Analysis of substance P in rat brain by means of immunoaffinity capture and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry

Interest in the analysis of low abundance neuropeptides particularly using matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI–TOF–MS) is increasing because these neuropeptides are essential to the mechanism of transportation and the metabolism. This article describes...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2004-08, Vol.807 (2), p.307-313
Main Authors: Suresh Babu, C.V, Lee, Jeongae, Lho, Dong Seok, Yoo, Young Sook
Format: Article
Language:English
Subjects:
Analysis
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Brain - metabolism
Calibration
Electrophoresis, Capillary
Fundamental and applied biological sciences. Psychology
General pharmacology
Immunoprecipitation
Medical sciences
Pharmacology. Drug treatments
Rats
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
Substance P
Substance P - metabolism
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Summary:Interest in the analysis of low abundance neuropeptides particularly using matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI–TOF–MS) is increasing because these neuropeptides are essential to the mechanism of transportation and the metabolism. This article describes an immunoprecipitation procedure that is suitable for MALDI–MS analysis of substance P (SP), a neuropeptide, in rat brain tissues. Substance P was precipitated from brain tissue extracts by immunoprecipitation with antibodies directed against SP, and are analyzed by MALDI–TOF–MS. Mass spectrometric analysis showed a singly charged [ M+H] + ion peak that corresponded to the SP molecular mass and was observed with a detection error of 1.6%. The average mass errors between the observed and theoretical molecular mass were within the 0.11 Da range. Capillary zone electrophoresis analysis was subsequently performed, and the effects of the different separation parameters were examined. Beginning with milligram quantities of brain tissue, picomole quantities of SP could be detected using this method.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2004.04.026