Trace LC/MS quantitative analysis of polypeptide biomarkers: impact of 1-D and 2-D chromatography on matrix effects and sensitivity.
J Sep Sci
; 30(2): 226-33, 2007 Feb.
Article
em En
| MEDLINE
| ID: mdl-17390616
ABSTRACT
We investigated the impact of one dimension (single reverse phase (RP) column) and two dimension (two different RP columns) chromatographic methods on SIM (MS) and multiple reaction monitoring (MRM; MS/MS) performance from human plasma. We find that MRM analysis is clearly preferable for 1-D applications; however, implementation of SIM detection in conjunction with 2-D separation technique resulted in an over 60-fold increase in analyte peak area and improved S/N compared to MRM for our analyte, human C-peptide. Implementation of a 2-D RP-RP technique with SIM detection is capable of eliminating matrix effects and greatly increases signal response and data quality. For two large peptides in complex biological samples, we found that a 2-D approach performed better than high quality sample preparation together with 1-D chromatography and MRM, even on a high-end mass spectrometer.
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Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Espectrometria de Massas em Tandem
Idioma:
En
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
Estados Unidos