USE OF [P-32) POLYPHOSPHORIC ACID AS A PHOSPHORYLATING AGENT - SYNTHESES OF P-32 LABELED SUGAR PHOSPHATES

Phosphorylation of D‐fructose with [32P]polyphosphoric acid followed by prolonged mild acid hydrolysis at 35 °C gave [32P]D‐fructose 1, 6‐diphosphate. The diphosphate was purified by conversion to the acid‐barium salt. [32P]D‐fructose 6‐phosphate was prepared by two methods : Degradation of the diphosphate by heating with 1N HBr for 1 hour at 88‐90 °C, or, replacing the prolonged mild acid hydrolysis of the phosphorylation mixture in the case of the diphosphate with a one hour treatment at 88‐90 °C. [32P]D‐glucose 6‐phosphate was prepared by dissolving the barium salt of [32P]D‐fructose 6‐phosphate in water and adding a dialyzed solution of phosphoglucose isomerase. The highly insoluble heptahydrate of the barium salt of [32P]D‐glucose 6‐phosphate readily crystallized from solution. Phosphorylation of D‐mannose with [32P]polyphosphoric acid gave [32P]D‐mannose 6‐phosphate, which upon reduction with KBH4, yielded [32P]D‐mannitol 1‐phosphate. Copyright © 1969 John Wiley & Sons, Ltd.