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Identification and quantification of neuropeptides in brain tissue by capillary liquid chromatography coupled off-line to MALDI-TOF and MALDI-TOF/TOF-MS.

Authors
Wei-H; Nolkrantz-K; Parkin-MC; Chisolm-CN; O'Callaghan-JP; Kennedy-RT
Source
Anal Chem 2006 Jul; 78(13):4342-4351
NIOSHTIC No.
20030622
Abstract
Capillary liquid chromatography (CLC) coupled off-line with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and TOF/TOF-MS were explored for identification and quantification of neuropeptides in microwave-fixed rat brain tissue. Sample was separated by gradient elution on 50-mum-inner diameter reversed-phase columns at 180 nL/min. Effluent was mixed with matrix solution and transferred to a MALDI target plate by pulsed electric field deposition, yielding sample spots with 200-300-mum diameter. Mass detection limits as low as 2 amol, corresponding to 1 pM concentration, were achieved for neuropeptides. CLC-MALDI-TOF-MS analysis of microwave-fixed rat striatum tissue yielded detection of over 400 distinctive peaks. CLC-MALDI-TOF/TOF-MS allowed identification of 10 peptides including 3 novel peptides. Quantification was evaluated using substance P as analyte and (15)N(3)-labeled substance P as an internal standard. Quantification of substance P revealed approximately 6.8-fold higher levels than previously reported in the rat striatum. This increase is attributed to use of microwave fixation, which prevented degradation of the peptide, aggressive extraction procedures, and accounting for oxidation of substance P in the analysis. These results demonstrate that CLC-MALDI-TOF-MS is a versatile tool for neuropeptide analysis in brain tissue by allowing for detection, identification, and quantification.
Keywords
Brain-matter; Ionization; Mass-spectrometry; Laboratory-animals; Animals; Animal-studies; Sampling; Sampling-methods; Peptides; Liquid-chromatography; Sample-preparation; Lasers
CODEN
ANCHAM
Publication Date
20060701
Document Type
Journal Article
Fiscal Year
2006
NTIS Accession No.
NTIS Price
Issue of Publication
13
ISSN
0003-2700
NIOSH Division
HELD
Source Name
Analytical Chemistry
State
MI
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