Detection of carbohydrates using new labeling reagent 1-(2-naphthyl)-3-methyl-5-pyrazolone by capillary zone electrophoresis with absorbance (UV).

A novel labeling reagent 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) coupled with capillary electrophoresis (CE) with DAD detection for the determination of carbohydrates has been developed. The chromophore in the 1-phenyl-3-methyl-5-pyrazolone (PMP) reagent is replaced by naphthyl functional group, which results in a reagent with very high molar absorptivity (epsilon251 nm = 5.58 x 10(4) L mol(-1) cm(-1)). This permits NMP-labeled carbohydrates to be detected with UV absorbance in standard 50-mum-i.d. fused silica capillaries by zone electrophoresis. In this mode, nanomolar concentrations of detection limits are obtained. The method for the derivatization of carbohydrates with NMP is simplified. The derivatization reaction is rapid and mild in the presence of ammonia catalyst without further transfer steps. Nine monosaccharide derivatives such as mannose, galacturonic acid, glucuronic acid, rhamnose, glucose, galactose, xylose, arabinose and fucose can successfully be detected in CE mode. Good reproducibility can be obtained with relative standard deviation (R.S.D.) values of the migration times and peak area, respectively, from 0.44 to 0.48 and from 3.2 to 4.8. Furthermore, the developed method has been successfully applied to the analysis of carbohydrates in the hydrolyzed rape bee pollen samples.

[1]  J. Kennedy,et al.  Analysis of phenyldimethylsilyl derivatives of monosaccharides and their role in high-performance liquid chromatography of carbohydrates , 1983 .

[2]  M. Okamoto,et al.  Sensitive detection and structural characterization of trimethyl(p-aminophenyl)-ammonium-derivatized oligosaccharides by electrospray ionization-mass spectrometry and tandem mass spectrometry. , 1995, Rapid communications in mass spectrometry : RCM.

[3]  L. Zhang,et al.  Determination of 1-phenyl-3-methyl-5-pyrazolone-labeled carbohydrates by liquid chromatography and micellar electrokinetic chromatography. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[4]  J. Henion,et al.  Characterization of N-linked oligosaccharides by electrospray and tandem mass spectrometry. , 1992, Analytical chemistry.

[5]  M. Novotny,et al.  Capillary zone electrophoresis of oligosaccharides derivatized with N-(4-aminobenzoyl)-L-glutamic acid for ultraviolet absorbance detection. , 1997, Journal of chromatography. A.

[6]  J. Chmelík,et al.  Determination of neutral oligosaccharides in vegetables by matrix-assisted laser desorption/ionization mass spectrometry , 2004 .

[7]  S. Hase,et al.  Analysis of pyridylaminated O-linked sugar chains by two-dimensional sugar mapping. , 1996, Analytical biochemistry.

[8]  S. Honda,et al.  Determination of hyaluronic acid by high-performance liquid chromatography of the oligosaccharides derived therefrom as 1-(4-methoxy)phenyl-3-methyl-5-pyrazolone derivatives. , 1993, Journal of chromatography.

[9]  C. Sweeley,et al.  Structural characterization of oligosaccharides by high-performance liquid chromatography, fast-atom bombardment-mass spectrometry, and exoglycosidase digestion. , 1984, Analytical biochemistry.

[10]  S. Honda,et al.  Comparison of the sensitivities of various derivatives of oligosaccharides in LC/MS with fast atom bombardment and electrospray ionization interfaces. , 1996, Analytical chemistry.

[11]  Y. Lee,et al.  Precolumn labeling of reducing carbohydrates with 1-(p-methoxy)phenyl-3-methyl-5-pyrazolone: analysis of neutral and sialic acid-containing oligosaccharides found in glycoproteins. , 1991, Analytical biochemistry.

[12]  O. Dahlman,et al.  Efficient capillary zone electrophoretic separation of wood-derived neutral and acidic mono- and oligosaccharides. , 1996, Journal of chromatography. A.

[13]  K. Yoshino,et al.  Sensitive Analysis of Oligosaccharides Derivatized with 4-Aminobenzoic Acid 2-(Diethylamino)ethyl Ester by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry , 1996 .

[14]  J. Kennedy,et al.  Analysis of derivatives of carbohydrates by high-pressure liquid chromatography , 1979 .

[15]  K. Nakanishi,et al.  Characterization of methyl glycosides at the pico- to nano-gram level , 1983 .

[16]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[17]  J. Langridge,et al.  High-performance liquid chromatographic analysis of complex N-linked glycans derivatized with 2-aminoacridone. , 1997, Analytical chemistry.

[18]  P. Camilleri,et al.  High resolution and rapid analysis of branched oligosaccharides by capillary electrophoresis. , 1995, Analytical biochemistry.

[19]  R. O'neill,et al.  Monosaccharide composition analysis of oligosaccharides and glycoproteins by high-performance liquid chromatography. , 1995, Analytical biochemistry.

[20]  H. Perreault,et al.  Characterization of carbohydrates using a combination of derivatization, high-performance liquid chromatography and mass spectrometry. , 1998, Journal of chromatography. A.

[21]  S Hase,et al.  Reexamination of the pyridylamination used for fluorescence labeling of oligosaccharides and its application to glycoproteins. , 1984, Journal of biochemistry.

[22]  A. Kondo,et al.  Improved method for fluorescence labeling of sugar chains with sialic acid residues. , 1990, Agricultural and biological chemistry.

[23]  R. Linhardt,et al.  Analysis of fluorescently labeled sugars by reversed-phase ion-pairing high-performance liquid chromatography. , 1995, Journal of chromatographic science.

[24]  D. J. Strydom,et al.  Chromatographic separation of 1-phenyl-3-methyl-5-pyrazolone-derivatized neutral, acidic and basic aldoses , 1994 .

[25]  J. Nakamura,et al.  High-performance liquid chromatography of reducing carbohydrates as strongly ultraviolet-absorbing and electrochemically sensitive 1-phenyl-3-methyl-5-pyrazolone derivatives. , 1989, Analytical biochemistry.

[26]  C. Bush,et al.  High-performance liquid chromatographic separation of amino sugars and peptides with metal ion-modified mobile phases , 1982 .

[27]  A. Taga,et al.  Unusual separation of 1 -phenyl-3-methyl-5-pyrazolone derivatives of aldoses by capillary zone electrophoresis , 1997 .

[28]  K. Mopper,et al.  Reversed-phase liquid chromatographic analysis of dns-sugars : Optimization of derivatization and chromatographic procedures and applications to natural samples , 1983 .