P-31 NMR-SPECTRA OF INTACT RED-BLOOD-CELLS - QUANTITATION OF UDPGLUCOSE AND UDPGALACTOSE

The combined levels of uridine diphosphogalactose (UDPGal) and uridine diphosphoglucose (UDPGlu) were measured directly by P-31 NMR spectros copy in intact fully oxygenated erythrocytes. Quantitative analysis wa s obtained using a sealed capillary of methylene diphosphonate (MDP) c alibrated with standard solutions of UDPGlu and UDPGal of known concen tration prepared in KRB/BSA. The combined peaks of UDPGlu and UDPGal w ere integrated after subtraction of the underlying broad component ori ginating from membrane phospholipids. The average concentration of 27. 16 +/- 5.18 nmole/ml or 8.08 +/- 1.36 mumole/100 g hemoglobin obtained for these metabolites correlated well with their total determined by HPLC of trichloracetic acid (TCA) extracts of the same samples.