Capillary electrophoresis with laser-induced fluorescence detection for natively fluorescent analytes

Gooijer, C.; Kok, S.J.; Ariese, Freek

doi:10.1051/analusis:2000280679

Cited by 26 publications

(23 citation statements)

References 33 publications

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“…In the same vein, Yeung and coworkers proposed the use of a 257-nm Ar ion laser to detect conalbumin or insulin in the 100 pM range [17,18] and other authors have used pulsed laser for such measurements [19,20]. In addition, we note the remarkable work presenting the online recording of emission spectra for identification purposes, cryogenic interfaces for high-resolution spectroscopic identification, and the use of multiphoton excitation to perform UV-LIF which was presented by Gooijer et al [21] Acrylamide is believed to quench the fluorescence [22,23] of Trp when it does not react to form polyacrylamide [24], so we used polyethyleneoxide for the separation of a protein mixture, resulting in nM LOD [19]. Recently we proposed the use of an ellipsoidal LIF detector based on the work of Wan et al [25] which uses the fluorescence reflected inside the capillary and used a glued ellipsoid to collect this fluorescence.…”

Section: Introductionmentioning

confidence: 72%

A new evaluation technique for the detection of impurities in purified proteins via CE with native UV‐LIF

Rodat¹,

Couderc²,

Feurer³

et al. 2010

Electrophoresis

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An analytical methodology for quality control analyses of IgG and their impurities is presented using a new UV-LIF (266 nm) detector inside the cassette of a CE instrument and its performance was evaluated. The observed sensitivity was very close to that obtained by silver staining of slab gels (LOD of 25 ng/mL), while the sensitivity of the analysis is 80 times better than with CE/UV absorption (214 nm). Examples of the analysis of pharmaceutical and other commercial IgGs are provided and the kinetics of the reduction of IgG by beta-mercaptoethanol is reported, demonstrating the ease of performing the analysis.

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Section: Introductionmentioning

confidence: 72%

A new evaluation technique for the detection of impurities in purified proteins via CE with native UV‐LIF

Rodat¹,

Couderc²,

Feurer³

et al. 2010

Electrophoresis

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“…Within this range, gas lasers are the most commonly used sources: (i) He-Cd laser (325 and 442 nm), (ii) He-Ne laser (543.6, 592.6 and 633 nm), (iii) KrF excimer laser (248 nm), (iv) Nd-YAG (Nd-yttriumaluminium-garnet) laser (266 nm) and (v) Ar 1 ion laser (usually 488 and/or 514 nm) with powers ranging from a few milliwatts to more than 10 W [38]. Diode lasers are starting to be used with greater frequency as they are less expensive, highly compact, stable, thermoelectrically cooled and provide power of few milliwatts of quasi-continuous wave sufficient for LIF detection [39]. Wavelengths available with semiconductor diode lasers are 266, 355, 410, 488 and 532 nm.…”

Section: Instrumentation and Laser Sourcesmentioning

confidence: 99%

“…Wavelengths available with semiconductor diode lasers are 266, 355, 410, 488 and 532 nm. Extraspectral selectivity can be obtained by performing CE-LIF under fluorescence line-narrowing conditions using a cryogenic interface [39,40]. An alternative to deep UV-excitation is the use of an excitation process in which two or three visible or infrared photons act consecutively to bridge the energy gap between the S 0 and S 1 -electronic states of analytes.…”

Section: Instrumentation and Laser Sourcesmentioning

confidence: 99%

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LIF detection of peptides and proteins in CE

2007

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CE- and microchip-based separations coupled with LIF are powerful tools for the separation, detection and determination of biomolecules. CE with certain configurations has the potential to detect a small number of molecules or even a single molecule, thanks to the high spatial coherence of the laser source which permits the excitation of very small sample volumes with high efficiency. This review article discusses the use of LIF detection for the analysis of peptides and proteins in CE. The most common laser sources, basic instrumentation, derivatization modes and set-ups are briefly presented and special attention is paid to the different fluorogenic agents used for pre-, on- and postcapillary derivatization of the functional groups of these compounds. A table summarizing major applications of these derivatization reactions to the analysis of peptides and proteins in CE-LIF and a bibliography with 184 references are provided which covers papers published to the end of 2005.

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“…CE-LIFD systems make use of visible laser lines so that they can only be applied to analytes that have been chemically derivatized with a suitable fluorescent tag matching the excitation wavelength. But the CE-LIFD can be expanded to include natively fluorescent analytes by employing either improved UV pulse or continuous wave laser systems or multiphoton-excitation [164]. A highly sensitive LIFD detection system based on a 635-nm laser diode and a confocal microscope is used for planar microfluidic CE chip [165].…”

Section: Improved Detectionmentioning

confidence: 99%

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

Páez

Hernández

2001

Biopharm & Drug Disp

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Capillary electrophoresis (CE) is a high-efficiency analytical technique that has had a great impact as a tool in biomedical research, clinical and forensic practice in the last ten years. Only in one of the applications, the DNA analysis, it has had an explosive exponential growth in the last few years. This impact is expressed in an enormous amount of CE articles and many reviews. The CE advantages with respect to other analytical techniques: the required very small sample volume, rapid analysis, great resolution power and low costs, have made this technique ideal for the analysis of a numerous endogenous and exogenous substances present in biological fluids. The different modes of CE have been coupled to different detection techniques such as UV-absorbance, electrochemical, mass spectrometry and laser-induced fluorescence detection (LIFD) to detect different nature and molecular size separated analytes. This review focuses mostly on the applications of CE-LIFD, to measure drugs and endogenous neuroactive substances such as amino acids and monoamines, especially in microdialysis samples from experimental animals and humans. CE-LIFD trends are discussed: automated faster analysis with capillary array systems, resolution power improvement, higher detection sensitivity, and CE systems miniaturization for extremely small sample volume, in order to make CE easier and affordable to the lab bench or the clinical bed.

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Capillary electrophoresis with laser-induced fluorescence detection for natively fluorescent analytes

Cited by 26 publications

References 33 publications

A new evaluation technique for the detection of impurities in purified proteins via CE with native UV‐LIF

A new evaluation technique for the detection of impurities in purified proteins via CE with native UV‐LIF

LIF detection of peptides and proteins in CE

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

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