Journal
JOURNAL OF MASS SPECTROMETRY
Volume 49, Issue 8, Pages 665-673Publisher
WILEY
DOI: 10.1002/jms.3411
Keywords
MALDI imaging; in-tissue derivatization; amine metabolites; amino acids; neurotransmitters; identification; reagent precoated MALDI targets
Funding
- National Institutes of Health, NIH/NIGMS [5GM 058008-15, 5P41 103391-04]
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Imaging mass spectrometry (IMS) studies increasingly focus on endogenous small molecular weight metabolites and consequently bring special analytical challenges. Since analytical tissue blanks do not exist for endogenous metabolites, careful consideration must be given to confirm molecular identity. Here, we present approaches for the improvement in detection of endogenous amine metabolites such as amino acids and neurotransmitters in tissues through chemical derivatization and matrix-assisted laser desorption/ionization (MALDI) IMS. Chemical derivatization with 4-hydroxy-3-methoxycinnamaldehyde (CA) was used to improve sensitivity and specificity. CA was applied to the tissue via MALDI sample targets precoated with a mixture of derivatization reagent and ferulic acid as a MALDI matrix. Spatial distributions of chemically derivatized endogenous metabolites in tissue were determined by high-mass resolution and MSn IMS. We highlight an analytical strategy for metabolite validation whereby tissue extracts are analyzed by high-performance liquid chromatography (HPLC)-MS/MS to unambiguously identify metabolites and distinguish them from isobaric compounds. Copyright (C) 2014 John Wiley & Sons, Ltd.
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