High Efficiency Robust Open Tubular Capillary Electrochromatography Column for the Separation of Peptides

Ali, Faiz; Cheong, Won Jo

doi:10.1002/bkcs.10859

Cited by 8 publications

(20 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Scanning electron microscopy (SEM) has become an indispensable characterisation method for a visual confirmation of the surface morphology of coated surfaces. SEM images of successfully coated surfaces were described to show rough surfaces , hilly or bumpy surfaces , with distinct shapes of the embedded material , good layer density and homogeneity or uniformity , and more importantly significant layer thickness as opposed to the smooth surfaces seen in bare silica surfaces. Some SEM scans were performed on flat silica surfaces for detailed characterisation because of the inherent difficulty in successfully imaging wall coatings for a capillary format.…”

Section: Characterisation Methodsmentioning

confidence: 99%

“…These results reinforced previous findings from the same group using a shorter 60‐cm capillary coated with the same tri‐copolymer. This shorter column yielded the separation of about 20 peptide peaks from the tryptic digest sample of cytochrome C with high separation efficiency close to 500 000 plates/column and acceptable peak capacity of over 220 .…”

Section: Developments In Stationary Phase Materialsmentioning

confidence: 99%

“…The in situ synthesis and immobilisation of N‐phenylacrylamide‐styrene‐methacrylic acid copolymer layer onto the capillary by RAFT polymerization was developed . The reaction scheme started with reacting a pre‐treated capillary with 4‐(chloromethyl)phenyl isocyanate in the presence of dibutyltin dichloride as catalyst followed by sodium diethyl dithiocarbamate as the initiator moiety.…”

Section: Developments In Coating Strategies and Fabrication Techniquesmentioning

confidence: 99%

“…In the column prepared to separate cytochrome C tryptic digests, Ali and Cheong decreased levels of MAA to reduce very strong interactions of the SP with the analyte that caused band broadening . Toluene was employed as solvent in place of p‐xylene and included 4‐methyl‐2‐pentanone as a co‐solvent to improve compatibility of the polymerization mixture.…”

Section: Developments In Coating Strategies and Fabrication Techniquesmentioning

confidence: 99%

See 3 more Smart Citations

Recent developments in open tubular capillary electrochromatography from 2016 to 2017

2017

View full text Add to dashboard Cite

Recent developments in open tubular capillary electrochromatography from 2016 to 2017Interest in open-tubular capillary electrochromatography (OT-CEC) continues to thrive because of the inherent advantage of OT-CEC combining the high efficiency of capillary electrophoresis and the high selectivity of high performance liquid chromatography. For the period 2016 to 2017, novel materials have been developed as first-time stationary phases for OT-CEC and are grouped in this review as polymer-based materials, frameworks, nanoparticles, graphene-based materials, and biomaterials. Coating and fabrication methods mostly rely on covalent coating strategies while non-covalent immobilisation strategies like electrostatic assembly are notably still being employed. The concern of overcoming phase ratio challenges in OT-CEC coatings have also generated adoption of combined coating strategies including multi-layering, layer-by-layer self-assembly and methods adapted from nanofilm fabrications like epitaxial growth, liquid phase deposition, or nucleation of crystal growth. The emergence of non-conventional coating characterisation methods such as transmission electron microscopy, X-ray diffraction or X-ray photoelectron spectroscopy is also discussed.

show abstract

Section: Characterisation Methodsmentioning

confidence: 99%

Section: Developments In Stationary Phase Materialsmentioning

confidence: 99%

Section: Developments In Coating Strategies and Fabrication Techniquesmentioning

confidence: 99%

Section: Developments In Coating Strategies and Fabrication Techniquesmentioning

confidence: 99%

See 2 more Smart Citations

Recent developments in open tubular capillary electrochromatography from 2016 to 2017

2017

View full text Add to dashboard Cite

show abstract

“…13–16 Ali and Cheong reported a method for the immobilization of N-phenylacrylamide-styrene copolymer on the inner surface of capillaries with RAFT polymerization for applications in electrochromatograpy. 17–19 A ligand with a terminal halogen (4-chloromethylphenyl isocyanate), or 4-(trifluoromethoxy) phenyl isocyanate was bound to the inner surface of a pretreated silica capillary in the presence of dibutyltin dichloride as a catalyst through an isocyanate-hydroxyl reaction. 17–18 Attachment of initiator (sodium diethyl dithiocarbamate) to the bound ligand was carried out and followed by in situ polymerization.…”

