High-performance liquid chromatography of N,O-acylated sialic acids
β Scribed by Ashok K. Shukla; Norbert Scholz; Ernst H. Reimerdes; Roland Schauer
- Publisher
- Elsevier Science
- Year
- 1982
- Tongue
- English
- Weight
- 368 KB
- Volume
- 123
- Category
- Article
- ISSN
- 0003-2697
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An HPLC method has been developed for the assay of cytidine monophosphate-sialic acid synthetase (EC 2.7.7.43) using ion-pair chromatography and gradient elution. This procedure permits the assay of alternative substrates and inhibitors of the enzyme and is not subject to the limitation of the color
Sialic acids were specifically labeled with o-phenylenediamine-2HCl (OPD) to yield stable fluorescent quinoxaline derivatives. The sialic acids were released from the glycoprotein in a NaHSO4 solution (0.25 M 80 degrees C, 20 min) and derivatized in the same solution with the OPD (10 mg/ml (final co