Rapid, reproducible, quantifiable nmr metabolomics: Methanol and methanol: Chloroform precipitation for removal of macromolecules in serum and whole blood

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

doi:10.3390/metabo8040093

Rapid, reproducible, quantifiable nmr metabolomics: Methanol and methanol: Chloroform precipitation for removal of macromolecules in serum and whole blood

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

Emergency Medicine

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1D-1H 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.

Original language	English (US)
Article number	93
Journal	Metabolites
Volume	8
Issue number	4
DOIs	https://doi.org/10.3390/metabo8040093
State	Published - Dec 2018

Bibliographical note

Funding Information:
Funding: This research was funded by grants from the National Institute of General Medical Sciences (NIGMS), National Institutes of Health (NIH) to KAS (R01 GM111400) and metabolomics supplements to (R01 GM069438, JGY) and (R01 GM103799, AEJ). The content is solely the responsibility of the authors and does not necessarily present the official views of the NIGMS or the National Institutes of Health.

Publisher Copyright:
© 2018 by the authors.

Keywords

1d-1h nmr
Extraction
Pharmacometabolomics
Preanalytical processing
Quantitative analysis
Ultrafiltration

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@article{b64405cc4adb4bcdaea75426b297edca,

title = "Rapid, reproducible, quantifiable nmr metabolomics: Methanol and methanol: Chloroform precipitation for removal of macromolecules in serum and whole blood",

abstract = "Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1D-1H 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.",

keywords = "1d-1h nmr, Extraction, Pharmacometabolomics, Preanalytical processing, Quantitative analysis, Ultrafiltration",

author = "McHugh, {Cora E.} and Flott, {Thomas L.} and Schooff, {Casey R.} and Zyad Smiley and Puskarich, {Michael A.} and Myers, {Daniel D.} and Younger, {John G.} and Jones, {Alan E.} and Stringer, {Kathleen A.}",

note = "Funding Information: Funding: This research was funded by grants from the National Institute of General Medical Sciences (NIGMS), National Institutes of Health (NIH) to KAS (R01 GM111400) and metabolomics supplements to (R01 GM069438, JGY) and (R01 GM103799, AEJ). The content is solely the responsibility of the authors and does not necessarily present the official views of the NIGMS or the National Institutes of Health. Publisher Copyright: {\textcopyright} 2018 by the authors.",

year = "2018",

month = dec,

doi = "10.3390/metabo8040093",

language = "English (US)",

volume = "8",

journal = "Metabolites",

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T1 - Rapid, reproducible, quantifiable nmr metabolomics

T2 - Methanol and methanol: Chloroform precipitation for removal of macromolecules in serum and whole blood

AU - McHugh, Cora E.

AU - Flott, Thomas L.

AU - Schooff, Casey R.

AU - Smiley, Zyad

AU - Puskarich, Michael A.

AU - Myers, Daniel D.

AU - Younger, John G.

AU - Jones, Alan E.

AU - Stringer, Kathleen A.

N1 - Funding Information: Funding: This research was funded by grants from the National Institute of General Medical Sciences (NIGMS), National Institutes of Health (NIH) to KAS (R01 GM111400) and metabolomics supplements to (R01 GM069438, JGY) and (R01 GM103799, AEJ). The content is solely the responsibility of the authors and does not necessarily present the official views of the NIGMS or the National Institutes of Health. Publisher Copyright: © 2018 by the authors.

PY - 2018/12

Y1 - 2018/12

N2 - Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1D-1H 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.

AB - Background: Though blood is an excellent biofluid for metabolomics, proteins and lipids present in blood can interfere with 1D-1H 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.

KW - 1d-1h nmr

KW - Extraction

KW - Pharmacometabolomics

KW - Preanalytical processing

KW - Quantitative analysis

KW - Ultrafiltration

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U2 - 10.3390/metabo8040093

DO - 10.3390/metabo8040093

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C2 - 30558115

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SN - 2218-1989

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JO - Metabolites

JF - Metabolites

IS - 4

M1 - 93

ER -

Rapid, reproducible, quantifiable nmr metabolomics: Methanol and methanol: Chloroform precipitation for removal of macromolecules in serum and whole blood

Abstract

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Keywords

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