Section: Introductionmentioning

confidence: 99%

Surface-Confined Aqueous Reversible Addition–Fragmentation Chain Transfer (SCARAFT) Polymerization Method for Preparation of Coated Capillary Leads to over 10 000 Peptides Identified from 25 ng HeLa Digest by Using Capillary Zone Electrophoresis-Tandem Mass Spectrometry

2017

View full text Add to dashboard Cite

A surface-confined aqueous reversible addition-fragmentation chain transfer (SCARAFT) polymerization method was developed to coat capillaries for use in capillary zone electrophoresis (CZE). SCARAFT polymerization primarily takes place on the inner surface of the capillary instead of in solution, which greatly improves the homogeneity of the coating. Capillaries treated with this coating produced an electroosmotic mobility of 2.8 ± 0.2 × 10−6 cm2·V−1·s−1 (N=3), which is roughly an order of magnitude lower than that of commercial linear polyacrylamide (LPA) coated capillaries. Coated capillaries were evaluated for bottom-up proteomic analysis using CZE. The very low electroosmotic mobility results in a 200-minute separation and improved single-shot analysis. An average of 977 protein groups and 5,605 unique peptides were identified from 50 ng of an E. coli digest and 2,158 protein groups and 10,005 peptides were identified from 25 ng of a HeLa digest using single-shot analysis with a SCARAFT-acrylamide capillary coupled to a Q Exactive HF mass spectrometer. The coating is stable. A single capillary was used for over 200 hours (8.4 days) of continuous operation. RSD in migration time was between 2 and 3% for selected ion electropherograms (SIEs) generated for six ions; median theoretical plate counts ranged from 240,000 to 600,000 for these SIEs. Various types of coatings could be prepared by simply changing the functional vinyl monomers in the polymerization mixture. Positively charged coatings using direct attachment and formation of a block-copolymer were prepared and demonstrated for the separation of mixtures of intact proteins.

show abstract

Open tubular capillary column of 50 μm internal diameter with a very high separation efficiency for the separation of peptides in CEC and LC

Ali

Cheong

2017

J of Separation Science

Self Cite

View full text Add to dashboard Cite

A specially designed long open tubular capillary column (50 μm internal diameter and 112 cm effective length) was prepared by fabrication of a thin three-component co-polymer layer on the inner surface of silica capillary. A pretreated silica capillary was reacted with 4-(chloromethyl)phenyl isocyanate in the presence of dibutyltin dichloride as catalyst followed by sodium diethyl dithiocarbamate. Then a thin polymer layer was made on the inner surface of capillary by reversible addition-fragmentation transfer polymerization of styrene, N-phenylacrylamide, and methacrylic acid. A carefully adjusted formulation of reaction mixture and elaborated procedures were adopted to secure formation of the co-polymer layer of enhanced separation performance. The co-polymer immobilized open tubular capillary column was used for the separation of a synthetic mixture of five peptides and excellent separation efficiency (over 1.7 million per column) was obtained in the capillary electrochromatography mode. Such excellent separation efficiencies of ca. 1 m column have not been obtained in the isocratic elution mode so far. The column was also used for separation of the peptides in the liquid chromatography mode to show very good separation efficiency (average 286 700 per column).

show abstract

High Efficiency Robust Open Tubular Capillary Electrochromatography Column for the Separation of Peptides

Cited by 8 publications

References 21 publications

Recent developments in open tubular capillary electrochromatography from 2016 to 2017

Recent developments in open tubular capillary electrochromatography from 2016 to 2017

Surface-Confined Aqueous Reversible Addition–Fragmentation Chain Transfer (SCARAFT) Polymerization Method for Preparation of Coated Capillary Leads to over 10 000 Peptides Identified from 25 ng HeLa Digest by Using Capillary Zone Electrophoresis-Tandem Mass Spectrometry

Open tubular capillary column of 50 μm internal diameter with a very high separation efficiency for the separation of peptides in CEC and LC

Contact Info

Product

Resources

About