Separation of 8-aminonapthalene-1,3,6-trisulfonic acid-labelled neutral and sialylated N-linked complex oligosaccharides by capillary electrophoresis

Klockow, Antje; Amadò, Renato; Widmer, Hans; Paulus, Aran

doi:10.1016/0021-9673(95)00597-g

Cited by 55 publications

(29 citation statements)

References 32 publications

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“…From these reasons, the present derivatization reaction could be considered to proceed quantitatively. The detection limits values were almost same or lower than those given by previously reported CE-LIF analyses of maltose using ANTS (50 nM) [29] and APTS (0.5 nM) [22]. Therefore, the present method is highly sensitive for saccharide analysis.…”

Section: Derivatization Conditionssupporting

confidence: 50%

“…A variety of reagents have been used for CE-LIF detection including APTS [22][23][24][25][26], 3-aminobenzoic acid [27,28], and 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS) [29,30]. However, the absorption maximum wavelengths of the above-mentioned reagents are not perfectly suited to the Ar ion and the He/Cd laser emissions at 488 and 325 nm, respectively.…”

Section: Introductionmentioning

confidence: 98%

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Highly sensitive capillary electrophoresis analysis of N‐linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser‐induced fluorescence detection

et al. 2011

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We describe a highly sensitive CE with laser-induced fluorescence (LIF) detection for the analysis of N-linked oligosaccharides in glycoproteins using rhodamine 110 as a fluorescence derivatization reagent. One CE separation is performed using a fused-silica capillary and neutral pH buffer conditions and allows for the separation of sialo-oligosaccharides according to the number of sialic acids. An alternate separation is performed using the same capillary and acidic pH buffer conditions, enabling the separation of asialo-oligosaccharides according to their sizes. The derivatization and separation conditions for the analysis of sialo- and asialo-oligosaccharides were optimized. Furthermore, we applied the proposed method for the analyses of N-linked sialo- and asialo-oligosaccharides in glycoproteins (ribonuclease B, fetuin, and recombinant human erythropoietin).

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Section: Derivatization Conditionssupporting

confidence: 50%

Section: Introductionmentioning

confidence: 98%

Highly sensitive capillary electrophoresis analysis of N‐linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser‐induced fluorescence detection

et al. 2011

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“…Precolumn labelling is instrumentally simpler and gives higher yields of derivatives compared to post-column reactions. Although lower detection limits can be reached by fluorescence-tagging procedures [1,2], UV detectors are by far the most usual ones and, on the other hand, differences between both tagging techniques can be considerably reduced by adequate selection of reagents.…”

Section: Introductionmentioning

confidence: 98%

Precolumn derivatization of reducing carbohydrates with 4-(3-Methyl-5-oxo-2-pyrazolin-1-yl) benzoic acid. Study of reaction, high-performance liquid chromatographic separation and quantitative performance of method

2002

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SummaryA simple method for quantitative determination of carbohydrates by reversed-phase, highperformance liquid chromatography after pre-column clerivatization and UV detection has been developed. Reducing sugars are condensed with 4-(3-methyl-5-oxo-2-pyrazohn-Lyl) benzoic acid (PMPA) to yield UVaclducts absorbing at 271 nm, which are resolved under typical reversed-phase conditions. After clerivatization, excess PMPA is easily removed from the reaction mixture by precipitation with mineral acids at pH < 4. The influence of experimental conditions on reaction yield, as well as chromatographic separation of derivatives, were investigatecl. The quantitative performancewas evaluatd by means of a protocol comprising replicate measurements at several analyte levels. The calibration curves obtained for 8 sugars showed excellent hnearity over 10 -5000 pmol. Limits of detection and quantification for several monosaccharicles were ca. 10 and 50 picomoles, respectively. Optimized conditions were successfully used for quantitative determination of monosaccharicles released after hydrolysis from fetuin, mucin, ~l-acid glycoprotein, ovalbumin and transferrin.

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“…CE evolved as a promising alternative in carbohydrate analysis with respect to fast and highly efficient separations [38]. Commonly carbohydrates are analyzed underivatized, while derivatization of oligosaccharides enables high-resolution CE [34].…”

Section: Introductionmentioning

confidence: 99%

CE‐MSⁿ of complex pectin‐derived oligomers

Coenen¹,

Kabel²,

Schols³

et al. 2008

Electrophoresis

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As pectin molecules are too large and heterogeneous to analyze as a whole, the polymer is usually degraded to smaller oligomers, which are often analyzed by high-performance anion exchange chromatography (HPAEC). However, the high salt concentration necessary to elute pectin oligomers by HPAEC is incompatible with online mass detection. To overcome such a disadvantage, a CE-IT-MS system was set up to further elucidate the fine structure of charged oligosaccharides. An effective separation of differently substituted galacturonic acid containing oligomers was obtained by low-pH CE-LIF analysis. By adapting the buffer and capillary online MS detection was enabled. Moreover, with MS/MS it was possible to localize sugar residues' substitutions. With this combined CE-MS approach LIF electropherograms of xylogalacturonan and rhamnogalacturonan I digests could be annotated. The method was further exemplified by a complex oligomer mixture of acid hydrolyzed apple pectin, which was separated and characterized by CE-MSn. Oligomers present in low amounts could be localized by their corresponding m/z, as was demonstrated by selected mass range representation.

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Separation of 8-aminonapthalene-1,3,6-trisulfonic acid-labelled neutral and sialylated N-linked complex oligosaccharides by capillary electrophoresis

Cited by 55 publications

References 32 publications

Highly sensitive capillary electrophoresis analysis of N‐linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser‐induced fluorescence detection

Highly sensitive capillary electrophoresis analysis of N‐linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser‐induced fluorescence detection

Precolumn derivatization of reducing carbohydrates with 4-(3-Methyl-5-oxo-2-pyrazolin-1-yl) benzoic acid. Study of reaction, high-performance liquid chromatographic separation and quantitative performance of method

CE‐MSⁿ of complex pectin‐derived oligomers

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Separation of 8-aminonapthalene-1,3,6-trisulfonic acid-labelled neutral and sialylated N-linked complex oligosaccharides by capillary electrophoresis

Cited by 55 publications

References 32 publications

Highly sensitive capillary electrophoresis analysis of N‐linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser‐induced fluorescence detection

Highly sensitive capillary electrophoresis analysis of N‐linked oligosaccharides in glycoproteins following fluorescence derivatization with rhodamine 110 and laser‐induced fluorescence detection

Precolumn derivatization of reducing carbohydrates with 4-(3-Methyl-5-oxo-2-pyrazolin-1-yl) benzoic acid. Study of reaction, high-performance liquid chromatographic separation and quantitative performance of method

CE‐MSn of complex pectin‐derived oligomers

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CE‐MSⁿ of complex pectin‐derived oligomers