Vitamin C metabolomic mapping in experimental diabetes with 6-deoxy-6-fluoro-ascorbic acid and high resolution 19F-nuclear magnetic resonance spectroscopy.
Metabolomic mapping is an emerging discipline geared at providing information on a large number of metabolites as a complement to genomics and proteomics. Here we have probed ascorbic acid homeostasis and degradation in diabetes using 6-deoxy-6-fluoro ascorbic acid (F-ASA) and 750 MHz (19)F-nuclear magnetic resonance (NMR) spectroscopy with proton decoupling In vitro, Cu(2+)-mediated degradation of F-ASA revealed the formation of 4 major stable degradation products at 24 hours. However, when normal or diabetics rats were injected with F-ASA intraperitoneally (IP) for 4 days, up to 20 fluorine-labeled compounds were observed in the urine. Their composition resembled, in part, metal catalyzed degradation of F-ASA and was not explained by spontaneous degradation in the urine. Diabetes led to a dramatic increase in urinary F-ASA loss and a relative decrease in most other urinary F-compounds. Diabetes tilted F-ASA homeostasis toward oxidation in liver (P <.01), kidney (P <.01), spleen (P <.01), and plasma (P <.01), but tended to decrease oxidation in brain, adrenal glands, and heart. Surprisingly, however, besides the major oxidation product fluoro-dehydroascorbic acid (F-DHA), no F-ASA advanced catabolites were detected in tissues at 5 micromol/L sensitivity. These findings not only confirm the key role of the kidney in diabetes-mediated loss of ascorbic acid, but demonstrate that only selected tissues are prone to increased oxidation in diabetes. While the structure of most degradation products needs to be established, the method illustrates the power of high resolution (19)F-NMR spectroscopy for the mapping of complex metabolomic pathways in disease states.
Nishikawa
Yoko
Y
Institute of Pathology, Case Western Reserve University, Cleveland, OH 44106, USA.
Dmochowska
Barbara
B
Madaj
Janusz
J
Xue
Jie
J
Guo
Zhongwu
Z
Satake
Makoto
M
Reddy
D Venkat
DV
Rinaldi
Peter L
PL
Monnier
Vincent M
VM
D015083:2,3-Diketogulonic Acid / Q000138:chemical synthesis; D000818:Animals; D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism* / Q000652:urine; D001835:Body Weight; D003683:Dehydroascorbic Acid / Q000097:blood / Q000652:urine; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D005461:Fluorine; D005462:Fluorine Radioisotopes; D006706:Homeostasis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009929:Organ Size; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley
D005462:Fluorine Radioisotopes; C076401:6-deoxy-6-fluoroascorbic acid; D005461:Fluorine; D015083:2,3-Diketogulonic Acid; D001205:Ascorbic Acid; D003683:Dehydroascorbic Acid