title: Trace LC/MS quantitative analysis of polypeptide biomarkers: impact of 1-D and 2-D chromatography on matrix effects and sensitivity.
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.
authors: Core Analytical Laboratory at General Clinical Research Center, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY 10461, USA. rogatsky@aecom.yu.edu
authors: Eduard
authors: E
authors: Rogatsky
authors: Vlad
authors: V
authors: Tomuta
authors: Harsha
authors: H
authors: Jayatillake
authors: Greg
authors: G
authors: Cruikshank
authors: Louis
authors: L
authors: Vele
authors: Daniel T
authors: DT
authors: Stein
mesh_terms: D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods; D005934:Glucagon / Q000737:chemistry; D006801:Humans; D010455:Peptides / Q000032:analysis* / Q000737:chemistry*; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000295:instrumentation* / Q000379:methods*
chemical_list: D015415:Biomarkers; D010455:Peptides; D005934:Glucagon