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31 P NMR spectra of intact red blood cells: Quantitation of UDPGlucose and UDPGalactose
Author(s) -
Wehrli Suzanne L.,
Palmieri Michael J.,
Reynolds Robert A.,
Segal Stanton
Publication year - 1993
Publication title -
magnetic resonance in medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.696
H-Index - 225
eISSN - 1522-2594
pISSN - 0740-3194
DOI - 10.1002/mrm.1910300413
Subject(s) - chemistry , uridine , chromatography , quantitative analysis (chemistry) , nuclear magnetic resonance spectroscopy , erythrocyte membrane , nmr spectra database , hemoglobin , membrane , biochemistry , stereochemistry , spectral line , rna , physics , astronomy , gene
Abstract The combined levels of uridine diphosphogalactose (UDPGal) and uridine diphosphoglucose (UDPGlu) were measured directly by 31 P 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.