HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SYSTEM FOR SEPARATING SUGAR PHOSPHATES A

2020-04-05 17:04:55

strength mm ionic anion intermediates

责任者: SWEZEY, RR 单位: STANFORD UNIV,DEPT BIOL SCI,HOPKINS MARINE STN,PACIFIC GROVE,CA 93950. 来源出处: JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS, v 669, JUL 21 1995, p 171- 176 摘要: A simple method for separating intermediates of carbohydrate metabolism, including the difficult-to-resolve sugar phosphates, using anion-exchange high-performance liquid chromatography is described. A gradient of decreasing berate concentration (10 to 0 mM) and increasing ionic strength (0 to 150 mM NH4Cl) separates phosphorylated sugars based on the strength of the ester complex that they form with berate anion. Lyophillized samples are reconstituted in a low ionic strength aqueous medium (5 mM triethanolamine-HCl, pH 8.1) and chromatographed on a commercially available anion-exchange column (Hamilton PRP-X100). The process requires only 3 h and permits nanomole detection of the phosphorylated intermediates of the glycolytic and pentose shunt pathways. 关键词: ASSAY METHODS; CHITIN; CORNEA; FUNGAL KERATITIS (KERATOMYCOSIS); CELL