Journal
JOURNAL OF MASS SPECTROMETRY
Volume 40, Issue 10, Pages 1338-1346Publisher
WILEY
DOI: 10.1002/jms.916
Keywords
matrix-assisted laser desorption/ionization mass spectrometry; neuropeptides; microwave-fixed brain tissue; octadecanethiol self-assembled monolayer; nanocolumn
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A sample preparation method that combines a modified target plate with a nanoscale reversed-phase column (nanocolumn) was developed for detection of neuropeptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A gold-coated MALDI plate was modified with an octadecanethiol (ODT) self-assembled monolayer to create a hydrophobic surface that could concentrate peptide samples into a similar to 200-500-mu m diameter spot. The spot sizes generated were comparable to those obtained for a substrate patterned with 200-mu m hydrophilic spots on a hydrophobic substrate. The sample spots on the ODT-coated plate were 100-fold smaller than those formed on an unmodified gold plate with a 1-mu l sample and generated 10 to 50 times higher mass sensitivity for peptide standards by MALDI-TOF MS. When the sample was deposited on an ODT-modified plate from a nanocolumn, the detection limit for peptides was as low as 20 pM for 5-mu l samples corresponding to 80 amol deposited. This technique was used to analyze extracts of microwave-fixed tissue from rat brain striatum. Ninety-eight putative peptides were detected including several that had masses matching neuropeptides expected in this brain region such as substance P, rimorphin, and neurotensin. Twenty-three peptides had masses that matched peaks detected by capillary liquid chromatography with electrospray ionization MS.(1,2) Copyright (c) 2005 John Wiley & Sons, Ltd.
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