Capillary electrochromatography of biomolecules with on-line electrospray ionization and time-of-flight mass spectrometry

Choudhary, Gargi; Horváth, Csaba; Banks, J. Fred

doi:10.1016/s0021-9673(98)00817-6

Cited by 66 publications

(38 citation statements)

References 44 publications

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“…After the polyimide coating of the column end is burned off, the capillary end is moved quickly in and out of the flame center until the taper is gradually formed over a period of 30-60 s. To prevent deterioration of tapered end, the capillary is rotated during the heating process. Compared to the published fabrication method [24], our procedure does not allow the taper to close the internal channel completely. Hence, the internal tapered capillary with id 7-10 mm can be produced reproducibly without further sanding.…”

Section: Cec Column Fabricationmentioning

confidence: 99%

Simultaneous enantioseparation and sensitive detection of eight β‐blockers using capillary electrochromatography‐electrospray ionization‐mass spectrometry

Zheng

Shamsi

2006

Electrophoresis

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The feasibility of using vancomycin chiral stationary phase (CSP) and polar organic eluent is investigated for simultaneous enantioseparation of eight beta-blockers using CEC coupled to ESI mass spectrometric detection (ESI-MS). The internally tapered capillaries were utilized to pack CEC-MS columns. As compared to externally tapered columns, the use of internally tapered columns demonstrated enhanced stability, durability, and reproducibility. A mixture containing methanol/ACN/acetic acid/triethylamine at 70:30:1.6:0.2 v/v/v/v was considered as optimum mobile phase since it provided a good compromise between resolution and analysis time. As expected, sheath liquid and ESI-MS parameters mainly influenced the detection sensitivity. Interestingly, structural information of beta-blockers was available by varying the MS fragmentor voltage using in-house CID in the scan mode. In order to maximize the chiral/achiral resolution, various column-coupling approaches using teicoplanin as complementary CSP to vancomycin were tested. Several changes in the elution order of beta-blockers were observed using multimodal CSPs with some improvement in chiral or achiral resolution. The quantitative aspects of the CEC-MS method were demonstrated using R- and S-talinolol as internal standards. The calibration curves of beta-blockers showed good linearity in the range of 3-600 microM. The enantiomer of beta-blockers at a concentration of 30 nM was detectable. Furthermore, both 0.1 and 1% of the S-enantiomer could be precisely quantified in the presence of 99.9 and 99% of the R-isomer of beta-blocker.

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Section: Cec Column Fabricationmentioning

confidence: 99%

Simultaneous enantioseparation and sensitive detection of eight β‐blockers using capillary electrochromatography‐electrospray ionization‐mass spectrometry

Zheng

Shamsi

2006

Electrophoresis

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“…Additionally, the usage of in-line filters to keep the stationary phase from leaving the column also leads to the limitation of permeability and mechanical strength, increasing the extra-column band broadening processes. Over the past decades, different strategies have been developed to prepare a frit for retention of the stationary phase in capillaries such as using the taper end columns, [9][10][11][12][13] coupled capillaries via a polytetrafluoroethylene (PTFE) sleeve or restrictors, 14,15 silica-based sintered frits, [16][17][18] monolithic frits, etc. [19][20][21][22] In this paper, a new approach using sintered stainless steel powders as an inlet frit was firstly introduced for the manufacture of particulate packed capillary columns in our laboratory.…”

Section: Introductionmentioning

confidence: 99%

A Simple Design to Realize Micro‐column Separation by Conventional Analytical HPLC

Gong

Zhang

Zhang³

et al. 2009

Chin. J. Chem.

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The conventional analytical HPLC was successfully developed for micro-column separation by using a simple eluate splitting system, self-preparation of packing column and on-capillary column detector in our laboratory. Porous inlet frit in fused silica capillary was rapidly prepared by sintering stainless steel powders under 500 meshes for about 20 s. The use of such frits or metal meshes in capillary to retain C 18 particles of chromatographic packing was demonstrated to be stable and specially robust with continuous packing and long chromatographic runs. Furthermore, the chromatographic behavior was detailedly evaluated by changing the flow rate and the percentage of mobile phase using the prepared capillary column. Under the optimal experimental conditions, baseline separation of the model analytes including thiourea, benzene, toluene, ethylbenzene was obtained with a high column efficiency near 70000N (plates/m) by the developed capillary-HPLC.

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“…Although no frit is needed in open tubular (OT) and monolithic CEC, OT-CEC has limited sample capacity, and a trial and error optimization is often needed to select the composition of the polymerization in monolithic CEC. To take advantage of the large variety of commercial stationary phases for HPLC, packed CEC without the use of frit has been proposed [6,[14][15][16][17][18][19][20]. For example, Lord et al [19] have reported the use of a tapered tip instead of frit at the outlet of a packed CEC column.…”

Section: Introductionmentioning

confidence: 99%

Analysis of triazines by capillary electrochromatography/electrospray ionization‐mass spectrometry using a low‐flow sheath liquid interface

et al. 2006

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CEC-MS has been used for the analysis of eight-triazine herbicides. It showed significantly better S/N ratio than reversed EOF CE-MS and MEKC-MS, due to the lack of a surfactant in the separation buffer. By optimizing the pH, the organic content of the running buffer, and the separation potential, optimal separation was achieved within 18 min using a running buffer of pH 7.0, containing 70% v/v ACN, and an applied voltage of 17 kV. Gradient CEC showed superior separation when compared with isocratic elution. The combination of a tapered CEC column and a low-flow interface confers several advantages including better sensitivity, low dead volume, and independent control of the conditions used for CEC separation and ESI analysis.

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Capillary electrochromatography of biomolecules with on-line electrospray ionization and time-of-flight mass spectrometry

Cited by 66 publications

References 44 publications

Simultaneous enantioseparation and sensitive detection of eight β‐blockers using capillary electrochromatography‐electrospray ionization‐mass spectrometry

Simultaneous enantioseparation and sensitive detection of eight β‐blockers using capillary electrochromatography‐electrospray ionization‐mass spectrometry

A Simple Design to Realize Micro‐column Separation by Conventional Analytical HPLC

Analysis of triazines by capillary electrochromatography/electrospray ionization‐mass spectrometry using a low‐flow sheath liquid interface

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