Wallmeier, Jens and Samol, Claudia and Ellmann, Lisa and Zacharias, Helena U. and Vogl, Franziska C. and Garcia, Muriel and Dettmer, Katja and Oefner, Peter J. and Gronwald, Wolfram (2017) Quantification of Metabolites by NMR Spectroscopy in the Presence of Protein. JOURNAL OF PROTEOME RESEARCH, 16 (4). pp. 1784-1796. ISSN 1535-3893, 1535-3907
Full text not available from this repository. (Request a copy)Abstract
The high reliability of NMR spectroscopy makes it an ideal tool for large-scale metabolomic studies. However, the complexity of biofluids and, in particular, the presence of macromolecules poses a significant challenge. Ultrafiltration and protein precipitation are established means of deproteinization and recovery of free or total metabolite content, but neither is ever complete. In addition, aside from cost and labor, all deproteinization methods constitute an additional source of experimental variation. The Carr-Purcell-Meiboom-Gill (CPMG) echo-train acquisition of NMR spectra obviates the need for prior deproteinization by attenuating signals from macromolecules, but concentration values of metabolites measured in blood plasma will not necessarily reflect total or free metabolite content. Here, in contrast to approaches that propose the determination of individual T1 and T2 relaxation times for the computation of correction factors, we demonstrate their determination by spike-in experiments with known amounts of metabolites in pooled samples of the matrix of interest to facilitate the measurement of total metabolite content. Provided that the protein content does not vary too much among individual samples, accurate quantitation of metabolites is feasible. Moreover, samples with significantly deviating protein content may be readily recognized by inclusion of a standard that shows moderate protein binding. It is also shown that urinary proteins when present in high concentrations may effect detection of common urinary metabolites prone to strong protein binding such as tryptophan.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MAGNETIC-RESONANCE-SPECTROSCOPY; H-1-NMR SPECTROSCOPY; NEPHROTIC SYNDROME; BLOOD-PLASMA; SERUM; H-1; METABOLOMICS; METABONOMICS; TRYPTOPHAN; EXTRACTS; ID NMR; quantification; plasma; CPMG; correction factors |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Institut für Funktionelle Genomik > Lehrstuhl für Funktionelle Genomik (Prof. Oefner) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 13:11 |
| Last Modified: | 20 Feb 2019 07:31 |
| URI: | https://pred.uni-regensburg.de/id/eprint/1122 |
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