Simultaneous analysis of a series of phosphorylated sugars in small tissue samples by anion exchange chromatography and pulsed amperometric detection

✍ Scribed by Steef M. de Bruijn; Richard G. F. Visser; Dr. Dick Vreugdenhil

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 79 KB
• Volume: 10
• Category: Article
• ISSN: 0958-0344
• DOI: 10.1002/(sici)1099-1565(199905/06)10:3<107::aid-pca449>3.0.co;2-l

✦ Synopsis

A simple method was developed to analyse a series of phosphorylated sugars in small physiological samples with dry tissue weights as low as 10 mg. Phosphorylated compounds were extracted from the tissue after boiling in 80% methanol by re-suspension of the dried methanol phase in water. The sugar phosphates were separated by Dionex high performance liquid chromatography and detected by pulsed amperometry. In contrast to the common protocol for the separation of sugars, the sugar phosphates were eluted at a low temperature and at a relatively low concentration of hydroxide in order to preserve the stability of the nucleotide sugars ADP-glucose and UDP-glucose. At low concentrations, the separation, detection and recovery of these compounds was highly satisfactory and an almost linear relationship between concentration and response was obtained. Quantities as low as 0.1 nmol could easily be detected.