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A Separation Voltage Polarity Switching Method for Higher Sample Loading Capacity and Better Separation Resolution in Transient Capillary Isotachophoresis Separation

Journal Article · · Analyst
DOI:https://doi.org/10.1039/C8AN01779D· OSTI ID:1494984
 [1];  [2];  [2];  [2];  [2]
  1. UNIVERSITY PROGRAMS
  2. BATTELLE (PACIFIC NW LAB)
+ Show Author Affiliations

A separation voltage polarity switching transient capillary isotachophoresis (PS-tCITP) was developed to overcome a major sample loading volume limitation in transient capillary isotachophoresis (tCITP). The fundamental idea of PS-tCITP is to let sample ions move back and forth in the separation capillary during their initial isotachophoresis focusing stage by switching the polarity of separation voltage, in order to both increase the sample loading volume and improve the separation efficiency as compared to the conventional tCITP method. The experimental evaluation of the novel PS-tCITP method by using two peptide standards at 2 µM concentration showed that the maximum sample loading volume could be increased from 45% of the total separation capillary volume in tCITP to 70% in PS-tCITP, which resulted in a more than 1.5 fold increases in the peptide peak intensity at a given length/volume of the separation capillary. Due to the consecutive focusing of sample volume from each polarity switching of separation voltage, the separation time window at a given sample loading volume was also increased significantly in PS-tCITP as compared to tCITP. Experiment comparison between tCITP and PS-tCITP at 45% sample loading volume using the same setup showed that the migration time difference between the two peptide peaks increased from 0.3 min in tCITP to 0.363 min in PS-tCITP with similar peak widths and heights, resulting in roughly a 21% improvement in separation resolution. The performance advantages of PS-tCITP separation over tCITP separation was further verified by using a mixture of six peptide standards.

Research Organization:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
AC05-76RL01830
OSTI ID:
1494984
Report Number(s):
PNNL-SA-135251; ANALAO
Journal Information:
Analyst, Vol. 144, Issue 2; ISSN 0003-2654
Publisher:
Royal Society of Chemistry
Country of Publication:
United States
Language:
English

References (32)

Simplified capillary electrophoresis nanospray sheath-flow interface for high efficiency and sensitive peptide analysis: Capillary electrophoresis electrospray interface July 2010
Over 2300 Phosphorylated Peptide Identifications with Single-Shot Capillary Zone Electrophoresis-Tandem Mass Spectrometry in a 100 min Separation September 2015
In Situ Microprobe Single-Cell Capillary Electrophoresis Mass Spectrometry: Metabolic Reorganization in Single Differentiating Cells in the Live Vertebrate ( Xenopus laevis ) Embryo May 2017
Capillary Isotachophoresis-Nanoelectrospray Ionization-Selected Reaction Monitoring MS via a Novel Sheathless Interface for High Sensitivity Sample Quantification July 2013
Capillary Electrophoresis–Nanoelectrospray Ionization–Selected Reaction Monitoring Mass Spectrometry via a True Sheathless Metal-Coated Emitter Interface for Robust and High-Sensitivity Sample Quantification April 2016
Optimization and Evaluation of a Sheathless Capillary Electrophoresis–Electrospray Ionization Mass Spectrometry Platform for Peptide Analysis: Comparison to Liquid Chromatography–Electrospray Ionization Mass Spectrometry October 2011
Bottom-Up Proteomics of Escherichia coli Using Dynamic pH Junction Preconcentration and Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry June 2014
Novel electrophoretic acetonitrile-based stacking for sensitive monitoring of the antiepileptic drug perampanel in human serum October 2018
Ultrasensitive Sample Quantitation via Selected Reaction Monitoring Using CITP/CZE–ESI-Triple Quadrupole MS November 2012
Simplifying CE−MS Operation. 2. Interfacing Low-Flow Separation Techniques to Mass Spectrometry Using a Porous Tip June 2007
Multiple capillary isotachophoresis with repetitive hydrodynamic injections for performance improvement of the electromigration preconcentration July 2016
Transient isotachophoresis in on-line solid phase extraction capillary electrophoresis time-of-flight-mass spectrometry for peptide analysis in human plasma June 2011
Decoupling CE and ESI for a more robust interface with MS April 2010
Over 10 000 Peptide Identifications from the HeLa Proteome by Using Single-Shot Capillary Zone Electrophoresis Combined with Tandem Mass Spectrometry October 2014
Ultrasensitive and Fast Bottom-up Analysis of Femtogram Amounts of Complex Proteome Digests October 2013
Recent developments in capillary and microchip electroseparations of peptides (2015-mid 2017) September 2017
Enhancing the Coverage of the Urinary Metabolome by Sheathless Capillary Electrophoresis-Mass Spectrometry December 2011
High Capacity Capillary Electrophoresis-Electrospray Ionization Mass Spectrometry: Coupling a Porous Sheathless Interface with Transient-Isotachophoresis November 2010
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity January 2012
Application of capillary isotachophoresis in peptide analysis September 1991
Single-Shot Proteomics Using Capillary Zone Electrophoresis–Electrospray Ionization-Tandem Mass Spectrometry with Production of More than 1 250 Escherichia coli Peptide Identifications in a 50 min Separation February 2013
Third-Generation Electrokinetically Pumped Sheath-Flow Nanospray Interface with Improved Stability and Sensitivity for Automated Capillary Zone Electrophoresis–Mass Spectrometry Analysis of Complex Proteome Digests March 2015
Capillary zone electrophoresis–mass spectrometry with microliter-scale loading capacity, 140 min separation window and high peak capacity for bottom-up proteomics January 2017
Selective enrichment and ultrasensitive identification of trace peptides in proteome analysis using transient capillary isotachophoresis/zone electrophoresis coupled with nano-ESI-MS September 2006
Recent advances in enrichment techniques for trace analysis in capillary electrophoresis: CE and CEC September 2012
Recent progress of sample stacking in capillary electrophoresis (2014-2016): CE and CEC August 2016
Single-Shot Top-Down Proteomics with Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry for Identification of Nearly 600 Escherichia coli Proteoforms November 2017
Coupling porous sheathless interface MS with transient-ITP in neutral capillaries for improved sensitivity in glycopeptide analysis: CE and CEC January 2013
Simple Capillary Electrophoresis–Mass Spectrometry Method for Complex Glycan Analysis Using a Flow-Through Microvial Interface June 2014
Online Preconcentration in Capillaries by Multiple Large-Volume Sample Stacking: An Alternative to Immunoassays for Quantification of Amyloid Beta Peptides Biomarkers in Cerebrospinal Fluid January 2018
Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2014-2016): CE and CEC November 2016
Capillary Electrophoresis with Electrospray Ionization Mass Spectrometric Detection for Single-Cell Metabolomics July 2009

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