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In situ liquid-liquid extraction as a sample preparation method for matrix-assisted laser desorption/ionization MS analysis of polypeptide mixtures

Jensen, Ole Nørregaard and Kjellström, Sven (2003) In Analytical Chemistry 75(10). p.9-2362
Abstract

A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic and hydrophilic polypeptides in mixtures, thereby reducing the complexity of mass spectra obtained by MALDI MS. The LLE technique was optimized for rapid and sensitive in situ (on-target) sample preparation for MALDI MS analysis of proteins and peptides at low-picomole and subpicomole levels. Addition of MALDI matrix to the organic solvent enhanced the efficiency of the LLE-MALDI MS method for analysis of hydrophobic peptides and proteins. LLE-MALDI MS... (More)

A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic and hydrophilic polypeptides in mixtures, thereby reducing the complexity of mass spectra obtained by MALDI MS. The LLE technique was optimized for rapid and sensitive in situ (on-target) sample preparation for MALDI MS analysis of proteins and peptides at low-picomole and subpicomole levels. Addition of MALDI matrix to the organic solvent enhanced the efficiency of the LLE-MALDI MS method for analysis of hydrophobic peptides and proteins. LLE-MALDI MS enabled the detection of the hydrophobic membrane protein bacteriorhodopsin as a component in a simple protein mixture. Peptide mixtures containing phosphorylated, glycosylated, or acylated peptides were successfully separated and analyzed by the in situ LLE-MALDI MS technique and demonstrate the potential of this method for enhanced separation and structural analysis of posttranslationally modified peptides in proteomics research.

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author
publishing date
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Contribution to journal
publication status
published
subject
keywords
Amino Acid Sequence, Animals, Chemical Fractionation, Peptides, Phosphopeptides, Sensitivity and Specificity, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Journal Article, Research Support, Non-U.S. Gov't
in
Analytical Chemistry
volume
75
issue
10
pages
8 pages
publisher
The American Chemical Society
external identifiers
  • scopus:0038290681
ISSN
0003-2700
DOI
10.1021/ac026297w
language
English
LU publication?
no
id
4c89eee0-72c2-4466-ad50-e3f34634dfc8
date added to LUP
2016-12-13 14:43:25
date last changed
2017-03-17 18:12:01
@article{4c89eee0-72c2-4466-ad50-e3f34634dfc8,
  abstract     = {<p>A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic and hydrophilic polypeptides in mixtures, thereby reducing the complexity of mass spectra obtained by MALDI MS. The LLE technique was optimized for rapid and sensitive in situ (on-target) sample preparation for MALDI MS analysis of proteins and peptides at low-picomole and subpicomole levels. Addition of MALDI matrix to the organic solvent enhanced the efficiency of the LLE-MALDI MS method for analysis of hydrophobic peptides and proteins. LLE-MALDI MS enabled the detection of the hydrophobic membrane protein bacteriorhodopsin as a component in a simple protein mixture. Peptide mixtures containing phosphorylated, glycosylated, or acylated peptides were successfully separated and analyzed by the in situ LLE-MALDI MS technique and demonstrate the potential of this method for enhanced separation and structural analysis of posttranslationally modified peptides in proteomics research.</p>},
  author       = {Jensen, Ole Nørregaard and Kjellström, Sven},
  issn         = {0003-2700},
  keyword      = {Amino Acid Sequence,Animals,Chemical Fractionation,Peptides,Phosphopeptides,Sensitivity and Specificity,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  month        = {05},
  number       = {10},
  pages        = {9--2362},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {In situ liquid-liquid extraction as a sample preparation method for matrix-assisted laser desorption/ionization MS analysis of polypeptide mixtures},
  url          = {http://dx.doi.org/10.1021/ac026297w},
  volume       = {75},
  year         = {2003},
}