Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood
Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1<span style="font-variant: small-caps;">d</span>-<sup>1</sup>H NMR spectra and disrupt quantification of metabolites. Here, we present effect...
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doaj-b995de571dbb43f7848601a6384513102020-11-24T21:22:13ZengMDPI AGMetabolites2218-19892018-12-01849310.3390/metabo8040093metabo8040093Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole BloodCora E. McHugh0Thomas L. Flott1Casey R. Schooff2Zyad Smiley3Michael A. Puskarich4Daniel D. Myers5John G. Younger6Alan E. Jones7Kathleen A. Stringer8NMR Metabolomics Laboratory, Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USANMR Metabolomics Laboratory, Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USANMR Metabolomics Laboratory, Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USANMR Metabolomics Laboratory, Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USADepartment of Emergency Medicine, University of Minnesota, Minneapolis, MN 55455, USAUnit for Laboratory Animal Medicine, University of Michigan, Ann Arbor, MI 48109, USAAkadeum Life Sciences, Ann Arbor, MI 48103, USADepartment of Emergency Medicine, University of Mississippi, Jackson, MS 39216, USANMR Metabolomics Laboratory, Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USABackground: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1<span style="font-variant: small-caps;">d</span>-<sup>1</sup>H NMR spectra and disrupt quantification of metabolites. Here, we present effective macromolecule removal strategies for serum and whole blood (WB) samples. Methods: A variety of macromolecule removal strategies were compared in both WB and serum, along with tests of ultrafiltration alone and in combination with precipitation methods. Results: In healthy human serum, methanol:chloroform:water extraction with ultrafiltration was compared to methanol precipitation with and without ultrafiltration. Methods were tested in healthy pooled human serum, and in serum from patients with sepsis. Effects of long-term storage at −80 °C were tested to explore the impact of macromolecule removal strategy on serum from different conditions. In WB a variety of extraction strategies were tested in two types of WB (from pigs and baboons) to examine the impact of macromolecule removal strategies on different samples. Conclusions: In healthy human serum methanol precipitation of serum with ultrafiltration was superior, but was similar in recovery and variance to methanol:chloroform:water extraction with ultrafiltration in pooled serum from patients with sepsis. In WB, high quality, quantifiable spectra were obtained with the use of a methanol: chloroform precipitation.https://www.mdpi.com/2218-1989/8/4/93pharmacometabolomicsextractionultrafiltration1<span style="font-variant: small-caps">d</span>-<sup>1</sup>H NMRquantitative analysispreanalytical processing |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Cora E. McHugh Thomas L. Flott Casey R. Schooff Zyad Smiley Michael A. Puskarich Daniel D. Myers John G. Younger Alan E. Jones Kathleen A. Stringer |
spellingShingle |
Cora E. McHugh Thomas L. Flott Casey R. Schooff Zyad Smiley Michael A. Puskarich Daniel D. Myers John G. Younger Alan E. Jones Kathleen A. Stringer Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood Metabolites pharmacometabolomics extraction ultrafiltration 1<span style="font-variant: small-caps">d</span>-<sup>1</sup>H NMR quantitative analysis preanalytical processing |
author_facet |
Cora E. McHugh Thomas L. Flott Casey R. Schooff Zyad Smiley Michael A. Puskarich Daniel D. Myers John G. Younger Alan E. Jones Kathleen A. Stringer |
author_sort |
Cora E. McHugh |
title |
Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood |
title_short |
Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood |
title_full |
Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood |
title_fullStr |
Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood |
title_full_unstemmed |
Rapid, Reproducible, Quantifiable NMR Metabolomics: Methanol and Methanol: Chloroform Precipitation for Removal of Macromolecules in Serum and Whole Blood |
title_sort |
rapid, reproducible, quantifiable nmr metabolomics: methanol and methanol: chloroform precipitation for removal of macromolecules in serum and whole blood |
publisher |
MDPI AG |
series |
Metabolites |
issn |
2218-1989 |
publishDate |
2018-12-01 |
description |
Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1<span style="font-variant: small-caps;">d</span>-<sup>1</sup>H NMR spectra and disrupt quantification of metabolites. Here, we present effective macromolecule removal strategies for serum and whole blood (WB) samples. Methods: A variety of macromolecule removal strategies were compared in both WB and serum, along with tests of ultrafiltration alone and in combination with precipitation methods. Results: In healthy human serum, methanol:chloroform:water extraction with ultrafiltration was compared to methanol precipitation with and without ultrafiltration. Methods were tested in healthy pooled human serum, and in serum from patients with sepsis. Effects of long-term storage at −80 °C were tested to explore the impact of macromolecule removal strategy on serum from different conditions. In WB a variety of extraction strategies were tested in two types of WB (from pigs and baboons) to examine the impact of macromolecule removal strategies on different samples. Conclusions: In healthy human serum methanol precipitation of serum with ultrafiltration was superior, but was similar in recovery and variance to methanol:chloroform:water extraction with ultrafiltration in pooled serum from patients with sepsis. In WB, high quality, quantifiable spectra were obtained with the use of a methanol: chloroform precipitation. |
topic |
pharmacometabolomics extraction ultrafiltration 1<span style="font-variant: small-caps">d</span>-<sup>1</sup>H NMR quantitative analysis preanalytical processing |
url |
https://www.mdpi.com/2218-1989/8/4/93 |
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