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 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.
Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy.
Andrea
A
Armirotti
Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy.
Abdul
A
Basit
Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy.
Natalia
N
Realini
Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia, Via Ardeatina 306, 00179 Rome, Italy.
Carlo
C
Caltagirone
Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia, Via Ardeatina 306, 00179 Rome, Italy.
Paola
P
Bossù
Department of Clinical and Behavioral Neurology, IRCCS Fondazione Santa Lucia, Via Ardeatina 306, 00179 Rome, Italy.
Gianfranco
G
Spalletta
Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy; Department of Anatomy and Neurobiology, University of California, Irvine, CA 92697, USA; Department of Pharmacology, University of California, Irvine, CA 92697, USA; Department of Biological Chemistry, University of California, Irvine, CA 92697, USA. Electronic address: piomelli@uci.edu.
Daniele
D
Piomelli
D000368:Aged; D000544:Alzheimer Disease / Q000097:blood; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D001798:Blood Proteins / Q000032:analysis*; D016022:Case-Control Studies; D002853:Chromatography, Liquid / Q000379:methods*; D003704:Dementia / Q000097:blood*; D005260:Female; D006801:Humans; D008297:Male; D055432:Metabolomics / Q000379:methods; D051379:Mice; D015999:Multivariate Analysis; D013109:Sphingomyelins / Q000097:blood; D053719:Tandem Mass Spectrometry
D001798:Blood Proteins; D013109:Sphingomyelins
Dried plasma spot; Mass spectrometry; Metabolomics; Orthogonal chromatography; Sample preparation