Separation of ten phosphorylated mono-and disaccharides using HILIC and ion-pairing interactions.
Anal Chim Acta
; 972: 102-110, 2017 Jun 15.
Article
en En
| MEDLINE
| ID: mdl-28495090
Phosphorylated carbohydrates are indispensable cogs in several key metabolic wheels for all forms of life. Here, a straightforward liquid chromatography method coupled to mass spectrometry detection was developed for phosphorylated sugars. For separation of the targeted compounds, hydrophilic interaction chromatography (HILIC) was used with a bridged-ethylene hybrid amide column under alkaline conditions using triethylamine as a mobile phase modifier. Methylphosphonic acid was added to the aqueous mobile phase to reduce the tailing of compounds containing phosphate groups, which are known to interact with stainless steel components of the separation system. Under alkaline conditions and addition of methylphosphonic acid, the retention behavior can be attributed to both conventional HILIC mechanisms as well as ion-pairing interactions in the mobile phase. This hypothesis is supported by comparing the retention behavior of phosphorylated sugars and unmodified sugars. The HILIC method resolved eight biologically important phosphorylated sugars and thereby enables simultaneous detection and quantification of these compounds: fructose-1,6-bisphosphate, glucose-1-phosphate, glucose-6-phosphate, lactose-1-phosphate, mannose-6-phosphate, ribose-5-phosphate, sucrose-6-phosphate, and threhalose-6-phosphate. Fructose-1-phosphate and fructose-6-phosphate were not resolved but quantification of total fructose-phosphate is possible.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Espectrometría de Masas
/
Carbohidratos
/
Cromatografía Liquida
/
Disacáridos
Idioma:
En
Revista:
Anal Chim Acta
Año:
2017
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Países Bajos