Use of reversed polarity and a pressure gradient in the analysis of disaccharide composition of heparin by capillary electrophoresis

Ruiz-Calero, Victoria; Puignou, L.; Galcerán, M.T.

doi:10.1016/s0021-9673(98)00662-1

Cited by 51 publications

(53 citation statements)

References 25 publications

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“…Heparin shows the highest sensitivity, followed by LMW heparin and heparan sulfate. Percentages of molar disaccharide compositions for unfractionated heparins from porcine, bovine intestinal mucosa and from bovine lung were determined by CE in a 60 mM formic acid buffer (pH 3.4; Ruiz-Calero et al, 1998). In addition, LMW heparin from bovine and porcine intestinal mucosa was analyzed as well (Ruiz-Calero et al, 1998).…”

Section: Ce Analysis Of Underivatized Gags Hyaluronanmentioning

confidence: 99%

“…The resolution of reversed-polarity separation decreases with increased oligosaccharide size (Pervin et al, 1994b). The eight heparin-derived disaccharides obtained by enzymatic depolymerization were separated by CE under a 60 mM formic acid buffer (pH 3.4), detection in the UV at 232 nm (Ruiz-Calero et al, 1998). The purity of 13 homogenous heparan sulfate-derived oligosaccharides depolymerized by heparin lyase II or heparin lyase III was demonstrated by the presence of a single peak on reversed polarity CE ( Fig.…”

Section: Ce Analysis Of Underivatized Gag-derived Oligosaccharidesmentioning

confidence: 99%

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Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan‐derived oligosaccharides

Mao

Thanawiroon

Linhardt

2002

Biomedical Chromatography

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Glycosaminoglycans are a family of polydisperse, highly sulfated complex mixtures of linear polysaccharides that are involved in many life processes. Defining the structure of glycosaminoglycans is an important factor in elucidating their structure-activity relationship. Capillary electrophoresis has emerged as a highly promising technique consuming an extremely small amount of sample and capable of rapid, high-resolution separation, characterization and quantitation of analytes. Numerous capillary electrophoresis methods for analysis of intact glycosaminoglycans and glycosaminoglycan-derived oligosaccharides have been developed. These methods allow for both qualitative and quantitative analysis with a high level of sensitivity. This review is concerned with separation methods of capillary electrophoresis, detection methods and applications to several aspects of research into glycosaminoglycans and glycosaminoglycan-derived oligosaccharides. The importance of capillary electrophoresis in biological and pharmaceutical samples in glycobiology and carbohydrate biochemistry and its possible applications in disease diagnosis and monitoring chemical synthesis are described.

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Section: Ce Analysis Of Underivatized Gags Hyaluronanmentioning

confidence: 99%

Section: Ce Analysis Of Underivatized Gag-derived Oligosaccharidesmentioning

confidence: 99%

Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan‐derived oligosaccharides

Mao

Thanawiroon

Linhardt

2002

Biomedical Chromatography

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“…Due to the high charge density and size of these molecules, it is the most documented method, allowing separation according to mass and charge. However, most of the reported work involves the analysis of oligosaccharides of small degree of polymerization such as the disaccharides generated by enzymatic degradations (Linhardt and Pervin, 1996;Karamanos et al, 1996;Toida and Linhardt, 1996;Grimshaw, 1997;Ruiz-Calero et al, 1998;Damm et al, 1992;Pervin et al, 1994;Desai et al, 1993). Only a few applications of capillary electrophoresis to intact anionic polysaccharides have been reported (Malsch et al, 1995(Malsch et al, , 1996Grimshaw et al, 1994;Roberts et al, 1998;Hayase et al, 1997;Mala et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Analysis of intact heparin by capillary electrophoresis using short end injection configuration

Duchemin¹,

Potier

Troubat³

et al. 2002

Biomedical Chromatography

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A capillary electrophoresis method for the analysis of intact heparin was developed using a phosphate buffer and a fused silica capillary. Operational parameters such as pH and concentration of the running buffer were investigated. The short end injection configuration permitted a gain on peak efficiency, on the analysis time and on the repeatability of both migration times and peak areas, through a reduction of the migration distance. Moreover, the beneficial effect of the presence of sodium chloride in the heparin sample on the peak efficiency was demonstrated and the influence of the salts on the conformation of the heparin was discussed. The optimized method (short end injection configuration, 50mM phosphate buffer pH 3, heparin sample prepared in 10 g/L NaCl solution) was validated in terms of linearity, reproducibility and specificity according to ICH requirements.

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“…In the last few years, capillary electrophoresis (CE) has proven to be a very attractive alternative separation technique for heparin oligosaccharides because it affords high resolving power and great flexibility in the separation orders. [17][18][19] CE is anticipated to provide oligomeric separation over a wider mass range than LC. It also enables faster separations and requires smaller sample quantities.…”

mentioning

confidence: 99%

“…The CE separation of heparin oligosaccharides has already been described in the literature. [17][18][19] These authors reported high efficient separations and investigated the influence of various parameters. These separations were, however, achieved with mobile phases not compatible with MS.…”

mentioning

confidence: 99%

Identification of heparin oligosaccharides by direct coupling of capillary electrophoresis/Ionspray‐Mass spectrometry

Duteil

Gareil

Girault³

et al. 1999

Rapid Commun. Mass Spectrom.

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A direct coupling between capillary electrophoresis (CE) and ionspray (IS) mass spectrometry (MS) has been optimized to identify oligosaccharides obtained by enzymatic digestion of heparin. The separation electrolyte was made compatible with the requirements of a direct coupling using electrolytes made of ammonium acetate buffer, pH 3.5 and 9.2. The different parameters of the CE/IS interface were optimized using a standard mixture of disaccharides: flow rate and composition of the sheath liquid, flow rate of the sheath gas and position of the capillary in the needle. Different combinations of positive or negative CE voltage polarity and positive or negative MS ionization modes were investigated. They allowed for detection orders to be easily reversed and for complementary structural information to be gathered. Finally, the optimized methodology was applied to the separation and characterization of porcine mucosa heparin depolymerized by heparinases II and III.

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Use of reversed polarity and a pressure gradient in the analysis of disaccharide composition of heparin by capillary electrophoresis

Cited by 51 publications

References 25 publications

Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan‐derived oligosaccharides

Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan‐derived oligosaccharides

Analysis of intact heparin by capillary electrophoresis using short end injection configuration

Identification of heparin oligosaccharides by direct coupling of capillary electrophoresis/Ionspray‐Mass spectrometry

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