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Capillary Electrophoresis of Carbohydrates pp 23–51Cite as

Fluorophores and Chromophores for the Separation of Carbohydrates by Capillary Electrophoresis

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Abstract

Carbohydrates are known to be the most abundant assembly of biopolymers with immense diversity, as a consequence of the complex glycosylation processes that occur within living organisms. This complexity inevitably complicates the ability to study these structures (Wells et al., Science 291:2376–2378, 2001). Viral infection and tumor malignancy have been pinpointed as a result of subtle changes in the structure of oligosaccharides, implying the significance of carbohydrates and ultimately the importance of the field of glycobiology (Chen et al., Nat Med 3:866–871, 1997; Isogai et al. Cancer Res 56:3902–3908, 1996). In most cases capillary electrophoresis (CE) offers greater advantages for studying carbohydrates when compared to other techniques, resulting in utilization of smaller samples, better separation efficiency, and shorter separation time. Typical analysis of carbohydrates in CE requires incorporation of a chromophore or a fluorophore through derivatization, which in due course enhances sensitivity and can also improve the separation of the analytes. This chapter discusses the currently used derivatization procedures, their mechanisms, and the advantages of each approach. The discussion mainly focuses on the publications from 2003 to 2009.

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Abbreviations

AcOH:

acetic acid

ANDSA:

7-aminonaphthalene-1,3-disulfonic acid

CBQCA:

3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde

CE:

capillary electrophoresis

CL:

chemiluminescence

DAD:

diode array detection

DMSO:

dimethylsulfoxide

HVAox:

2,2′-hydroxy-3,3′-dimethoxybiphenyl-5,5′-diacetic acid

Kdo:

2-keto-3-deoxyoctuolosonic acid

LED:

light-emitting diode

LIF:

laser-induced fluorescence

LODs:

limits of detection (measured in nM unless stated otherwise)

MALDI:

matrix-assisted laser desorption ionisation

MeOH:

methanol

NBD-F:

7-nitro-2,1,3-benzoxadiazole 4-fluoride

NMP:

1-(2-naphthyl)-3-methyl-5-pyrazolone

NPBA:

3-nitrophenylboronic acid

NS:

not stated

PEG:

polyethylene glycol

PITC:

phenylisothiocyanate

PMP:

1-phenyl-3-methyl-2-pyrazolin-5-one

QY:

quantum yield

SA:

sulfanilic acid

SAP:

(S)-3-amino-1,2-propanediol

SDS:

sodium dodecyl sulfate

THF:

tetrahydrofuran

TRSE:

4-carboxytetramethylrhodamine succinimidyl ester

UV:

Ultra violet

Vis (or vis):

visible

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Authors and Affiliations

  1. School of Chemistry, University of Tasmania, 75, Hobart Tas, 7001, Australia

    Michael Breadmore

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  1. Michael Breadmore
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  2. Emily Hilder
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  3. Artaches Kazarian
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Correspondence to Michael Breadmore .

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  1. Dipto. Biologia, Università di Modena e Reggio Emilia, Via Campi 213/D, Modena, 41100, Italy

    Nicola Volpi

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Breadmore, M., Hilder, E., Kazarian, A. (2011). Fluorophores and Chromophores for the Separation of Carbohydrates by Capillary Electrophoresis. In: Volpi, N. (eds) Capillary Electrophoresis of Carbohydrates. Humana Press. https://doi.org/10.1007/978-1-60761-875-1_2

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