Development of a novel fluorescent tag O-2-[aminoethyl]fluorescein for the electrophoretic separation of oligosaccharides

Kazarian, Artaches A.; Smith, Jason A.; Hilder, Emily F.; Breadmore, Michael C.; Quirino, Joselito P.; Suttil, James

doi:10.1016/j.aca.2010.01.011

Cited by 14 publications

(8 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Improvements of 515 over a conventional system were obtained. Based on this work, an analogous fluorescent reagent was synthesized [68] and used for the first demonstration of dynamic pH junction on a microchip [69]. The microchip was designed with an off-set cross defining a sample volume of 7% of the separation channel, the length that was found optimal in CE experiments and the derivatised sugars were separated within 120 s.…”

Section: Best Sensitivity Enhancements and Lodsmentioning

confidence: 99%

Online sample pre‐concentration via dynamic pH junction in capillary and microchip electrophoresis

Kazarian

Hilder

Breadmore

2011

J of Separation Science

Self Cite

View full text Add to dashboard Cite

Various analytical techniques have been developed over the years to analyse a large diversity of biomolecules with a constant push towards ultra-sensitive detection. CE is at the forefront of the most powerful analytical tools available to date when considering its superior efficiency and resolution; however, the technique suffers from poor sensitivity as a result of the short path length at the detection site and small injection volumes (typically <1% capillary length). One of the approaches to abate the inherent problem is to employ clever chemistry using sample focusing techniques whereby a large sample plug can be injected, preconcentrated and separated, producing excellent sensitivity and efficiency at the detector. This particular review will focus on the use of dynamic pH junction as a means of improving sensitivity in CE and focuses on the use of a change in analyte ionisation due to different pHs between the sample and electrolyte. The review provides a fundamental discussion of the mechanisms, buffer and sample conditions required to concentrate various analytes and a comprehensive list of published works in tabular format for easy identification of suitable conditions for new applications. The review further encompasses the use of dynamic pH junction in CE and its involvement in combination with other preconcentrations techniques to produce high sensitivity enhancements recorded between the years 1990-2010.

show abstract

Section: Best Sensitivity Enhancements and Lodsmentioning

confidence: 99%

Online sample pre‐concentration via dynamic pH junction in capillary and microchip electrophoresis

Kazarian

Hilder

Breadmore

2011

J of Separation Science

Self Cite

View full text Add to dashboard Cite

show abstract

“…Kazarian et al used dynamic pH junction in a glass/PDMS microchip to improve the sensitivity of sugars derivatised with a novel fluorescein derivative . A double T design was used in combination with polyelectrolyte multilayer coatings to provide a pH independent EOF to allow loading of the low pH sample.…”

Section: Stackingmentioning

confidence: 99%

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2010–2012)

et al. 2013

Self Cite

View full text Add to dashboard Cite

CE has been alive for over two decades now, yet its sensitivity is still regarded as being inferior to that of more traditional methods of separation such as HPLC. As such, it is unsurprising that overcoming this issue still generates much scientific interest. This review continues to update this series of reviews, first published in Electrophoresis in 2007, with updates published in 2009 and 2011 and covers material published through to June 2012. It includes developments in the field of stacking, covering all methods from field amplified sample stacking and large volume sample stacking, through to isotachophoresis, dynamic pH junction and sweeping. Attention is also given to online or inline extraction methods that have been used for electrophoresis.

show abstract

“…Polydimethylsiloxane (PDMS) was obtained from Sylgard, Dow Corning Corporation (Midland, MI). O-2-[aminoethyl]fluorescein was synthesised in-house as per Kazarian et al 47…”

Section: Chemicals and Materialsmentioning

confidence: 99%

“…This led us to synthesise a new ionisable fluorescent tag, O-2-[aminoethyl]fluorescein, which we have previously reported and shown to be suitable for the electrophoretic separation of carbohydrates in corn syrup samples. 47 This reagent has a high molar absorptivity similar to that of fluorescein and a good quantum yield and was shown to be suitable for the separation of seven corn syrup oligosaccharides.…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoretic separation of mono- and di-saccharides with dynamic pH junction and implementation in microchips

Kazarian

Hilder

Breadmore

2010

Analyst

View full text Add to dashboard Cite

An electrophoretic method for the separation of derivatised mono- and di-saccharides with on-line concentration via dynamic pH junction has been developed and optimised in capillaries. Dynamic pH junction is perfectly suited for on-line concentration of derivatised sugars due to the acidic derivatisation conditions, however, most reagents for carbohydrates are not ionisable, requiring the use of the novel reagent, O-2-[aminoethyl]fluorescein. Optimisation of the separation selectivity yielded best separations with 170 mM ammonium borate buffer at pH 8.60 in an acrylamide coated capillary. When using an injection comprising 7% of the capillary volume and detection via laser induced fluorescence (LIF) with an argon ion laser, limits of detection as low as 0.13 nM for maltose were obtained, which was 10 times lower than could be achieved without on-line concentration. In order to implement this system in a glass/PDMS microchip, the low pH sample was introduced into the microchannels via a cathodic pH independent electro-osmotic flow (EOF) generated using a poly(dimethyldiallylmethyl-ammonium chloride) (PDADMAC)/poly(styrene sulfonate) (PSS) polyelectrolyte multilayer coating. Optimisation of the injection volume in capillaries greatly simplified translation to the microchip platform, with the optimum capillary sample volume of 7%, dictating the use of an off-set cross with a volume 7% of the separation channel. Microchip separations of maltose, glucose, galactose and allose with dynamic pH junction, were achieved within 120 s, with the limit of detection of maltose using a light emitting diode induced fluorescence (LEDIF) detection system being 790 nM. This is 10 times lower than that achieved without concentration, and is lower than other reports of derivatised sugars using LEDIF detection. This is the first implementation of on-line concentration via a dynamic pH junction in a microchip, and significantly, the improvement in sensitivity achieved when translated to the microchip was equivalent to that achieved in capillaries.

show abstract

Development of a novel fluorescent tag O-2-[aminoethyl]fluorescein for the electrophoretic separation of oligosaccharides

Cited by 14 publications

References 27 publications

Online sample pre‐concentration via dynamic pH junction in capillary and microchip electrophoresis

Online sample pre‐concentration via dynamic pH junction in capillary and microchip electrophoresis

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2010–2012)

Capillary electrophoretic separation of mono- and di-saccharides with dynamic pH junction and implementation in microchips

Contact Info

Product

Resources

About