Cargando…

Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments

The spectral resolution of 2D [Formula: see text] H- [Formula: see text] C heteronuclear single quantum coherence ([Formula: see text] H- [Formula: see text] C-HSQC) nuclear magnetic resonance (NMR) spectra facilitates both metabolite identification and quantification in nuclear magnetic resonance-b...

Descripción completa

Detalles Bibliográficos
Autores principales: Weitzel, Alexander, Samol, Claudia, Oefner, Peter J., Gronwald, Wolfram
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7695005/
https://www.ncbi.nlm.nih.gov/pubmed/33171777
http://dx.doi.org/10.3390/metabo10110449
_version_ 1783615100928655360
author Weitzel, Alexander
Samol, Claudia
Oefner, Peter J.
Gronwald, Wolfram
author_facet Weitzel, Alexander
Samol, Claudia
Oefner, Peter J.
Gronwald, Wolfram
author_sort Weitzel, Alexander
collection PubMed
description The spectral resolution of 2D [Formula: see text] H- [Formula: see text] C heteronuclear single quantum coherence ([Formula: see text] H- [Formula: see text] C-HSQC) nuclear magnetic resonance (NMR) spectra facilitates both metabolite identification and quantification in nuclear magnetic resonance-based metabolomics. However, quantification is complicated by variations in magnetization transfer, which among others originate mainly from scalar coupling differences. Methods that compensate for variation in scalar coupling include the generation of calibration factors for individual signals or the use of additional pulse sequence schemes such as quantitative HSQC (Q-HSQC) that suppress the J(CH)-dependence by modulating the polarization transfer delays of HSQC or, additionally, employ a pure-shift homodecoupling approach in the [Formula: see text] H dimension, such as Quantitative, Perfected and Pure Shifted HSQC (QUIPU-HSQC). To test the quantitative accuracy of these three methods, employing a 600 MHz NMR spectrometer equipped with a helium cooled cryoprobe, a Latin-square design that covered the physiological concentration ranges of 10 metabolites was used. The results show the suitability of all three methods for the quantification of highly abundant metabolites. However, the substantially increased residual water signal observed in QUIPU-HSQC spectra impeded the quantification of low abundant metabolites located near the residual water signal, thus limiting its utility in high-throughput metabolite fingerprinting studies.
format Online
Article
Text
id pubmed-7695005
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-76950052020-11-28 Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments Weitzel, Alexander Samol, Claudia Oefner, Peter J. Gronwald, Wolfram Metabolites Article The spectral resolution of 2D [Formula: see text] H- [Formula: see text] C heteronuclear single quantum coherence ([Formula: see text] H- [Formula: see text] C-HSQC) nuclear magnetic resonance (NMR) spectra facilitates both metabolite identification and quantification in nuclear magnetic resonance-based metabolomics. However, quantification is complicated by variations in magnetization transfer, which among others originate mainly from scalar coupling differences. Methods that compensate for variation in scalar coupling include the generation of calibration factors for individual signals or the use of additional pulse sequence schemes such as quantitative HSQC (Q-HSQC) that suppress the J(CH)-dependence by modulating the polarization transfer delays of HSQC or, additionally, employ a pure-shift homodecoupling approach in the [Formula: see text] H dimension, such as Quantitative, Perfected and Pure Shifted HSQC (QUIPU-HSQC). To test the quantitative accuracy of these three methods, employing a 600 MHz NMR spectrometer equipped with a helium cooled cryoprobe, a Latin-square design that covered the physiological concentration ranges of 10 metabolites was used. The results show the suitability of all three methods for the quantification of highly abundant metabolites. However, the substantially increased residual water signal observed in QUIPU-HSQC spectra impeded the quantification of low abundant metabolites located near the residual water signal, thus limiting its utility in high-throughput metabolite fingerprinting studies. MDPI 2020-11-07 /pmc/articles/PMC7695005/ /pubmed/33171777 http://dx.doi.org/10.3390/metabo10110449 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Weitzel, Alexander
Samol, Claudia
Oefner, Peter J.
Gronwald, Wolfram
Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments
title Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments
title_full Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments
title_fullStr Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments
title_full_unstemmed Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments
title_short Robust Metabolite Quantification from J-Compensated 2D (1)H-(13)C-HSQC Experiments
title_sort robust metabolite quantification from j-compensated 2d (1)h-(13)c-hsqc experiments
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7695005/
https://www.ncbi.nlm.nih.gov/pubmed/33171777
http://dx.doi.org/10.3390/metabo10110449
work_keys_str_mv AT weitzelalexander robustmetabolitequantificationfromjcompensated2d1h13chsqcexperiments
AT samolclaudia robustmetabolitequantificationfromjcompensated2d1h13chsqcexperiments
AT oefnerpeterj robustmetabolitequantificationfromjcompensated2d1h13chsqcexperiments
AT gronwaldwolfram robustmetabolitequantificationfromjcompensated2d1h13chsqcexperiments