1996
DOI: 10.1007/s0021663560339
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HPLC analysis of carbohydrates on POLYSPHER®CH OH columns using pulsed amperometric detection (PAD) with sodium hydroxide as post column detection reagent

Abstract: Carbohydrates have been separated on POLYSPHER(R)CH OH columns using pulsed amperometric detection (PAD) and UV detection (lambda =196 nm) in series and pure water as mobile phase. Nearly baseline separations have been obtained for the glycoprotein carbohydrates of sialic acid ( N-acetylneuraminic acid, NANA), N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc). As carbohydrates dissolved and eluted with pure water are present in the neutral form they are not detectable with PAD in contrast to carb… Show more

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Cited by 25 publications
(8 citation statements)
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“…Another disadvantage is that sialic acids cannot be determined in the same analysis with neutral and amino sugars. While high-performance anion-exchange chromatography with pulsed-amperometry detection (HPAEC-PAD) 2 (7,8) is easy, sensitive, and useful for the analysis of underivatized neutral and amino sugars liberated from glycoproteins, hydrolyzed amino acids and peptides cause significant interference (7).…”
mentioning
confidence: 99%
“…Another disadvantage is that sialic acids cannot be determined in the same analysis with neutral and amino sugars. While high-performance anion-exchange chromatography with pulsed-amperometry detection (HPAEC-PAD) 2 (7,8) is easy, sensitive, and useful for the analysis of underivatized neutral and amino sugars liberated from glycoproteins, hydrolyzed amino acids and peptides cause significant interference (7).…”
mentioning
confidence: 99%
“…With the exception of some minor peaks in the retention range below 6 min the phytosiderophores are the only analytes detectable by pulsed amperometric detection (PAD). This high selectivity and sensitivity of PAD for compounds containing aliphatic hydroxy groups is well-known and has been used mainly for detection of carbohydrates [15,16,17,18,19,20,21], although amino acids [22,23] and even metal species [24,25] are also detectable by PAD. The above mentioned peaks in the retention range below 6 min are amino acids, which are electroactive under the experimental conditions used.…”
Section: Resultsmentioning
confidence: 99%
“…Because this process is difficult to automate, it was performed manually. We chose sulfuric acid as hydrolyzing agent, because of the high recovery expected [12] and its compatibility with chromatographic separation on the Aminex column. Trifluoroacetic acid (TFA) is retained on the Aminex column, and is, therefore, less suitable than sulfuric acid.…”
Section: Hydrolysis Of Glycoproteinsmentioning
confidence: 99%