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

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

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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. Abstract Text:

Hui Wei,Kerstin Nolkrantz,Mark C Parkin,Claire N Chisolm,James P O'Callaghan,Robert T Kennedy,

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 15N3-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.

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

H Wei,K Nolkrantz,MC Parkin,CN Chisolm,JP O'Callaghan,RT Kennedy,

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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. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Analytical chemistry

VOLUME: 78

Page Numbers: 4342-51

Journal Abbreviation: Anal. Chem.

ISSN: 0003-2700

DAY: 1

MONTH: Jul

YEAR: 2006

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

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LANGUAGE: eng

NlmUniqueID: 370536

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

KEYWORDS: Spectrometry, Mass, Matrix-Assisted Lase

MESH TERMS: methods

Chemical & Substance for Abstract: Identification and quantification of neuropeptides in brain tissue by capillary liquid chromatography coupled off-line to MALDI-TOF and MALDI-TOF/TOF-MS. Information

Substance Name: Neuropeptides

Registry Number: 0

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

AFFILIATION: Department of Chemistry and Department of Pharmacology, University of Michigan, Ann Arbor, Michigan 48109, USA.

Country: United States

AGENCY: United States NINDS

GRANT: NS 38476

ACRONYM: NS

MEDLINETA: Anal Chem

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