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An Hplc and Epr Investigation On the Stability of Dmpo and Dmpo Spin Adducts in Vivo

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Application of the spin trapping technique in intact animals requires an understanding of the stability and distribution of the spin traps and their spin adducts in vivo. We studied the stability of DMPO in vivo in mice using HPLC and the stability of spin adducts of DMPO by EPR in plasma, whole blood, peritoneal fluid, and homogenized heart tissue of the rat. At 15 minutes after intraperitoneal injection DMPO had similar concentrations in the liver, heart, and blood of the mice and 40% remained in the organs 2 hours after the injection. In contrast, the spin adduct DMPO-OH was short lived, with a half-life of 3.0 minutes in plasma, and was not detectable 1 minute after formation in whole blood and homogenized heart tissue. The carbon centered spin adduct DMPO-CH(OH)CH3 was more stable, having half-lives of 16, 11, 3.6, and 0.79 minutes in plasma, peritoneal fluid, whole blood, and homogenized heart tissue, respectively. The spin adduct DMPO-SO3 was sufficiently stable for the adduct to be observed directly from living mice.

Affiliations: 1: Laboratory of Molecular Biophysics, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, NC 27709 USA; 2: Department of Radiology, Dartmouth Medical School, Hanover, NH 03755; 3: Biodynamic Laboratories, University of Wisconsin, Madison, WI 53706; 4: Laboratory of Experimental Pathology, National Cancer Institute, Bethesda, MD 20892 USA


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