Sample preparation and orthogonal chromatography for broad polarity range plasma metabolomics: Application to human subjects with neurodegenerative dementia

We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC–MS/MS analysis of mouse and human plasma samples, which enables the untargeted (“shotgun”) or targeted profiling of hydrophilic, amphipathic, and hydrophobic constituents of plasma metabolome. The protocol is...

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Published in	Analytical biochemistry Vol. 455; pp. 48 - 54
Main Authors	Armirotti, Andrea, Basit, Abdul, Realini, Natalia, Caltagirone, Carlo, Bossù, Paola, Spalletta, Gianfranco, Piomelli, Daniele
Format	Journal Article
Language	English
Published	United States Elsevier Inc 15.06.2014
Subjects	Aged Alzheimer disease Alzheimer Disease - blood Animals Biochemistry - methods Blood Proteins - analysis Case-Control Studies chromatography Chromatography, Liquid - methods cognitive disorders Dementia - blood Dried plasma spot Female Humans hydrophilicity hydrophobicity Male Mass spectrometry metabolome Metabolomics Metabolomics - methods Mice Multivariate Analysis Orthogonal chromatography Sample preparation Sphingomyelins - blood Tandem Mass Spectrometry Metabolomics Dried plasma spot Sample preparation Mass spectrometry Orthogonal chromatography
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ISSN	0003-2697 1096-0309 1096-0309
DOI	10.1016/j.ab.2014.03.019

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Abstract	We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC–MS/MS analysis of mouse and human plasma samples, which enables the untargeted (“shotgun”) or targeted profiling of hydrophilic, amphipathic, and hydrophobic constituents of plasma metabolome. The protocol is rapid, efficient, and reliable, and offers several advantages compared to current procedures. When applied to a training set of human plasma samples, the protocol allowed for the rapid acquisition of full LogP metabolic profiles in plasma samples obtained from cognitively healthy human subjects and age-matched subjects with mild cognitive impairment or Alzheimer’s disease (n=15 each). Targeted analyses confirmed these findings, which are consistent with data previously published by other groups.
AbstractList	We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC-MS/MS analysis of mouse and human plasma samples, which enables the untargeted ("shotgun") or targeted profiling of hydrophilic, amphipathic, and hydrophobic constituents of plasma metabolome. The protocol is rapid, efficient, and reliable, and offers several advantages compared to current procedures. When applied to a training set of human plasma samples, the protocol allowed for the rapid acquisition of full LogP metabolic profiles in plasma samples obtained from cognitively healthy human subjects and age-matched subjects with mild cognitive impairment or Alzheimer's disease (n=15 each). Targeted analyses confirmed these findings, which are consistent with data previously published by other groups.We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC-MS/MS analysis of mouse and human plasma samples, which enables the untargeted ("shotgun") or targeted profiling of hydrophilic, amphipathic, and hydrophobic constituents of plasma metabolome. The protocol is rapid, efficient, and reliable, and offers several advantages compared to current procedures. When applied to a training set of human plasma samples, the protocol allowed for the rapid acquisition of full LogP metabolic profiles in plasma samples obtained from cognitively healthy human subjects and age-matched subjects with mild cognitive impairment or Alzheimer's disease (n=15 each). Targeted analyses confirmed these findings, which are consistent with data previously published by other groups. We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC-MS/MS analysis of mouse and human plasma samples, which enables the untargeted ("shotgun") or targeted profiling of hydrophilic, amphipathic, and hydrophobic constituents of plasma metabolome. The protocol is rapid, efficient, and reliable, and offers several advantages compared to current procedures. When applied to a training set of human plasma samples, the protocol allowed for the rapid acquisition of full LogP metabolic profiles in plasma samples obtained from cognitively healthy human subjects and age-matched subjects with mild cognitive impairment or Alzheimer's disease (n=15 each). Targeted analyses confirmed these findings, which are consistent with data previously published by other groups.
Author	Piomelli, Daniele Realini, Natalia Caltagirone, Carlo Bossù, Paola Armirotti, Andrea Basit, Abdul Spalletta, Gianfranco
Author_xml	– sequence: 1 givenname: Andrea surname: Armirotti fullname: Armirotti, Andrea organization: Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy – sequence: 2 givenname: Abdul surname: Basit fullname: Basit, Abdul organization: Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy – sequence: 3 givenname: Natalia surname: Realini fullname: Realini, Natalia organization: Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy – sequence: 4 givenname: Carlo surname: Caltagirone fullname: Caltagirone, Carlo organization: Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia, Via Ardeatina 306, 00179 Rome, Italy – sequence: 5 givenname: Paola surname: Bossù fullname: Bossù, Paola organization: Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia, Via Ardeatina 306, 00179 Rome, Italy – sequence: 6 givenname: Gianfranco surname: Spalletta fullname: Spalletta, Gianfranco organization: Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia, Via Ardeatina 306, 00179 Rome, Italy – sequence: 7 givenname: Daniele surname: Piomelli fullname: Piomelli, Daniele email: piomelli@uci.edu organization: Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy
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Snippet	We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC–MS/MS analysis of mouse and human plasma samples, which enables the... We describe a simple protocol for the preparation and orthogonal hydrophobic/hydrophilic LC-MS/MS analysis of mouse and human plasma samples, which enables the...
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SubjectTerms	Aged Alzheimer disease Alzheimer Disease - blood Animals Biochemistry - methods Blood Proteins - analysis Case-Control Studies chromatography Chromatography, Liquid - methods cognitive disorders Dementia - blood Dried plasma spot Female Humans hydrophilicity hydrophobicity Male Mass spectrometry metabolome Metabolomics Metabolomics - methods Mice Multivariate Analysis Orthogonal chromatography Sample preparation Sphingomyelins - blood Tandem Mass Spectrometry
Title	Sample preparation and orthogonal chromatography for broad polarity range plasma metabolomics: Application to human subjects with neurodegenerative dementia
URI	https://dx.doi.org/10.1016/j.ab.2014.03.019 https://www.ncbi.nlm.nih.gov/pubmed/24708938 https://www.proquest.com/docview/1523404039 https://www.proquest.com/docview/2101378376
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