compounds of known concentration: DSS (4,4-dimethyl-4-silapentane-1-sulfonic acid) at the low concentration limit (which also serves as chemical shift reference) and MES (2-(N-morpholino)ethanesulfonic acid) at the high concentration limit. The linear relationship between peak volumes and concentration is better defined with two references than with one, and the measured absolute concentrations of individual compounds in the mixture are more accurate. We compare results from semi-automated gsHSQC0 with those obtained by the original manual phase-cycled HSQC0 approach. The new approach is suitable for automatic metabolite profiling by simultaneous quantification of multiple metabolites in a complex mixture.
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通过二维外推时间零（1）H-（13）C HSQC（HSQC（0））同时定量和鉴定代谢物混合物中的单个化学物质。
影响因子：
15
作者：
Hu K;Westler WM;Markley JL
通讯作者：
Markley JL
影响因子：
7.4
作者：
Akoka, S;Barantin, L;Trierweiler, M
通讯作者：
Trierweiler, M
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：
2.7
作者：
Chylla, RA;Volkman, BF;Markley, JL
通讯作者：
Markley, JL
影响因子：
5.1
作者：
Hu K;Wyche TP;Bugni TS;Markley JL
通讯作者：
Markley JL