High resolution multichannel fluorescence detection for capillary electrophoresis Application to multicomponent analysis

Oldenburg, Kurt E.; Xi, Xiaoyan; Sweedler, Jonathan V.

doi:10.1016/s0021-9673(97)00713-9

Cited by 11 publications

(10 citation statements)

References 27 publications

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“…The optical detection scheme has been described in detail previously. 24 For this application, the 351-356 nm output of an argon-krypton ion laser (Innova Spectrum 70; Coherent, Palo Alto, CA, USA) is used for fluorescence excitation. The beam is focused into the 3 mm id quartz flow cell 5 mm beneath the capillary outlet using a 30 mm focal length fused-silica planoconvex lens (Spindler and Hoyer, Milford, MA, USA).…”

Section: Capillary Electrophoresis Systemmentioning

confidence: 99%

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Simple Sheath Flow Reactor for Post-column Fluorescence Derivatization in Capillary Electrophoresis

Oldenburg¹,

Sweedler²

1997

Analyst

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A system for post-column fluorescence derivatization in capillary electrophoresis is described. The post-column reactor uses a sheath flow detection cell where the reagents, o-phthaldialdehyde and beta-mercaptoethanol, are added to the sheath buffer and mix by diffusion with the analytes effusing from the separation capillary. Reaction progress is monitored and optimized by imaging a large portion of the sheath flow cuvette using an extended UV source and a CCD camera. Significantly, this design provides the ability to switch between the analysis of pre- and post-column derivatized amino acids and peptides easily and without sacrificing system performance. The lack of turbulent flow in this system minimizes post-separation band broadening. The limit of detection for glycine is 9.4 x 10(-8) M (110 amol) with a separation efficiency of 190,000 theoretical plates, without stacking. The performance of the system for a series of amino acids was evaluated using post-column and pre-capillary derivatization.

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Section: Capillary Electrophoresis Systemmentioning

confidence: 99%

“…The system uses the same flow assembly as used in previous visible and UV fluorescence experiments without modification. 6,24 Thus, we can switch from pre-to postcapillary derivatization by changing only the sheath buffer composition, providing a flexible alternative to specialized separation instrumentation.…”

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confidence: 99%

Simple Sheath Flow Reactor for Post-column Fluorescence Derivatization in Capillary Electrophoresis

Oldenburg¹,

Sweedler²

1997

Analyst

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“…The high irradiance and directionality of lasers are advantageous for focussing light into the small detection volume inside the capillary. CE-WRFlu has primarily been used for the analysis of native-fluorescent or fluorescently-labelled low-molecular-weight compounds such as dyes [6,7], metabolites [8][9][10], neurotransmitters [11][12][13][14][15][16][17], polyaromatic compounds [18,19], collagen cross linkers [20] and isoflavanoids [21]. Timperman et al [22,23] have analysed tyrosine and Trp-containing peptides, and Fultz et al [24] have measured dye-labelled proteins with LIF-based CE-WRFlu.…”

Section: Introductionmentioning

confidence: 99%

Lamp‐based wavelength‐resolved fluorescence detection for protein capillary electrophoresis: Setup and detector performance

2010

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A lamp-based fluorescence detection (Flu) system for CE was extended with a wavelength-resolved (WR) detector to allow recording of full protein emission spectra. WRFlu was achieved using a fluorescence cell that employs optical fibres to lead excitation light from a Xe-Hg lamp to the capillary window and protein fluorescence emission to a spectrograph equipped with a CCD. A 280 nm band pass filter etc. together with a 300 nm short pass cut-off filter was used for excitation. A capillary cartridge was modified to hold the detection cell in a commercial CE instrument enabling WRFlu in routine CE. The performance of the WRFlu detection was evaluated and optimised using lysozyme as model protein. Based on reference spectral data, a signal-intensity adjustment was introduced to correct for transmission losses in the detector optics that occurred for lower protein emission wavelengths. CE-WRFlu of lysozyme was performed using BGEs of 50 mM sodium phosphate (pH 6.5 or 3.0) and a charged-polymer coated capillary. Using the 3-D data set, signal averaging over time and emission-wavelength intervals was carried out to improve the S/N of emission spectra and electropherograms. The detection limit for lysozyme was 21 nM, providing sufficient sensitivity to obtain spectral information on protein impurities.

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“…[3,9,10]) or fiber-coupled systems [12]. CCD cameras [6,8,22] and charge injection device (CID) cameras [23,24] are being used because, with their two-dimensional array of pixels, they offer the possibility of simultaneously distinguishing between the different capillaries in one dimension (spatial resolution, allowing a true parallel detection) and using the other dimension for discriminating between the different fluorescent dyes (spectral resolution). When using off-column detection, the CCD camera can also be used to view twodimensional arrays of capillaries [13].…”

Section: Detectionmentioning

confidence: 99%

A fully automated multicapillary electrophoresis device for DNA analysis

et al. 1999

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High resolution multichannel fluorescence detection for capillary electrophoresis Application to multicomponent analysis

Cited by 11 publications

References 27 publications

Simple Sheath Flow Reactor for Post-column Fluorescence Derivatization in Capillary Electrophoresis

Simple Sheath Flow Reactor for Post-column Fluorescence Derivatization in Capillary Electrophoresis

Lamp‐based wavelength‐resolved fluorescence detection for protein capillary electrophoresis: Setup and detector performance

A fully automated multicapillary electrophoresis device for DNA analysis

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