31P NMR spectra of intact red blood cells: Quantitation of UDPGlucose and UDPGalactose

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Abstract
The combined levels of uridine diphosphogalactose (UDPGal) and uridine diphosphoglucose (UDPGlu) were measured directly by 31P NMR spectroscopy in intact fully oxygenated erythrocytes. Quantitative analysis was obtained using a sealed capillary of methylene diphosphonate (MDP) calibrated with standard solutions of UDPGlu and UDPGal of known concentration prepared in KRB/BSA. The combined peaks of UDPGlu and UDPGal were integrated after subtraction of the underlying broad component originating from membrane phospholipids. The average concentration of 27.16 ± 5.18 nmole/ml or 8.08 ± 1.36 μmole/100 g hemoglobin obtained for these metabolites correlated well with their total determined by HPLC of trichloracetic acid (TCA) extracts of the same samples.