Use of a Mixed-Mode Packing and Voltage Tuning for Peptide Mixture Separation in Pressurized Capillary Electrochromatography with an Ion Trap Storage/Reflectron Time-of-Flight Mass Spectrometer Detector

Huang, Peiqing; Jin, Xiaoying; Chen, Yajuan; Srinivasan, Jannavi; Lubman, David M.

doi:10.1021/ac980813u

Cited by 94 publications

(39 citation statements)

References 22 publications

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“…This causes interferences with the detection of the analyte of interest. Thus far, several studies have been performed using packed-CEC/MS [198][199][200][201][202][203][204]. In contrast, only a few studies have been reported on OT-CEC/MS [18,101,121,205].…”

Section: Coupling With Mass Spectrometrymentioning

confidence: 99%

Analytical separations in open‐tubular capillary electrochromatography

Kapnissi‐Christodoulou

Zhu

Warner

2003

Electrophoresis

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This review represents a summary of recent progress in open-tubular capillary electrochromatography (OT-CEC) for chiral and achiral separations. The OT-CEC approach is an alternative to packed-CEC that could eliminate the problems associated with retaining frits and silica particles. In OT-CEC, the stationary phase is immobilized on the inner walls of the capillary. Preparation of the stationary phase is critical for OT-CEC. The preparation methods for capillary columns include (i) adsorption, (ii) covalent bonding and/or cross-linking, (iii) porous layers, (iv) chemical bonding after etching, (v) sol-gel, and (vi) molecular imprinting. Major developments, potential applications, technical difficulties and advantages associated with these wall coatings in OT-CEC are presented. In addition, the coupling of OT-CEC with mass spectrometry (MS) is briefly reviewed. Several applications of this hyphenated technique for analytical separations are also summarized.

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Section: Coupling With Mass Spectrometrymentioning

confidence: 99%

Analytical separations in open‐tubular capillary electrochromatography

Kapnissi‐Christodoulou

Zhu

Warner

2003

Electrophoresis

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“…The concomitant use of pressure assistance increases the mobile phase flow rate to take advantage of the enhanced mass-transfer of the monolithic stationary phases. The influence of pressure in electrically driven chromatography has been investigated by several groups [9,23,35,36]. The combination of the electrophoretic flat flow profile and the pressure-driven parabolic Poiseuille flow profile results in less peak broadening than pressure-driven flow alone.…”

Section: Effect Of Pressure Drop and Buffer Ph On Separation Performancementioning

confidence: 99%

“…The combination of the electrophoretic flat flow profile and the pressure-driven parabolic Poiseuille flow profile results in less peak broadening than pressure-driven flow alone. Tuning of peptide elution by adjustment of the electric field and the inlet pressure has been demonstrated with use of particulate columns [35].…”

Section: Effect Of Pressure Drop and Buffer Ph On Separation Performancementioning

confidence: 99%

High‐efficiency peptide analysis on monolithic multimode capillary columns: Pressure‐assisted capillary electrochromatography/capillary electrophoresis coupled to UV and electrospray ionization‐mass spectrometry

2003

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High-efficiency peptide analysis using multimode pressure-assisted capillary electrochromatography/capillary electrophoresis (pCEC/pCE) monolithic polymeric columns and the separation of model peptide mixtures and protein digests by isocratic and gradient elution under an applied electric field with UV and electrospray ionization-mass spectrometry (ESI-MS) detection is demonstrated. Capillary multipurpose columns were prepared in silanized fused-silica capillaries of 50, 75, and 100 microm inner diameters by thermally induced in situ copolymerization of methacrylic monomers in the presence of n-propanol and formamide as porogens and azobisisobutyronitrile as initiator. N-Ethylbutylamine was used to modify the chromatographic surface of the monolith from neutral to cationic. Monolithic columns were termed as multipurpose or multimode columns because they showed mixed modes of separation mechanisms under different conditions. Anion-exchange separation ability in the liquid chromatography (LC) mode can be determined by the cationic chromatographic surface of the monolith. At acidic pH and high voltage across the column, the monolithic stationary phase provided conditions for predominantly capillary electrophoretic migration of peptides. At basic pH and electric field across the column, enhanced chromatographic retention of peptides on monolithic capillary column made CEC mechanisms of migration responsible for separation. The role of pressure, ionic strength, pH, and organic content of the mobile phase on chromatographic performance was investigated. High efficiencies (exceeding 300 000 plates/m) of the monolithic columns for peptide separations are shown using volatile and nonvolatile, acidic and basic buffers. Good reproducibility and robustness of isocratic and gradient elution pressure-assisted CEC/CE separations were achieved for both UV and ESI-MS detection. Manipulation of the electric field and gradient conditions allowed high-throughput analysis of complex peptide mixtures. A simple design of sheathless electrospray emitter provided effective and robust low dead volume interfacing of monolithic multimode columns with ESI-MS. Gradient elution pressure-assisted mixed-mode separation CE/CEC-ESI-MS mass fingerprinting and data-dependent pCE/pCEC-ESI-MS/MS analysis of a bovine serum albumin (BSA) tryptic digest in less than 5 min yielding high sequence coverage (73%) demonstrated the potential of the method.

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“…Lubman and co-workers [146,147] used C 18 or mixed mode packing in the pressurized capillary electrochromatography (pCEC) mode for the peptides analysis. The pCEC affords faster separation of peptides with enhanced resolution, compared to LC.…”

Section: Silica Stationary Phasesmentioning

confidence: 99%

“…Conditions: mobile-phase A, 0.025% TFA B, 0.025% TFA in ACN; gradient of 0-100% B in 40 min; voltage, 1700 V; pressure, 80 bar; columns (a) 85 mm6150 mm packed with C18 reversed-phase and (b) 85 mm6150 mm packed with mixed-mode phase. Reprinted from [147], with permission. demonstrated the potential of the hyphenation CEC-MS for peptide analyses via a nanoelectrospray interface.…”

Section: Silica Stationary Phasesmentioning

confidence: 99%

Recent progress in high‐performance capillary bioseparations

et al. 2003

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Bioanalysis is a fast growing domain with new technological developments in the field of mass spectrometry, separation science, and bioinformatics. The central role of separation prior to detection and evaluation of analytes is indisputable. In fact, since the decipherment of the human genome via multichannel capillary electrophoresis in combination with laser-induced fluorescence [1], the analysis of the proteome, i.e., the type and amount of all proteins expressed at a certain time by the genome in a cell or tissue [2] became of central interest. Therefore, new fast and sensitive capillary liquid chromatographic, electrophoretic, and electrochromatographic separation techniques, suitable for high-throughput analysis of proteins, peptides and nucleic acids, are required.

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Use of a Mixed-Mode Packing and Voltage Tuning for Peptide Mixture Separation in Pressurized Capillary Electrochromatography with an Ion Trap Storage/Reflectron Time-of-Flight Mass Spectrometer Detector

Cited by 94 publications

References 22 publications

Analytical separations in open‐tubular capillary electrochromatography

Analytical separations in open‐tubular capillary electrochromatography

High‐efficiency peptide analysis on monolithic multimode capillary columns: Pressure‐assisted capillary electrochromatography/capillary electrophoresis coupled to UV and electrospray ionization‐mass spectrometry

Recent progress in high‐performance capillary bioseparations

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