Quantitative 1H NMR spectroscopy

Bharti, Santosh Kumar; Roy, Raja

doi:10.1016/j.trac.2012.02.007

Cited by 617 publications

(595 citation statements)

References 46 publications

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“…The concentration of analytes obtained after lyophilization was identical to theoretical values, within the inaccuracy of the PULCON method. By contrast, as expected [4], inaccuracy was higher, especially for the b-glucose signal, when solvent suppression was used (see Table S2 in supplementary material).…”

Section: Methods Validationsupporting

confidence: 53%

“…Methanol, acetonitrile, deuterium oxide (D 2 O), monobasic (KH 2 PO 4 ) and dibasic (K 2 HPO 4 ) potassium phosphate, dimethylsulfone (DMS), Dulbecco's modified Eagle's medium (DMEM), and 3-(trimethylsilyl)propionic-2,2,3,3-d 4 acid sodium salt (TSP) were obtained from SigmaeAldrich (Milan, Italy).…”

Section: Methodsmentioning

confidence: 99%

“…An exponential line broadening (0.1 Hz) was applied to FIDs (free induction decays) before Fourier transform. The spectra were manually phased and automatically baseline corrected (as preferred in metabolomics [4] …”

Section: Nmr Spectroscopymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…Compared with other analytical techniques, q-NMR does not require chromatographic separation, generates signals that are directly proportional to the number of NMR-active nuclei in the targeted analyte [4,5], and offers a high degree of assay reproducibility [6] along with reduced uncertainty [7,8]. Moreover, advances in hardware developmentdsuch as the introduction of high-field magnets and cryogenic probesdhave progressively lowered the limit of detection (LOD) for analytes, thereby improving the overall sensitivity of the technique [4].The PULCON (pulse length-based concentration determination) procedure [9], one of the most promising methods for q-NMR [4], finds its mathematical and physical foundation in the principle of reciprocity [10e12]. This principle states that the strength of a signal is inversely proportional to the 90 pulse length in the active volume of the NMR tube.…”

mentioning

confidence: 99%

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A simple and accurate protocol for absolute polar metabolite quantification in cell cultures using quantitative nuclear magnetic resonance

Goldoni

Beringhelli

Rocchia

et al. 2016

Analytical Biochemistry

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a b s t r a c tAbsolute analyte quantification by nuclear magnetic resonance (NMR) spectroscopy is rarely pursued in metabolomics, even though this would allow researchers to compare results obtained using different techniques. Here we report on a new protocol that permits, after pH-controlled serum protein removal, the sensitive quantification (limit of detection [LOD] ¼ 5À25 mM) of hydrophilic nutrients and metabolites in the extracellular medium of cells in cultures. The method does not require the use of databases and uses PULCON (pulse length-based concentration determination) quantitative NMR to obtain results that are significantly more accurate and reproducible than those obtained by CPMG (CarrePurcell eMeiboomeGill) sequence or post-processing filtering approaches. Three practical applications of the method highlight its flexibility under different cell culture conditions. We identified and quantified (i) metabolic differences between genetically engineered human cell lines, (ii) alterations in cellular metabolism induced by differentiation of mouse myoblasts into myotubes, and (iii) metabolic changes caused by activation of neurotransmitter receptors in mouse myoblasts. Thus, the new protocol offers an easily implementable, efficient, and versatile tool for the investigation of cellular metabolism and signal transduction.© 2016 Elsevier Inc. All rights reserved.Interest in quantitative nuclear magnetic resonance (q-NMR) analysis has steadily increased since the seminal works of Jungnickel and Forbes [1] and Hollis [2], owing to the unique ability of this methodological approach to quantify, at the same time, a wide range of structurally diverse biological substances with high throughput and automation [3]. Compared with other analytical techniques, q-NMR does not require chromatographic separation, generates signals that are directly proportional to the number of NMR-active nuclei in the targeted analyte [4,5], and offers a high degree of assay reproducibility [6] along with reduced uncertainty [7,8]. Moreover, advances in hardware developmentdsuch as the introduction of high-field magnets and cryogenic probesdhave progressively lowered the limit of detection (LOD) for analytes, thereby improving the overall sensitivity of the technique [4].The PULCON (pulse length-based concentration determination) procedure [9], one of the most promising methods for q-NMR [4], finds its mathematical and physical foundation in the principle of reciprocity [10e12]. This principle states that the strength of a signal is inversely proportional to the 90 pulse length in the active volume of the NMR tube. The PULCON method can be implemented in all types of commercial NMR spectrometers and requires only one Abbreviations used: q-NMR, quantitative nuclear magnetic resonance; LOD, limit of detection; PULCON, pulse length-based concentration determination; CPMG,

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Section: Methods Validationsupporting

confidence: 53%

Section: Methodsmentioning

confidence: 99%

Section: Nmr Spectroscopymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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A simple and accurate protocol for absolute polar metabolite quantification in cell cultures using quantitative nuclear magnetic resonance

Goldoni

Beringhelli

Rocchia

et al. 2016

Analytical Biochemistry

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Metabolomics: Experimental Design, Methodology and Data Analysis

Schripsema

Dagnino

2014

Encyclopedia of Analytical Chemistry

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Metabolomics can be defined as the area of research that strives to obtain complete metabolic fingerprints, to detect differences between them and to provide hypothesis to explain those differences. This area of research is still rapidly developing into a powerful tool in the study of all types of organisms. This chapter focuses on the practical and efficient application of metabolomics. All different aspects of a metabolomic study are discussed, ranging from the initial questions and experimental design to the instrumental methods and data analysis.

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Metabolomics: Experimental Design, Methodology, and Data Analysis

Schripsema¹,

Dagnino²

2019

Encyclopedia of Analytical Chemistry

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This article focuses on the practical and potential applications of metabolomics. The different steps of metabolomics experiments are discussed, from the initial design, sampling, and measurements, to chromatograms and spectra, pretreatment of data, and interpretation of data to find marker compounds. The article discusses how to choose the analytical techniques, extraction procedures, and methods for data analysis. The techniques in metabolomics are increasingly standardized, but rapid development is still going on, leading to the introduction of new techniques and methods. The latest developments have been reviewed.

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Quantitative 1H NMR spectroscopy

Cited by 617 publications

References 46 publications

A simple and accurate protocol for absolute polar metabolite quantification in cell cultures using quantitative nuclear magnetic resonance

A simple and accurate protocol for absolute polar metabolite quantification in cell cultures using quantitative nuclear magnetic resonance

Metabolomics: Experimental Design, Methodology and Data Analysis

Metabolomics: Experimental Design, Methodology, and Data Analysis

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