Document Detail


Qualitative and quantitative analysis of amino acids by capillary electrophoresis-electrospray ionization-tandem mass spectrometry.
MedLine Citation:
PMID:  15237395     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe a method to identify and quantify amino acids using capillary electrophoresis-electrospray ionization-triple-quadrupole tandem mass spectrometry (CE-ESI-MS/MS). Amino acids, including physiological amino acids, were first separated by CE under acidic pH conditions and then detected by MS/MS. To efficiently introduce the whole sample into the capillary, no electrical potential was applied to the electrospray probe until running electrophoresis. The position of the electrosprayer with respect to the MS capillary entrance drastically affected sensitivity and generation of cluster ions. MS/MS with multiple reaction monitoring (MRM) detection was performed to obtain sufficient selectivity and sensitivity. Under optimized CE-MS/MS conditions, the minimum detectable levels for 32 free amino acids normally found in proteins and other physiological amino acids were between 0.1 and 14 micromol/L with pressure injection of 50 mbar for 3 s (3 nL) at a signal-to-noise ratio of 3. For most amino acids, this constitutes a severalfold increase in sensitivity compared to CE-MS. The relative standard deviations (% RSD) for all amino acids were better than 0.4% for migration times and between 1.4% and 8.6% for peak areas (n = 10). Since amino acids exhibited characteristic MS/MS spectra, this approach is useful for the simultaneous, selective, quantitative, and reproducible analysis of amino acids in physiological and biological samples that contain various kinds of matrices. The power of the method was demonstrated by analyzing amino acids in human urine.
Authors:
Tomoyoshi Soga; Yuji Kakazu; Martin Robert; Masaru Tomita; Takaaki Nishioka
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Electrophoresis     Volume:  25     ISSN:  0173-0835     ISO Abbreviation:  Electrophoresis     Publication Date:  2004 Jul 
Date Detail:
Created Date:  2004-07-06     Completed Date:  2005-01-31     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8204476     Medline TA:  Electrophoresis     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1964-72     Citation Subset:  IM    
Affiliation:
Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata, Japan. soga@sfc.keio.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / urine*
Electrophoresis, Capillary / methods*
Humans
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization / methods*
Time Factors
Chemical
Reg. No./Substance:
0/Amino Acids

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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